CN113406318A

CN113406318A - Device and controller for testing biological samples

Info

Publication number: CN113406318A
Application number: CN202110783910.1A
Authority: CN
Inventors: 徐振腾; 张志宾; 黄光立; 戚玉桥; 张家伟; 王炯翰
Original assignee: Bonraybio Co Ltd
Current assignee: Bonraybio Co Ltd
Priority date: 2019-06-17
Filing date: 2020-06-17
Publication date: 2021-09-17
Also published as: TW202136849A; TWI755755B; CN112098405A; TWI808435B; TW202107148A

Abstract

The invention provides a device and a controller for testing a biological sample. The apparatus includes a receiving mechanism to receive the carrier. The carrier includes a holding area. The apparatus may include a camera module configured to capture a plurality of images of a holding area including a first image and a second image. The apparatus includes a positioning mechanism operable to adjust the relative position of the carrier with respect to the camera module. A processor in the device may identify an edge of the first image with the camera, cause the positioning mechanism to adjust the relative position of the carrier to the camera module such that when the camera acquires the second image, the edge of the second image is aligned with the identified edge of the first image, and perform a set of analysis processing routines on the image combined from the first and second images.

Description

Device and controller for testing biological samples

This application is a divisional application of the patent application entitled "automatic test apparatus for improving analytical accuracy" filed on 17.06/2020, application No. 202010554604.6.

Technical Field

The present invention relates to an apparatus for testing biological samples, and more particularly, to a testing apparatus having a magnifying function or an analyte-quantifying function.

Background

At present, testing of liquid contents is often entrusted to specialized testing facilities, using expensive microscope equipment with high magnification ratios to perform the testing. Since the ordinary person does not have a microscope device, the ordinary person cannot perform the test activity.

However, in some of today's test categories it is necessary to perform periodic tests; thus, the need for frequent testing is overburdened in terms of time and expense. For example, a category of long-term testing includes the semen testing of infertile patients. The semen test is mainly aimed at observing the number of sperm, their motility and morphology.

The semen testing method comprises the following steps: semen from a male subject is allowed to stand at room temperature for a period of time, a drop of the sample is taken, the sample is instilled onto a glass slide, and the sample is observed under a microscope. These observations include not only highly magnified observations of individual sperm to identify the external topography of individual sperm, but also observations of large numbers of whole sperm, their motility, morphology and number per unit area. However, individuals cannot perform semen tests themselves, as the industry has not developed techniques that allow individuals to perform tests via simple accessory devices.

Disclosure of Invention

The present disclosure includes methods, devices and systems for testing biological samples. In one example aspect, an apparatus for testing a biological sample is disclosed. The device includes a receiving mechanism to receive a carrier including a holding area configured to carry a biological sample.

The device includes a camera module configured to capture an aggregate image (image) of the holding area and a processor configured to identify visual cues on a carrier from the captured aggregate image of the holding area using the camera module and to perform a set of analysis processing routines on the captured aggregate image based on the identification of the visual cues. The aforementioned identification of the visual cue further comprises verifying whether the holding area carries the biological sample based on the manner in which the visual cue is displayed in the captured collective image, and selectively causing the processor to perform the analysis processing routine on the captured collective image according to the result of the verification. In another example aspect, a device for testing a biological sample includes a receiving mechanism to receive a carrier including a holding area configured to carry or have been exposed to the biological sample. The apparatus includes a camera module configured to capture a plurality of images of a holding area, and a processor configured to adaptively select an analysis algorithm suitable for a movement characteristic of a biological sample under test using the camera module based on the plurality of images of the holding area and to execute a set of analysis processing routines corresponding to the analysis algorithm selected by capturing the plurality of images to generate an analysis result regarding the biological sample.

In another example aspect, a device for testing a biological sample includes a receiving mechanism to receive a carrier including a holding area configured to carry or have been exposed to the biological sample. The device includes a camera configured to capture a plurality of images of a holding area including a first image, a second image. The apparatus also includes a positioning mechanism operable to adjust the relative position of the carrier to the camera and a processor configured to identify edges of the first image with the camera module, cause the positioning mechanism to adjust the relative position of the carrier to the camera such that when the camera acquires the second image, the edges of the second image are aligned with the identified edges of the first image, and perform a set of analytical processing routines on the image combined from the first and second images to determine one or more characteristics of the biological sample.

In another example aspect, an apparatus for testing a biological sample includes a receiving mechanism to receive a carrier including a holding area, wherein the holding area carries or has been exposed to the biological sample. The apparatus includes a camera module configured to capture a collective image of the holding area. The camera module further includes a focus motor operable to adjust a focus of the camera. The apparatus also includes a processor configured to determine volumetric characteristics of the holding area based on operation of the focus motor with the camera module and perform a set of analytical processing routines on at least a portion of the captured set of images of the holding area to determine one or more characteristics of the biological sample. One or more characteristic receiving processors of the biological sample receive an adjustment based on the determined volumetric characteristic of the holding area.

These and other features of the present technology will be described in this document.

Drawings

Fig. 1A is an exploded view of an apparatus for testing a biological sample according to an embodiment of the present invention.

FIG. 1B is an assembled view of the testing device of FIG. 1A.

FIG. 2A is a cross-sectional view of the test device of FIG. 1A.

FIG. 2B is a cross-sectional view of another embodiment of a testing device.

FIG. 3 is a flow chart of the testing device according to an embodiment of the present invention.

Fig. 4 is a cross-sectional view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 5 is a cross-sectional view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 6 is a schematic view of the test apparatus of fig. 5 in use.

Fig. 7 is a schematic view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 8 is a schematic view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 9 is a schematic view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 10 is a schematic view of an apparatus for testing a biological sample according to another embodiment of the present invention.

Fig. 11 to 13 are views of apparatuses for testing biological samples according to another three embodiments of the present invention.

FIG. 14A is a schematic view of a test strip inserted into a meter device according to another embodiment of the present invention.

Fig. 14B is a schematic diagram of elements of a meter device according to another embodiment of the present invention.

Figure 15A shows a sample processing routine for semen testing by a device such as a meter device or smart communication device.

Fig. 15B shows a sample step 1515 of the processing routine shown in fig. 15A.

Fig. 15C shows a sample step 1520 of the processing routine shown in fig. 15A.

Fig. 15D shows a sample step 1530 of the processing routine shown in fig. 15A.

FIG. 15E shows sample step 1550 of the processing routine shown in FIG. 15A.

Fig. 15F shows sample step 1555 of the processing routine shown in fig. 15A.

Fig. 16 shows a sample processing routine for determining sperm cell concentration.

Fig. 17 shows sample sperm and sample sperm trajectories.

Fig. 18 shows a sample processing routine for determining sperm cell trajectory and motility.

Fig. 19 is a schematic view of a testing device including a collection bottle.

Fig. 20 is a schematic view of a testing device that does not include a collection bottle.

Fig. 21A, 21B, and 21C are schematic cross-sectional views of various embodiments of a test apparatus.

FIG. 22 is a cross-sectional schematic view of a test strip device having two sample holding areas.

Fig. 23 is a schematic diagram of elements of a test apparatus having an auto-focus function.

FIG. 24 is a schematic diagram of the components of another test apparatus with an auto-focus function.

Fig. 25A and 25B are schematic cross-sectional views of a testing apparatus including a switch and a motor.

Fig. 26A and 26B are schematic cross-sectional views of a testing device including a flexible element.

FIG. 27 is a flow chart of a processing routine for analyzing a semen sample of a male client or patient.

FIG. 28 is a flow chart of a process routine for analyzing LH or HCG in a female client or patient.

Fig. 29 shows an example of a carrier that may be suitable for a testing device having a multi-camera setup, such as the testing device shown in fig. 22.

Fig. 30 is a flow chart of a processing routine for analyzing fertility of both male and female subjects utilizing the testing device disclosed herein.

Fig. 31 shows an additional example of an analytical processing routine with a visual cue (e.g., in or near the holding area) that the carrier may be used to perform with the control test device.

FIG. 32 is an additional example flow diagram of a processing routine that may be implemented by the disclosed testing device to adaptively perform analytical processing routines based on visual cues.

FIG. 33 is an example flow diagram of a processing routine that may be implemented by the disclosed test apparatus.

Fig. 34 is an example image of a holding area divided into a number of blocks.

Fig. 35 is a sample image illustrating a portion of a candidate block selection process routine.

FIG. 36 is an example image illustrating the results after an image processing program (e.g., two polarization) and cell number determination.

FIG. 37 is a flow chart of an example calibration routine that may be implemented by the disclosed test apparatus.

FIG. 38 test carrier carries an example image of a visual cue and/or image pattern that may be used to correct or validate a test device.

FIG. 39 is an example image of the visual cue of FIG. 38 captured by the disclosed testing device.

Fig. 40A and 40B illustrate different image qualities of different blocks in the captured image of fig. 39.

FIG. 41 is an image of a test carrier carrying an example test sample that can be used to calibrate or validate a test device.

Fig. 42A and 42B illustrate different image qualities of different blocks in the captured image of fig. 41.

FIG. 43 is a flow diagram of an example of an implementable processing routine for the testing apparatus disclosed herein to verify that the sample-holding region carries a biological sample.

Fig. 44A shows an example of an captured image of an empty or dry sample-holding area.

Fig. 44B shows another example of an captured image of an empty or dry sample-holding area.

Fig. 45A shows an example of a sample holding area loaded with an captured image of a fluid carrying a biological sample.

Fig. 45B shows another example of a sample-holding region loaded with an captured image of a fluid carrying a biological sample.

FIG. 46 illustrates an example image including a number of misclassified regions.

FIG. 47 is a flow diagram of an example of an implementable processing routine for determining movement characteristics of a sample by the testing device of the present invention.

FIG. 48 is a flow diagram of an example of an implementable processing routine for utilizing multiple fields of view by the test equipment disclosed herein to improve results.

Fig. 49 shows an exemplary multi-axis mobile platform (multi-axis mobile platform).

Fig. 50A shows an example manner in which images are sequentially acquired in a clock sequence as viewed from a camera.

Fig. 50B shows an example manner of acquiring images in a sequential scanning (progressive scan) order.

Fig. 51A illustrates an example of the preset positions of the adjustable lenses of the camera module.

FIG. 51B shows the camera module of FIG. 51A with the lens in an extended position.

Fig. 52 shows an example of a configuration for determining the actual volume of the sample contained in the carrier.

Detailed Description

Fig. 1A and 1B illustrate an apparatus for testing a biological sample according to an embodiment of the present invention. The specific examples disclosed herein are for illustrative purposes and are not to be construed as required limitations of the present invention. The apparatus a1 for testing a biological sample comprises: a carrier 10 having a sample holding area 11 formed on top of the carrier 10, a cover 20 stacked on top of the carrier 10, and at least one magnifying member 30 (also referred to as a magnifying element or magnifying lens) comprising a convex lens-type surface formed on the cover 20.

The enlargement part 30 of the present specific example includes a planar convex lens as shown in fig. 1A. However, other types of magnifying lenses (e.g., double-sided lenticular lenses) may be included as the magnifying member 30. The amplifying member 30 is disposed to align with the sample holding area 11 of the carrier 10 and cover the sample holding area 11. The amplification part 30 may have various amplification ratios based on test requirements of various tests. For example, the test may include a semen test, a urine test, a synovial joint fluid test, a skin test, a water test, or other bodily fluid test, among others.

The test using the device for testing a biological sample a1 of the present embodiment does not require an additional magnifying lens or a laboratory microscope, which is expensive and time-consuming to operate. Furthermore, there is no need to align the sample holding area with the magnifying lens or a laboratory microscope.

As shown in fig. 1A, the sample-holding region 11 of the carrier 10 may be formed with a recessed structure. The recessed structure design provides a stable and large storage space containing the sample 40. The recessed structure allows the sample to rest for a desired period of time before performing a test. For example, before performing motility tests on semen samples, it is necessary to leave the semen sample at room temperature for a desired period of time before performing the motility test.

The sample 40 may first be dripped into the recessed structure (i.e., the sample holding area 11 of the carrier 10) to rest for a period of time. As shown in fig. 1B, the total area of the cover 20 may be less than the total area of the carrier 10. A sample receiving port 12 exposed to the outside of the housing 20 is formed on one side of the sample holding area 11. The sample receiving port 12 may be designed to have an outwardly expanding shape that may help to smoothly drip a sample.

Fig. 2A shows air channels 13 extending beyond the other side of the housing 20 and formed on the other side of the sample holding area 11. The air channel 13 may prevent air from filling the interior of the sample holding area 11, which would prevent receipt of the sample when it is in a liquid state.

As shown in fig. 2A, the side illumination device 50 may be disposed at a side of the carrier 10 of the test device a 1. The side illumination device 50 may provide illumination to the sample 40 in the sample holding area 11 and thus improve the resolution of the captured test image of the sample 40. In some embodiments, the sample holding area 11 may receive illumination from a light source on the top or bottom of the testing device a 1.

As shown in fig. 1A, the enlarged member 30 and the housing 20 may be integrally formed, i.e., the enlarged member 30 and the housing 20 may be a single element. In other embodiments, such as the embodiment shown in fig. 2B, the removable housing 20 and the enlargement 30 disposed in the upwardly-facing concave member, i.e., the recess 21, of the removable housing 20 may each be separate components adapted to be integrated together. In other words, the same type of removable housing 20 may be integrated with different magnification components 30 at various magnification ratios.

In some embodiments, the distance between the bottom of the detachable cover 20 and the sample holding area 11 is 0.005mm to 10 mm. In some embodiments, the distance between the bottom of the detachable cover 20 and the sample holding area 11 is about 0.01 mm. The testing device may comprise one or more spacers (not shown) to ensure the distance between the bottom of the detachable cover 20 and the sample holding area 11. The spacer may be integrally formed with the removable cover 20 or the sample holding area 11 of the carrier 10.

In some embodiments, the strip comprising the carrier 10 and the cover 20 is used for sperm testing. In some embodiments, the optimal angular magnification ratio for determining sperm concentration and motility is about 100 to 200. In some embodiments, the optimal angular magnification ratio for determining sperm morphology is about 200 to 300. The thinner the magnification element, the higher the angular magnification ratio.

The focal length of the magnifying element may also be related to the angular magnification ratio. In some embodiments, a magnifying element having an angular magnification ratio of 100 has a focal length of 2.19 mm. The magnifying element with an angular magnification ratio of 156 has a focal length of 1.61 mm. The magnifying element having an angular magnification ratio of 300 has a focal length of 0.73 mm. In some embodiments, the magnifying element has an angular magnification ratio of at least 30, preferably at least 50. In some embodiments, the focal length of the magnifying element is 0.1mm to 3 mm.

Fig. 3 shows a sample processing routine for performing a test using the apparatus for testing a biological sample a1 shown in fig. 1B. In step S110, a sample 40 to be tested is disposed in the sample holding area 11. In step S110, the cover 20 is stacked on top of the carrier 10, and then the sample 40 to be tested is disposed in the sample holding area 11 from the sample receiving port 12. Alternatively, the sample 40 to be tested may first be placed directly in the sample holding area 11, after which the cover 20 is stacked on top of the carrier 10. In step S120, the sample 40 is rested in the sample holding area 11, optionally for a period of time, according to the test requirements of the sample 40. In step S130, an intelligent communication device (e.g., a mobile phone) is attached to the housing 20, and a camera of the mobile phone is aligned with the magnifying part 30 to capture an image or video data of the sample via the magnifying part 30 using the camera of the mobile phone. In step S140, application code (APP) running at the mobile phone or other analysis device may be used to perform analysis on the image or video material, thereby obtaining a test result.

As shown in fig. 4, a support side (such as a protruding member 14) may be further formed on the top of the housing 20 of the test device a2 at the boundary of the enlarged member 30. In some embodiments, a protrusion-type support structure may be formed on top of the housing 20 by adding protrusion members 14. When a user attempts to capture image or video data of a sample using the smart communication device 60 (e.g., a mobile device such as a smart phone or a single board computer), the side of the smart communication device 60 having the camera 61 may be secured to the protruding member 14 (in the direction shown by arrow L1). Thus, test device A2 allows a user to capture image or video data of a sample using the intelligent communications device 60 without the need for expensive test devices to record the image or video data. Further, for the optimum observation distance, the height of the protruding member 14 may be predetermined based on the specifications of the camera 61 and the testing device a 2.

As shown in fig. 5 and 6, test device a3 may include a meter device 70 (also referred to as a base assembly). The gauge device 70 includes a lower barrel base 71 and an upper barrel body 72 that can be raised or lowered relative to the lower barrel base 71. The lower barrel base 71 has an insertion port 73 that provides an insertion location for the stacked housing 20 and carrier 10. A light emitting device (also referred to as a light source) 80 facing upward is disposed on the bottom of the lower barrel base 71 to provide illumination from the bottom to the combination of the housing 20 and the carrier 10. The upper barrel body 72 may include, for example, at least one additional magnifying lens 74 for further magnification.

The upper barrel body 72 may be attached to the lower barrel base 71 using a threaded mechanism so that the upper barrel body 72 can be raised or lowered relative to the lower barrel body 71 as screws. In other words, the upper barrel body 72 is rotatable relative to the lower barrel base 71 in the direction of arrow L2 such that the upper barrel body 72 moves upward and downward relative to the lower barrel base 71 in the direction of arrow L3. By adjusting the height of the upper barrel body 72 relative to the lower barrel body 71, the system adjusts the height of the magnifying lens 74 (changes the magnification ratio) and the height of the camera 61.

An assembly frame 75 (also referred to as a form-fitting frame) may be disposed at an upper end of the upper barrel body 72. The assembly frame 75 fixes the intelligent communication device 60 at a predetermined position. The assembly frame 75 has a camera alignment hole 76. The camera 61 of the smart communication device 60 may receive light from the sample through the camera alignment aperture 76.

The camera 61 disposed on the current smart communication device 60 typically has only a digital zoom function. In general, high precision testing requires an optical zoom lens. However, the user using the test apparatus a3 did not need the camera 61 with an optical zoom lens. The height adjustment function of test device a3 provides a flexible solution for aligning the sample, magnifying lens, and camera 61.

Fig. 6 shows the smart communication device 60 assembled and secured to an assembly frame 75, the assembly frame 75 being disposed on the upper barrel body 72. The housing 20 and the carrier 10 containing the sample 40 are inserted through the insertion port 73. The upward facing light emitting device 80 can provide illumination to the sample and increase the brightness of the sample.

The upper barrel body 72 or the gauge device 70 may be rotated in the direction L2 to adjust the height of the magnifying lens 74 and the camera 61 up or down in the direction L3. The height adjusting mechanism realizes a function of adjusting the magnification ratio. The camera 61 may capture motion video data or still test images of the sample 40 after being magnified. In addition, the intelligent communication device 60 can use its original equipment functions to store captured video data or images, deliver test images or video data, and perform subsequent processing.

As shown in fig. 7, the apparatus for testing a biological sample a4 includes a plurality of magnifying

members

30, 30B, 30C having different magnification ratios disposed on the housing 20. The user may deflect the cover 20 to align the sample holding region 11 of the carrier 10 with any of the magnifying

members

30, 30B, 30C having different magnification ratios in order to obtain test results having different magnification ratios. By this design, the test apparatus a4 with a single module amplification function can be applied to satisfy the amplification requirements of multiple test protocols without changing the amplification part or the housing.

As shown in fig. 8, the device a5 for testing a biological sample includes a flexible transparent film 15. A flexible transparent film 15 is disposed between the carrier 10 and the amplifying member 30 and covers the sample holding area 11. The flexible transparent film 15 covers the specimen 40 (in a liquid state) so that the specimen 40 is in a confined space. Thus, external influences caused by air, dust and dirt are restrained to a minimum. In addition, the testing apparatus A5 can adjust the focus by changing the thickness of the flexible transparent film 15.

As shown in fig. 9, the magnifying part 30 of the apparatus for testing a biological sample a6 is a planar convex lens, and the surface of the magnifying part 30 facing the carrier 10 is a protruding surface. Thus, an upwardly-facing concave hollow part, i.e., a groove 21, is formed at the surface of the enlargement part 30 facing the carrier 10. The focal length parameter H1 is defined by the thickness of the thickest part of the enlargement portion 30 of the planar convex lens. As shown in fig. 10, the focal length parameter H2 of the apparatus for testing a biological sample a7 is different from the focal length parameter H1 of fig. 9.

The focal lengths H1 and H2 can be adjusted by changing the thickness of the housing 20 or the amount of curvature of the magnifying member 30. For example, the focal length H2 shown in fig. 10 is greater than the focal length H1 shown in fig. 9, and is achieved by changing the magnitude of the curvature of the magnifying member 30. In this way, various focal length test requirements can be met by employing different amplifying components 30.

In some embodiments, the magnifying member 30 may be transparent and the remainder of the housing 20 may be opaque. In addition, the carrier 10 may comprise a transparent sample holding area 11. The remainder of the carrier 10 may be opaque. When performing a test operation on a test device, light may propagate through the sample holding area 11, amplifying component 30 to suppress the chance of light interference in other components of the device.

Referring to fig. 11, in the apparatus for testing a biological sample A8, the carrier 10 of the testing apparatus A8 further includes a beam assist guide structure 16 formed at the bottom surface of the carrier 10. The carrier 10 may be made of a transparent or translucent material. The beam assist guide structure 16 may be opaque or include a granular structure, a roughened pattern, an engraved pattern, or other suitable structure that scatters the light beam reaching the guide structure 16. The beam assist guide structure 16 may provide a specific pattern to the entire surface or a part of the surface of the housing and the carrier. The beam assist guide structure 16 may also be formed entirely around the side surface of the carrier 10.

When the housing 20 and the carrier 10 are stacked and attached to the smart communication device 60 (e.g., as shown in fig. 4), the amplifying part 30 is aligned with the camera 61 of the smart communication device 60. Additionally, supplemental light (not shown) may be disposed on the surface of smart communication device 60 in the vicinity of camera 61. A light beam provided by the supplemental light may be directed to the carrier 10 to illuminate the sample holding area 11 through the cover 20. At the same time, the beam-assisted guiding structure 16 of the carrier 10 may scatter the light beam provided by the supplemental light, thereby improving the brightness and illumination uniformity of the sample-holding area 11.

By positioning the beam assist guide structure 16, the test apparatus does not require an additional supplemental light source to illuminate the carrier 10. Accordingly, the housing 20 includes light transmissive material so that supplemental light from the smart communication device 60 can pass through the housing 20 to reach the sample. In some alternative embodiments, the device does not include the cover 20 and the supplemental light reaches the carrier 10 directly without propagating through the cover 20.

The device for testing biological samples A8 may include a non-slip film 92, a release paper 96, and a pH test paper 94. The slip prevention film 92 is attached on a supporting side (such as a top side) of the housing 20, and serves to stably mount the housing 20 to the camera 61 of the smart communicator 60, as shown in fig. 4, such that the amplifying part 30 is aligned with the camera 61 of the smart communicator 60. The positioning of the smart communicator 60 relative to the test device A8 is fixed to a predetermined configuration using the non-slip film 92.

The anti-slip film 92 may have an opening aligned with the magnifying member 30 such that the anti-slip film 92 does not block light transmitted from the sample to the camera 61 through the magnifying member 30. The non-slip film 92 may include a material such as silicon, and the surface of the non-slip film 92 may be protected with a release paper 96 to maintain the paper tack. A pH test paper 94 may be disposed on the sample holding area 11 of the carrier 10 to provide an indication of the pH value of the sample. The pH paper 94 may be replaced after use.

In addition, the enlargement part 30 and the housing 20 may be detachably designed. Therefore, the user can select another amplifying part 31 different from the amplifying part 30 to replace the original amplifying part 30 based on the test requirement. Various magnification components that may be assembled with the housing 20 are assembled to achieve different magnification ratios or other optical characteristics.

Referring now to fig. 12, a device for testing a biological sample a9 may further comprise a sample collection sheet 42 disposed in the sample holding area 11. The sample collection tab 42, for example, has a sample collection area 42A. The specimen collection area 42A may use adhesive or other methods to collect sperm, subcutaneous tissue/cells, parasite eggs, and similar solid test subjects. In some embodiments, the sample collection sheet 42 may serve as a spacer that maintains the distance between the housing 20 and the sample holding area 11.

Then, referring to fig. 13, a device for testing a biological sample a10 may include a spacer element 98 disposed at the sample holding area 11 between the carrier 10 and the enclosure 20. The isolation element 98 may isolate the amplifying member 30 from the test fluid in the sample holding area 11 and prevent the test fluid from contaminating the amplifying member 30. In some embodiments, spacer element 98 may serve as a spacer to maintain the distance between housing 20 and sample holding area 11. The isolation member 98 may be integrated with the housing 20 as a single component. Alternatively, the isolation component 98 may be integrated with the carrier 10 as a single component.

FIG. 14A is a schematic view of a test strip inserted into a meter device according to another embodiment of the present invention. The test strip 5 (also referred to as a test cartridge) includes a removable housing 20 and a carrier 10. In other words, the combination of the removable cover 20 and the carrier 10 (e.g., as shown in fig. 1B) forms the test strip 5. The test strip 5 is inserted into a meter device 70 (also referred to as a base assembly) via an insertion port. The insertion port may be, for example, a lateral or vertical insertion port. The meter device 70 may comprise, for example, means for capturing images of the sample collected in the test strip 5.

Fig. 14B is a schematic diagram of elements of a meter device according to another embodiment of the present invention. The meter device 70 includes an insertion port 73 that provides an insertion location for the test strip 5. The test strip 5 includes a carrier 10 and a removable cover 20. The removable housing includes an enlargement 30. The meter device 70 includes a camera 61 for capturing images or video data of the sample holding area of the carrier 10. The camera 61 is aligned with the magnifying member 30. The meter device further comprises a light source 80 for providing illumination to the sample holding area from the bottom. In some embodiments, a collimator (e.g., a collimator lens or light reflector; now shown) may be placed on top of the light source 80 for collimating the light beam. An annular aperture may further be placed between the light source 80 and the collimator so that the light beam traveling through the collimator forms a hollow cone shaped light beam. The carrier 10 may comprise a transparent or translucent material for light propagation.

In some embodiments, the meter device 70 may further include a phase plate that shifts the phase of the light emitted from the sample-holding region. As the light propagates through the sample, the velocity of the light increases or decreases. Thus, the light rays propagating through the sample are out of phase (about 90 degrees) with the remaining light rays that did not propagate through the sample. The out-of-phase light rays interfere with each other and enhance the contrast between the bright and dark portions of the sample image.

The phase plate may further shift the phase of the light propagating through the sample by about 90 degrees in order to further enhance the contrast caused by the interference of out-of-phase light. Thus, the light rays propagating through the sample are about 180 degrees out of phase with the remaining light rays that do not propagate through the sample. This destructive interference between light enhances the contrast of the sample image by darkening objects in the image and lightening their boundaries.

In some alternative embodiments, such a phase plate may be disposed on top of the detachable housing 20 of the test strip 5. In other words, the phase plate may be part of the test strip 5, not part of the meter device 70.

Fig. 15A-15F illustrate sample processing routines for semen testing by a device such as the meter device 70 or smart communication device 60 shown in fig. 5, 14A and 14B, respectively. In step 1505, the device obtains an image (frame) of the sample. In step 1510, the apparatus determines a sperm concentration based on the image. In step 1515, the device can further determine the pH of the sample by analyzing the color or grayscale of the pH band. For example, the device may include a processor to identify the color of the portion of the image captured by the camera (corresponding to the pH band) and determine the biochemical characteristics (e.g., pH level) of the biological sample contained in the band. In some other embodiments, the light source of the device can provide illumination having at least one color. For example, the light source may include light emitters having different colors (e.g., red, green, and blue) to form light of various colors. The camera of the apparatus may further capture at least one (or more) image(s) of the sample illuminated with the light. The processor may compare the color of a particular region of the image (e.g., a pH banding region) to determine a characteristic of the biological sample or a quantification of the analyte. In some embodiments, the processor only needs the color of a particular region of one image to determine the characteristic of the biological sample. For example, a device (e.g., a test device) may include a color correction module for correcting colors of an image. The processor then analyzes the corrected image to determine a characteristic of the biological sample. Alternatively, the test strip may include a color correction region having a known color. The processor performs a color correction operation on the image based on the color correction region, and then analyzes the corrected image to determine a characteristic of the biological sample or a quantification of the analyte. In some embodiments, the reagent in the pH strip (or other type of biochemical test strip) reacts with the biological sample, and a particular region of the post-image (e.g., a pH strip region) shows a particular color. In some embodiments, the specific area for color detection necessarily requires magnification of the image captured by the camera. Thus, at least in some embodiments, there is no amplification component or supplement above a particular region of the band used for color detection (e.g., the pH band region). For example, some types of biochemical test strips contain photochemical agents. When the photochemical reagent reacts with a specific analyte in the biological sample, the reaction causes a color change in the sample holding area of the strip. The processor may analyze images of the test strip (captured by the camera) to detect color changes and quantify specific analytes in the biological sample. In addition, the device can determine sperm morphology (1520), sperm volume (1525), and total number of sperm (1530). In step 1540, the device obtains a series of multiple frames of the sample. In

steps

1545, 1550 and 1555, the device can determine a sperm motility parameter and determine sperm motility based on the sperm cell trajectory. Fig. 16 shows a sample processing routine for determining sperm cell concentration. In 1605, a camera of the meter device 70 or the intelligent communication device 60 ("the device") as shown in fig. 5, 14A and 14B, respectively, captures magnified images of the sperm sample. The captured image is the original image used to determine sperm concentration. The device then converts the digital color image to a digital grayscale image, and further divides the digital grayscale image into a plurality of regions.

In step 1610, the device performs a self-adjusting threshold binary computation for each region based on the mean and standard deviation of its grayscale values. The goal of the self-adjusting bounding bin calculation is to identify objects that are candidates for sperm as foreground objects and the rest of the region as background.

Foreground objects in the image after the binary computation may still include impurities that are not actually sperm. Those impurities are smaller or larger than the sperm cells. The method can set an upper boundary value and a lower boundary value for the size of the sperm. In step 1615, the device performs a noise reduction operation on the image by removing impurities that are greater than the upper boundary value of the sperm or less than the lower boundary value of the sperm. After the denoising operation, the foreground objects in the image represent the sperm.

The method counts the number of sperm in the image based on the head portion of the sperm. In

steps

1620 and 1625, the device performs a distance transform operation to calculate a minimum distance between the foreground object and the background, and also identifies the location of the local maximum. That position is a candidate for sperm head position.

In step 1630, the device performs an ellipse fitting operation on each sperm candidate object to reduce false positive candidates that do not have an elliptical shape and are therefore not sperm heads. The device then counts the total number of remaining positive candidates for sperm and calculates the concentration of the sperm based on the volume represented by the image. The volume may be, for example, the area of the extracted sample-holding region multiplied by the distance between the sample-holding region and the bottom of the housing.

In some embodiments, the device may use multiple images of the sample and calculate the density values based on the images, respectively. Then, the apparatus calculates an average value of the concentration values so as to minimize a measurement error of the sperm concentration.

Using a series of images of the sample (e.g., a frame of video data), the device can further determine the trajectory and motility of the sperm. For example, fig. 17 shows sample sperm such as sperm 1705 and sample sperm trajectories such as trajectory 1710 and trajectory 1720.

Fig. 18 shows a sample processing routine for determining sperm cell trajectory and motility. A camera, such as the meter device 70 or the smart communication device 60 ("the device") shown in fig. 5, 14A, and 14B, respectively, captures a series of images (e.g., frames of video data) of a sperm sample. The device uses a series of images captured to determine a parameter of sperm motility. To determine the parameters of sperm motility, the device needs to track the trajectory of each sperm in the series of images.

The device converts a digital color image to a digital grayscale image. The device first identifies the head position of the sperm in the first image of the series (e.g., using the method shown in fig. 16). The identified head position of the sperm in the first image is the initial position of the sperm track to be tracked. In some embodiments, the device may use two-dimensional Kalman filtering (Kalman filter) to estimate the trajectory of the sperm's motion.

In some embodiments, the tracking has a measurement value z_j(k) Sperm of (S)_jThe two-dimensional Kalman filtering of (1) comprises the steps of:

1: computing predicted states

And error covariant matrix

2: using predicted states

Measured value z_j(k) And error covariant matrix

Calculating the predicted measurement values

Residual error of measured value

And residual covariant matrix

3: if it is

And

then the kalman filter gain is calculated

Updated state estimate

And an updated error covariant matrix

Representing the prediction of image k based on image k-1,

the position and speed of the jth sperm.

Q (k-1) is a co-variate matrix of the processing noise, N (k) is a co-variate matrix of the white position noise vector, γ is a gate threshold and V is a co-variate matrix of the estimation error_maxThe maximum possible sperm velocity.

The method can use joint probability data correlation when tracking multiple trajectories of multiple spermAnd (4) jointly filtering to determine a track path. The joint probability data correlation filter determines feasible joint correlation events between the detection target and the metrology target. Feasible federated Association events (A)_js) To determine the relative probability value between sperm s and sperm j. Next, the method makes a path allocation decision based on the best assignment method. A. the_jsIs defined as:

the lambda is a parameter which is the number of,

is a gaussian probability density function of the detected sperm.

Based on the series of frames over a period of time, the method identifies a trajectory for each sperm, such as trajectory 1805 as shown in fig. 18. The method then determines various parameters of sperm motility based on the trajectories. The parameters include: such as curve Velocity (VCL), straight-line Velocity (VSL), Linearity (LIN), and amplitude of lateral head displacement (ALH). The curve speed (VCL)1810 is defined as the sum of the moving distances in unit time. A linear Velocity (VSL)1815 is defined as a linear movement distance in a unit time. The Linearity (LIN) is defined as VSL divided by VCL. The lateral head displacement Amplitude (ALH)1820 is defined as twice the amplitude of the lateral displacement of the sperm head relative to the mean path 1825.

In some embodiments, the curve Velocity (VCL)1810 can be used to determine sperm motility. The method may set a speed threshold. Any sperm having a VCL above or equal to the velocity threshold are identified as motile. The remaining sperm having a VCL below the velocity threshold are identified as non-motile sperm. The level of motility is the number of identified motile sperm divided by the total number of sperm identified from the image.

The method can further analyze sperm morphology. The camera of the meter device 70 or the smart communication device 60 ("the device") captures magnified images of the sperm sample. The captured image is the original image for determining the morphology of the sperm.

The method detects the shape of a sperm candidate based on segmentation. The method uses the position of the sperm heads as an initiation point. The image of the sperm is segmented into a head portion, a neck portion, and a tail portion using a shape-dependent segmentation algorithm. For example, the method may segment the sperm using a method such as an active contour model.

Based on the sections, the method calculates parameters (such as length and width) for the various sections. The classifier (such as a support vector machine, neural network, convolutional neural network, or AdaBoost algorithm) may be trained using a training data set that includes already labeled samples. After training, parameters of various parts of the sperm can be fed into the classifier to determine whether the sperm has the proper morphology. In some embodiments, the classifier can be used for other applications such as detecting characteristics of cells and microorganisms.

Furthermore, it has been found in some cases that due to unstable voltages, flickering light sources or other types of noise, several areas of the image in the image may be mistaken for the motion trajectory of the sperm. FIG. 46 illustrates an example image including a number of misclassified regions. In fig. 46, a small area 4601 is classified as a motion trajectory, which is static in nature. To reduce external noise, the processor may employ an analysis algorithm to determine sperm motility characteristics to minimize misclassification.

FIG. 47 is a flow diagram of an example of an implementable processing routine 4700 for accurately determining motion characteristics of a sample by the test equipment disclosed herein. Step 4710, for example: after insertion into the carrier cassette, the device may capture multiple images of the sample holding area of the carrier using the camera module.

The device of step 4720 may adaptively select an analysis algorithm suitable for the motion characteristics of the biological sample being tested based on the plurality of images. In some embodiments, the movement characteristic indicates that the biological sample is substantially static or dynamic. After determining that the biological sample is substantially static, a static algorithm may be selected to process the captured image. In one example, the static algorithm determines morphology (morphology), such as sperm acrosome (acrosome) and/or mid-section of sperm. In some embodiments, the static algorithm may also analyze a Sperm Chromatin Dispersion (SCD) stain image to determine normality of sperm DNA fragments. On the other hand, after the biological sample is determined to be substantially dynamic, a dynamic operation may be selected. In another example, a dynamic algorithm may be used to determine the trajectory of high motility sperm. Dynamic algorithms help determine a number of parameters related to sperm motility, such as: such as VCL, VSL, VAP, ALH, etc., as shown in fig. 18, to optimize computational efficiency.

To select the analysis algorithm at step 4720, according to some embodiments, the device may first select two images from the plurality of images. The device then compares the two images to determine the amount of change between the first and second images to determine which analysis algorithm is appropriate for use. The amount of change can be determined based on the rate of change of movement of the detectable target in the biological sample. For example, the detection device may compare the amount of change between the two images to a predefined threshold, indicating that the biological sample characteristic is substantially static or dynamic. If the variance is less than a threshold, the biological sample is deemed static and a static analysis algorithm is selected to process the captured image. On the other hand, if the variation is greater than or equal to the threshold, the biological sample is determined to be dynamic, and a dynamic analysis algorithm is selected for subsequent processing.

The plurality of images may include a series of images acquired in a short time. In some embodiments, 2 to 600 images may be acquired in 0.04 to 10 seconds (e.g., at a rate of 60 images per second). In another specific example, the rate may be 15 images per second. For example, 45 images may be acquired in 3 seconds. The device may select two images that are each taken separately in time and/or order, thereby making the amount of change between the two images more noticeable. For example, the two images may be separated by 2 to 5 seconds (e.g., the first image and the last image in the sequence are selected).

In this manner, the device may perform a set of analysis processing routines corresponding to the selected analysis algorithm on the captured aggregate image to obtain an analysis result associated with the biological sample at step 4730.

Fig. 19 is a schematic view of a testing device including a collection bottle according to at least one embodiment of the present disclosure. The test strip tape device 1905 may be inserted into the test device 1900 through an insertion port. The test strip tape device 1905 can include a collection vial 1910 for collecting a sample (e.g., a sperm sample) or include a slot for receiving a collection vial. The testing device 1900 may include a sensor (not shown) to detect whether a collection vial 1910 is inserted into the testing device 1900.

Testing device 1900 may have a timer mechanism for determining the period of time over which collection vial 1910 is inserted into testing device 1900. After insertion of the collection vial 1910 containing the sample, the testing device 1900 may wait a predetermined period of time (e.g., 30 minutes) to liquefy the sample before prompting the user to transfer the sample from the collection vial 1910 to the test strip tape device 1905. In some embodiments, testing device 1900 may include a camera or sensor to determine whether the sample has liquefied.

Further, the testing device may include a movement mechanism to apply a mechanical force to the collection vial 1910 in order to mix the sample in the collection vial 1910. For example, the movement mechanism may, for example, shake, vibrate, or rotate the collection vial 1910. In some other embodiments, the testing device may include a shaft to be inserted into the collection vial 1910 and to agitate the sample in the collection vial 1910.

The test device 1900 may optionally include a display (e.g., screen 1920) for displaying data. For example, screen 1920 may show instructions or prompts as to how to operate test apparatus 1900. Screen 1920 may also show the results of the test after the test performed by test apparatus 1900. Additionally or alternatively, the test device 1900 may include known communication modules such that it can communicate (e.g., analyze results and/or images acquired by a camera module) with a user's computing device (e.g., a smart phone with mobile software application code, or a traditional personal computer such as a laptop). The test device 1900 may be operable to receive instructions from a user (e.g., from the screen 1920 and/or from the communication module) and execute a selected number of automated analysis processing routines based on the instructions. The test device 1900 may also display the results and/or images of the sample on the screen 1920 or on the user's computer (e.g., via the communication module described above), or both.

Similar to the test device shown in fig. 14A and 14B, the test device 1900 may include a camera (not shown) for capturing images or video data of the test strip tape device 1905. The test device 1900 may further include a processor (not shown) for processing image or video material (e.g., via the processing routine shown in FIG. 16) for determining the test results.

In some embodiments, for example, the magnifying element 2110 is a magnifying lens. The magnification power of the magnification assembly 2110 may be represented by an angular magnification ratio or a linear magnification ratio. The angular magnification ratio is the ratio between the angular size of the object as seen through the optical system and the angular size of the object as seen directly at the closest apparent distance (i.e., 250mm from the human eye). The linear magnification ratio is a ratio between the size of an image of an object to be projected on the image sensor and the size of the actual object.

For example, the magnifying lens may have a focal length of 6mm, a thickness of 1mm, and a diameter of 2 mm. Assuming that 250mm is the near point distance of the human eye (i.e., the closest distance the human eye can focus), the angular magnification ratio is 41.7 times 250 mm/6 mm. The distance between the amplification element 2110 and the sample holding region 2115 may be, for example, 9 mm. Therefore, the linear magnification ratio can be close to 2. In other words, the size of the image of the object on the image sensor caused by the magnifying element is 2 times the size of the actual object under the magnifying element.

In some embodiments, the magnifying element has a focal length of 0.1mm to 8.5 mm. In some embodiments, the linear magnification ratio of the magnifying element is at least 1. In some embodiments, the linear magnification ratio of the magnifying element is 0.5 to 10.0.

In some embodiments, a supplemental lens 2135 is placed below camera module 2130 for further magnifying the image and reducing the distance between magnifying element 2110 and sample holding area 2115. The effective linear magnification ratio of the entire optical system may be, for example, 3. In other words, the size of the image of the object captured by the camera module 2130 is 3 times the size of the actual object in the sample holding area 2115. In some embodiments, the effective linear magnification ratio of the entire optical system of the test apparatus is 1.0 to 100.0, preferably 1.0 to 48.0.

In some embodiments, the image sensor of the camera module has a primitive size of 1.4 μm. Typically, an acquired image of the object requires acquisition of at least 1 primitive in order to properly analyze the shape of the object. Therefore, the size of the captured image of the object needs to be at least 1.4 μm. If the linear magnification ratio of the test apparatus is 3, the test apparatus can properly analyze the shape of the object having a size of at least 0.47 μm.

In some embodiments, the image sensor of the camera module has a primitive size of 1.67 μm. Then, the size of the captured image of the object needs to be at least 1.67 μm in order to properly analyze the shape of the object. If the linear magnification ratio of the test apparatus is 3, the test apparatus can properly analyze the shape of the object having a size of at least 0.56 μm.

In some embodiments, the length of the entire optical system may be, for example, 24 mm. The distance between the bottom of the magnifying element and the top of the sample-holding region 2115 may be, for example, 1 mm. In some embodiments, the length of the entire optical system of the test device is 2mm to 100mm, preferably 5mm to 35 mm.

Fig. 20 is a schematic view of a testing device that does not include a collection bottle in accordance with at least one embodiment of the present disclosure. Unlike test device 1900, test device 2000 does not include a collection vial or a slot for insertion of a collection vial. The sample is applied directly to the test strip tape device 2005 by the user or operator without collection in a collection vial.

FIG. 21B is a schematic cross-sectional view A-A of the embodiment of the test apparatus 1900 of FIG. 21A. Section a-a of the test device 1900 shows a camera module 2130 on top of the test strip tape device 2105 for capturing images or video data of the sample holding area 2115 of the test strip tape device 2105. The test strip tape apparatus 2105 includes an amplification assembly 2110 on top of the sample holding area 2115. A light source 2140 below the test strip tape arrangement 2105 provides illumination to the sample holding area 2115. In some other embodiments, the light source may be placed on top of the test strip tape assembly or laterally on one side of the test strip tape assembly. There may be a plurality or array of light sources for providing illumination on the test strip tape means. In some embodiments, different combinations of light sources may be switched, adjusted, or selected depending on the analyte type so that the analyte is illuminated by light of the appropriate color.

In some embodiments, the test strip tape arrangement 2105 may include a test strip in or near the sample holding area 2115. For example, the test strip may be a pH test strip, a HCG (human chorionic gonadotropin) test strip, a LH (luteinizing hormone) test strip, or a fructose test strip. Some optical properties of the test strip (e.g., color or light intensity) may change when an analyte of a sample in the sample-holding region interacts with a chemical or biochemical reagent in the test strip. The camera module 2130 can capture the color or intensity of the test strip to determine the test result, such as pH level, HCG level, LH level or fructose level. In some embodiments, the magnifying element 2110 above the test strip may be replaced with a transparent or translucent cover. Thus, the test device may simultaneously check for an analyte in a sample and perform another analysis of the sample via one or more magnified images of the sample.

FIG. 21C is a schematic cross-sectional view A-A of another embodiment of the test apparatus 1900 of FIG. 21A. Section a-a of the testing device 1900 shows a camera module 2130 on top of a test strip tape device 2105 for capturing images or video data of a sample holding area 2115 of the test strip tape device 2105, which includes a sensor and one or more lenses 2135 (also referred to as supplemental lenses or optical lens modules). A light source 2140 below the test strip tape arrangement 2105 (or disposed elsewhere) provides illumination for the sample holding area 2115. The magnification component 2110 may be attached to the bottom of the lens 2135, rather than on the top of the sample holding area 2115 as shown in fig. 21A. In some embodiments, if the lens 2135 provides sufficient magnifying power, the element 2110 can be a flat light transmissive housing without magnifying power. In some other embodiments, if lens 2135 provides sufficient magnification (e.g., if the linear magnification ratio of lens 2135 is at least 1.0), then test device 1900 does not include magnification element 2110.

Fig. 22 is a schematic view of a test device with a test strip device having two sample holding areas. Fig. 29 shows an example of a carrier that may be suitable for a test apparatus having a multi-camera configuration, such as the test apparatus shown in fig. 22. Referring to fig. 19 and 20 together, the testing device shown in fig. 22 may be another variation of testing device 1900 (i.e., with a collection bottle) or testing device 2000 (i.e., without a collection bottle). As shown in fig. 22, a receiving mechanism is included in the testing device to receive one or more carriers (e.g., test strip tape apparatus such as test strip tape apparatus 2205, or a collection vial such as vial 1910) that are insertable through an opening on a housing of the testing device.

In some embodiments, a single carrier may include a first holding area and a second holding area, such as shown by test strip tape apparatus 2205 in fig. 22. As shown in fig. 22, at least two camera modules may be included in the test apparatus. The two camera modules include a first camera module 2230A and a second camera module 2230B configured to capture image and/or video data of the first holding area 2215A and the second holding area 2215B, respectively. More particularly, the test strip tape device 2205 may include a sample holding area 2215A and another sample holding area 2215B. In some examples, a transparent or translucent outer cover 2210A is placed on top of sample holding area 2215A. Light source 2240A may be controllable and may provide illumination on sample holding area 2215A. The first camera module 2230A is positioned to capture images or video data of the sample holding area 2215A. As an alternative implementation, the amplification element 2210B may be placed on top of the sample holding area 2215B. Additionally, in some embodiments, light source 2240B may be operable to provide illumination on sample holding area 2215B. The second camera module 2230B is positioned to capture images or video data of the sample-holding area 2215B. The first holding area and the second holding area may directly carry the biological sample or have been exposed to the biological sample. Similar to the structure introduced with respect to fig. 14B, in some embodiments, the testing device may include a collimator for collimating the light beam emitted by the light source to at least one of the holding areas. In some embodiments, an annular aperture may further be included between the light source and the collimator for forming a hollow cone-shaped beam of light that travels through the collimator and then to the sample holding area. In some additional embodiments, a phase plate may be included between the sample holding area and at least one of the camera modules for phase shifting light reflected by the sample holding area.

As an alternative to a single carrier having multiple holding areas, multiple carriers may be inserted into the test apparatus through their respective openings, ports, or slots. For example, two separate test strip devices may include

sample holding areas

2215A and 2215B, respectively. Depending on the needs of the test, the position of the

sample holding areas

2215A and 2215B in the test strip may be designed to align with the first camera module 2230A and the second camera module 2230B. In some embodiments, two test strip devices are inserted into the test device via two independent insertion ports.

The convenience and ease of testing are two significant benefits that the testing device disclosed herein can provide, among other benefits. According to embodiments herein, a user of the disclosed testing device does not have to possess any expertise on how to perform various types of analyses on biological samples before the user can utilize the testing device to produce results. Thus, the testing device may include a processor for performing automated analytical processing routines on the sample and determining results regarding the sample. The processor may be carried by a main circuit board (i.e., known components, not shown for simplicity). In addition, the testing device is preferably small and not as bulky as conventional testing devices commonly found in laboratories. Thus, in some embodiments, such as those shown in fig. 19 and 20, the receiving mechanism of the carrier, the camera module, and the main circuit board can all be packaged in the housing of the test deviceAnd (4) the following steps. The test device may have a small physical dimension, such as less than 30 centimeters (cm) by 30cm, i.e., 27,000cm³. In some embodiments, the testing device may further include a battery compartment enclosed within the housing such that a battery may be mounted in the battery compartment to power the testing device.

In some embodiments, a processor included in the test device may perform different analyses on different holding areas, and may compile results based on a combination of results of the analyses performed on the different areas. In other words, the processor may be configured to perform a first analysis processing routine on the captured image of the first holding area, perform a second analysis processing routine, different from the first analysis processing routine, on the captured image of the second holding area, and determine a result for the biological sample based on results of both the first analysis processing routine and the second analysis processing routine. As used herein, the term "analytical processing routine (analytical process)" means a processing routine that can evaluate one or more segments of data collected from a number of sources (e.g., images of a holding area) and produce results, conclusions, final results, estimates, or the like regarding the sources.

According to some examples, the testing device may use a combination of the first camera module 2230A, the light source 2240A, and the housing 2210A to quantify the analyte or determine a characteristic of the sample (e.g., a pH level, an LH level, an HCG level, or a fructose level). Additionally, the testing device may further analyze the magnified image of the sample using a combination of the second camera module 2230B, the light source 2240B, and the magnification element 2210B to determine characteristics of the sample (e.g., sperm count, sperm motility, sperm morphology, etc.). Depending on the requirements of various types of biochemical tests, different combinations or configurations of light sources may be used to illuminate the biochemical sample. The multiple camera configuration is particularly advantageous since different analytical processing routines can be performed via different camera modules without requiring the user to change the carrier (e.g., test strip tape device), thereby speeding up the generation of results and reducing the complexity of the necessary human operations.

Light sources

2240A and 2240B are enclosed inside the housing and are configured to illuminate the biological sample for at least one of the camera modules. According to one or more embodiments, the processor is configured to control the light source based on an analytical processing routine the processor is currently configured to perform.

Further, in some embodiments, the processor may perform different analysis processing routines based on the visual cues on the carrier. For example, some embodiments may perform image recognition and processing on an image of the holding area, and may perform different analysis processing routines according to visual cues from the results of the image recognition. Example vectors 2905(1) through 2905(4) are shown in fig. 29, where vector 2905(1) is used for fertility testing (e.g., via sperm thereof) on germ cells of a male subject, and vectors 2905(2), 2905(3), and 2905(4) are used for fertility testing (e.g., via urine thereof) on germ cells of a female subject. As shown, all of the carriers 2905(1) -2905 (4) have a first holding region 2915A corresponding to the position of the first camera module 2230A, but only the carrier 2905(1) includes a second holding region 2915B. In some examples, the visual cue on the carrier may be the shape of a particular holding area (e.g., holding area 2215A). For the discussion herein, the shape of the holding area means the overall edge (or outer perimeter) of the holding area. For example, the shape may be circular, oval, triangular, rectangular, or any suitable shape recognizable by a processor utilizing known image processing techniques on an image of the holding area as captured by a respective camera module (e.g., the first camera module 2230A). Additional examples of visual cues may include graphical patterns, visual signs, one-dimensional barcodes, multi-dimensional pattern codes (e.g., QR codes), and the like.

Referring to fig. 22 and 29 concurrently, in some embodiments, when the processor identifies (e.g., via the first camera module 2230A) that the first holding area (e.g., area 2215A or area 2915A of carrier 2905(1)) is in a first shape (e.g., circular), the processor is configured to perform a certain analysis processing routine (e.g., fertility of a male subject, such as according to various characteristics of its sperm sample), and when the area 2215A or area 2915A of the first holding area (carrier 2905(2)) is in a second shape (e.g., oval), the processor will perform a different analysis processing routine (e.g., analysis of fertility of a female subject, such as according to hormone level of its urine sample). In this way, the test device is not limited to performing only one type of test (e.g., sperm fertility), but it can also switch analytical processing routines accordingly based on the carrier inserted into the machine (e.g., test strip tape device).

More particularly, according to some implementations, when the shape indicates that the biological sample includes sperm from a male subject, the processing routine may determine one or more characteristics of the sperm, such as those introduced herein. In some examples, the determination of one or more characteristics of the sperm can be performed by using the second camera module 2230B. For some specific examples, characteristics that may be determined may include: sperm concentration, sperm motility, and/or sperm morphology. According to some embodiments, the processor is configured to (1) determine a concentration of sperm and/or a morphology of sperm based on a single one of the captured images, and (2) determine motility of sperm based on two or more of the captured images.

In view of the above, fig. 30 is a flow diagram of an example processing routine 3000 for analyzing fertility of both male and female subjects utilizing the testing device disclosed herein (e.g., in fig. 22). With continued reference to fig. 29, a process routine 3000 is described below. It should be noted that the following example applies a sample of a male first, followed by a sample of a female, but the reverse order (i.e., female and then male) may be performed without affecting the accuracy of the results.

First, in step 3002, a user may apply a biological sample (e.g., sperm) of a male subject to a first holding area (e.g., area 2915A) and a second holding area (area 2915B) of a first carrier (e.g., carrier 2905 (1)). Next, in step 3004, the user inserts the first carrier into the testing device (e.g., one shown in fig. 22), and because the shape of the first holding area 2915A of the carrier 2905(1) is circular, the testing device can automatically acquire knowledge that the current sample contains sperm from a male and select an analysis processing routine accordingly. Next, in step 3006, the user can determine one or more characteristics of the sperm using the testing device. For example, as discussed herein, a processor in the test device may utilize the first camera module 2230A to acquire an image of the first holding area 2915A of the carrier 2095(1) (the carrier 2095(1) may include test strips that show different colors in response to different acidity), and identify the color of the test strips to determine the acidity of the sperm. Additionally, in step 3006, the processor in the testing device may utilize the second camera module 2230B to determine one or more characteristics of sperm selected from the group consisting of: the number of cells (e.g., sperm count), the concentration of sperm, the motility of sperm, and the morphology of sperm.

Next, in step 3008, the user may apply urine from the female subject to the holding region 2915A of the second carrier (e.g., carrier 2905 (2)). In step 3010, the user inserts the second carrier into the testing device, and since the shape of the first holding region 2915A of the carrier 2905(2) is oval, the testing device may automatically acquire knowledge that the current sample contains urine from a woman and select an analytical processing routine accordingly. In step 3012, the testing device determines one or more characteristics of the urine, for example, by using the second camera module 2230B. For example, the test strip may be adapted to enable the test device to determine the concentration level of one or more types of female hormones (e.g., FSH, LH, or HCG). Finally, in step 3014, the user utilizes the testing device to automatically analyze the results of the male and female biological samples and determine a final result regarding the fertility of the subject.

In some particular examples, the first camera module 2230A may have a lower camera resolution than the second camera module 2230B, and thus the two cameras are used by the processor to perform different analysis processing routines. In addition, the first camera module 2230A may have a lower magnification ratio than the second camera module 2230B. Some examples of the first camera module 2230A may have no magnification function at all, while the second camera module 2230B may have a fixed magnification ratio. In addition or as an alternative to the second camera module 2230B having a higher magnification ratio by itself, the housing 2210B of the second holding area 2215B may include a magnification component, such as shown in fig. 22. In some implementations, the magnification ratio of the camera module may be adjustable (e.g., controlled by a processor). Some examples of testing devices provide that the first camera module 2230A has a camera resolution of 2 million primitives or greater and the second camera module 2230B has a camera resolution of 13 million primitives or greater. In some examples, the second camera module 2230B may include a linear magnification ratio of at least 4.8 times or more.

In some of these examples, the processor further determines at least one additional characteristic of the sperm by using the first camera module 2230A. This additional characteristic may include the acidity of the sperm. For example, the carrier may include a pH indicator that indicates the acidity of the sperm using a color in the first holding area 2215A that the processor may recognize to identify the acidity. Similarly, some examples provide that the processor may determine a biochemical characteristic of the biological sample based on a color of a region in one or more images of the first or second holding region.

Continuing with the above example of a testing device having a multi-camera configuration in fig. 22 and the example of a carrier in fig. 29, in some implementations, when the processor identifies that the first holding area (e.g., area 2215A or area 2915A of carrier 2905(2)) is in a second shape (such as an oval) that may indicate that the biological sample includes urine from a female subject, the processor is configured to determine one or more characteristics of the urine. Characteristics that may be determined may include: LH level, FSH level, and/or HCG level. As with the acidity, the determination of one or more characteristics of the urine can be performed by using the first camera module. Similarly, the carrier may include a LH indicator (e.g., as shown in carrier 2905 (3)), a FSH indicator (e.g., as shown in carrier 2905(2)), and/or a HCG indicator (e.g., as shown in carrier 2905 (4)) in a first holding area (e.g., area 2915A of the respective carrier).

Further, in some embodiments, the processor may utilize at least one of the two camera modules (e.g., the first camera module 2230A) or another sensor (e.g., the photosensor 2690 introduced below with respect to fig. 26B) to determine the readiness or validity of the biological sample prior to performing the analytical processing routine. In some implementations, the readiness or validity of the test sample can be determined based on identifying whether the first visual indicia is displayed in a particular region (e.g., where line 2916 is shown in fig. 29) of the first holding region (e.g., region 2915A). An example of such a first visual marker may be a line displayed in some designated area on the test strip, such as shown as red line 2916 in fig. 29. The red line 2916 may be used as a work quality control means, which may indicate that the test is valid or that the results are ready. Additionally, the first holding region 2915A may include another region that displays a second visual indicia representing a test result regarding a characteristic of the biological sample (e.g., where line 2917 is shown in fig. 29). An example of this second visual indicia may be a line displayed on the test strip in another certain designated area of readiness, such as shown as red line 2917 in fig. 29.

In some embodiments, the testing device may perform an action in response to determining that the biological sample is not ready. In some examples, the actions performed by the processor include implementing a timer having a duration determined by the analysis processing routine to be performed. In some other examples, the testing device further includes a movement mechanism, and a processor in the testing device may utilize the movement mechanism to apply a mechanical force to the carrier to improve the readiness of the biological sample. More details of actions and mechanisms that may be implemented in the test device are introduced below with respect to fig. 25A, 25B, 26A, and 26B.

The position of the magnifying elements (e.g., the magnifying element of the camera module or the magnifying component of the test strip) and the position of the light sources may be adjusted or selected as desired for various types of analyte analysis. In a variation, the camera modules may have adjustable magnification ratios. In at least some of these examples, the processor is further configured to adjust a magnification ratio of at least one of the two camera modules based on an analysis processing routine the processor is currently configured to perform. As introduced above, when the biological sample includes sperm, the testing device may configure a suitable camera module (e.g., the second camera module 2230B) to achieve different magnification ratios for determining sperm motility and sperm morphology.

It should be noted that the optimal distance between the camera module and the magnifying element may have a low margin of error. For example, even a small deviation of 0.01mm from the optimal distance may prevent the camera module from capturing a sharp image of the sample holding area. In order to finely adjust the distance between the camera module and the magnifying element, the testing device may include an Auto Focus (AF) function. An autofocus function is a function that automatically adjusts an optical system (e.g., adjusts the distance between elements of the optical system) so that an object being imaged (e.g., semen) is within the focal plane of the optical system. At least one or more embodiments also provide a mechanical focusing mechanism controllable by the processor to focus at least one of the two camera modules on a respective holding area. The mechanical focus mechanism is discussed in more detail below with respect to fig. 23 and 24. The mechanical focus mechanism may be controllable to adjust the position of the lens in at least one of the two camera modules (e.g., as generally shown in fig. 23). Additionally or alternatively, the mechanical focus mechanism may be controllable to adjust the position of the carrier (e.g., as generally shown in fig. 24).

Fig. 23 is a schematic diagram of elements of a test apparatus having an auto-focus function. As shown in fig. 23, the test apparatus can move the camera module up or down along the Z-axis (e.g., by a motorized rail, ultrasonic motor drive, or stepper motor). By adjusting the vertical position of the camera module, the testing device can adjust the distance between the camera module and the magnifying element.

FIG. 24 is a schematic diagram of the components of another test apparatus with an auto-focus function. As shown in fig. 24, the test strip tape assembly can be moved upward or downward along the Z-axis by the test device. By adjusting the vertical position of the test strip device, the test device can adjust the distance between the camera module and the amplifying element.

During the autofocus operation as shown in fig. 23 or fig. 24, the camera module and the supplemental lens remain as a single module. In other words, the distance between the camera module and the supplemental lens remains unchanged during the autofocus operation as shown in fig. 23 or fig. 24.

FIG. 25B is a schematic cross-sectional view B-B of the testing apparatus of FIG. 25A including a switch and a motor. The B-B cross section of test apparatus 1900 in FIG. 25B shows various elements of the test apparatus. The test device 1900 includes a switch 2550 to detect a collection vial 2510 inserted into the test device 1900. When the collection bottle 2510 is inserted, the switch 2550 is activated. The collection vial 2510 is then communicated to the testing device 1900 via switch 2550. Based on the time period that the switch 2550 is activated, the testing device may determine the time period that the collection bottle 2510 remains inserted.

The testing device 1900 further includes a motor 2560 for shaking, vibrating, or rotating the collection vials 2510 to mix the sample in the collection vials 2510. The testing device 1900 may include a camera 2570 to determine whether the sample has liquefied based on captured images of the sample in the collection vial 2510.

FIG. 26B is a cross-sectional view of the testing device including the flexible element shown in FIG. 26A. The B-B cross section of test apparatus 1900 in FIG. 26B shows various elements of the test apparatus. The testing device 1900 includes a moving element 2680 (e.g., a resilient element) at the bottom of the slot for movably receiving the collection vial 2610. For example, the moving element 2680 may include a spring that may spontaneously return to its normal shape after being contracted or deformed. As the collection bottle 2610 is inserted into the slot, the moving element 2680 is compressed. The light sensor 2690 (or other type of distance sensor) is responsible for detecting the distance between the light sensor 2690 and the bottom of the collection bottle 2610. Based on the distance between the light sensor 2690 and the bottom of the collection vial 2610, the testing device 1900 can determine the weight or volume of the sample contained in the collection vial 2610. For example, the distance between the light sensor 2690 and the bottom of the collection vial 2610 can be inversely proportional to the weight or volume of the sample contained in the collection vial 2610.

In some other embodiments, testing device 1900 may include a sensor on the top of collection bottle 2610. The sensor may be responsible for detecting the distance between the sensor and the top of the collection bottle 2610. The weight or volume of the sample contained in the collection vial 2610 can be determined based on the distance, as the volume or weight can be proportional to the distance between the sensor and the top of the collection vial 2610, for example. Then, based on the weight or volume of the sample, the testing device 1900 may determine a period of time to wait for the sample in the collection vial 2610 to liquefy. The testing device 1900 further includes a motor 2660 for shaking, vibrating, or rotating the collection vial 2610 in order to mix the sample in the collection vial 2610.

In some embodiments, the camera module of the testing device may include a light field camera (not shown) that captures the intensity and direction of the light. The light field camera may include a micro-lens array or a multi-camera array in front of the image sensor to detect directional data. Using the direction data of the light, the camera module can capture clear images at a wide range of focal planes. Therefore, the test device using the light field camera may not need an auto-focus function to finely adjust the distance between the camera module and the magnifying element.

In view of the above, the device of the present invention is suitable for testing male fertility and/or female fertility.

The present invention provides a method of testing male fertility using the device of the present application. The method comprises the following steps: applying a biological sample from a male subject to a first holding area and a second holding area of a carrier; inserting the vector into the device; determining the acidity of the sperm according to a first analytical treatment routine; determining one or more characteristics of sperm selected from the group consisting of: concentration of sperm, motility of sperm and morphology of sperm; and analyzing the results to determine male fertility.

The present invention also provides a method of testing female reproductive hormones using the device of the present application. The method comprises the following steps: applying a biological sample from a female subject to a first holding area of a carrier; inserting the vector into the device; and determining a concentration level of one or more types of female hormones, such as Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH), or Human Chorionic Gonadotropin (HCG).

The invention further provides a method for testing fertility in a pair of male and female subjects. The method comprises the following steps: applying a biological sample from a male subject to a first holding area and a second holding area of a first carrier; inserting a first vector into the device; determining the acidity of the sperm according to a first analytical treatment routine; determining one or more characteristics of sperm selected from the group consisting of: concentration of sperm, motility of sperm and morphology of sperm; applying a biological sample from a female subject to a holding area of a second carrier; inserting a second vector into the device; determining a concentration level of one or more types of estrogen; and analyzing the results of the male and female biological samples.

FIG. 27 is a flow chart of a processing routine for analyzing a semen sample of a male client or patient. A system for analyzing a semen sample may include a testing machine (e.g., testing device 1900), a mobile device, and a cloud server. FIG. 28 is a flow chart of a process routine for analyzing LH or HCG in a female client or patient. A system for analyzing LH or HCG may include a tester (e.g., test device 1900), a mobile device, and a cloud server. The flow charts of fig. 27 and 28 show steps performed by the tester, the mobile device, and the cloud server and data passed between the tester, the mobile device, and the cloud server.

In some embodiments, the method for testing sperm comprises the steps of: obtaining a device for testing a biological sample, applying a sperm sample to a sample holding area, recording video data or images of the sperm sample; determining a sperm count of the sperm sample based on the recorded video data or the at least one frame of recorded images; and determining sperm motility of the sperm sample based on the recorded video data or the recorded image.

In a related embodiment, the method further comprises: waiting a predetermined period of time for liquefaction of the sperm sample before applying the sperm sample to the sample holding area.

In another related embodiment, the method further comprises: placing a mobile device comprising a camera element on top of the device such that the camera element is aligned with a magnification element and a sample holding area; and receiving, by the mobile device, the optical signal from the sperm sample amplified by the amplifying element in the sample holding area.

In yet another related embodiment, the method further comprises: the sample holding area is illuminated by a lateral illumination device disposed on one side of the carrier of the device or a vertical illumination device disposed on top of or below the carrier of the device.

In yet another related embodiment, the method further comprises: directing the light beam from the lateral illumination device through a carrier made of a transparent or translucent material; and reflecting the light beam to the sample holding area by a plurality of light reflection patterns included in the carrier.

In yet another related embodiment, the method further comprises: the disposable testing device is inserted into a base that includes a camera element for recording video data of a sperm sample or a form-fitting frame for holding a mobile device that includes a camera element for recording video data of a sperm sample.

In yet another related embodiment, the method further comprises: extracting at least one frame from recorded video data of the biological sample; identifying a plurality of sperm from the at least one frame; and calculating the number of sperm based on the number of identified sperm and the area recorded by the at least one frame.

In yet another related embodiment, the method further comprises: analyzing the shape of the identified sperm; and determining a morphological level based on the identified shape of the sperm.

In yet another related embodiment, the method further comprises: extracting a series of video data frames from the recorded video data of the sperm sample; identifying a plurality of sperm from the series of frames of video data; identifying a movement track of the sperm based on the series of video data frames; determining the moving speed of the sperm based on the moving track of the sperm and the time period captured by the series of video data frames; and calculating sperm motility based on the speed of movement of the sperm.

In yet another related embodiment, the method further comprises: the video data or image of the sperm sample is further magnified by a magnifying lens.

In some embodiments, a method for testing sperm using a system for testing a biological sample, comprising: inserting the device into the base element; recording video data of the sperm sample in the sample holding area by a mobile device secured in a form-fitting frame of the base member; determining a sperm count of the sperm sample based on the at least one frame of recorded video data; and determining sperm motility of the sperm sample based on the recorded video data.

In a related embodiment, the method further comprises: the video data of the sperm sample is further magnified by a magnifying lens.

In some embodiments, a system for testing a biological sample includes a disposable device and a base element for testing a biological sample. The disposable device includes a sample carrier containing a sample holding area and a removable cover placed on top of the sample holding area. The base member includes an insertion port for inserting the disposable device into the base member, and a camera member for capturing an image of the sample holding area, the camera member including an image sensor and an optical lens module. In a related embodiment, the optical lens module may have a linear magnification ratio of at least 0.1.

Fig. 31 illustrates an additional example carrier 3105 having one or more visual markers 3117(1) -3117(5) (or may be collectively referred to as visual cues 3117) that may be used to control the analytical processing routines performed by the test apparatus (e.g., the test apparatus shown in fig. 21C or fig. 22). As shown in fig. 31, visual cue 3117 may be in (or near, in some additional or alternative embodiments) a carrier 3105 holding region (e.g., holding region 3115B).

As previously described (e.g., with respect to fig. 29), the processor may perform different analysis processing routines based on visual cues on a carrier. For example, in some particular embodiments, image recognition may be performed and the image of the holding area processed and different analysis processing routines may be performed based on visual cues from the results of the image recognition. In some embodiments, the visual cue of the carrier may be a shape of a particular holding area. Additional visual cue embodiments may include graphical patterns, visual indicia, one-dimensional barcodes, multi-dimensional pattern codes (e.g., QR codes), and the like.

In some specific examples, as shown in fig. 31, any visual indicia (e.g., visual indicia 3117(1)) may be a specific small graphic pattern that may be imprinted, attached, or otherwise marked on holding area 3115B in carrier 3105. In the example of fig. 31, the visual indicia 3117(1) (3117) (5) are all identical or substantially similar patterns, however in other examples (not shown for simplicity) they need not be identical and each visual indicia may have a unique shape, size, pattern, etc. In one or more implementations, the visual cue 3117 (e.g., visual indicia 3117(1) -3117(5)) is of a size that is not perceived by humans but can be recognized by a camera module (e.g., the second camera module 2230B of fig. 22 or the camera module 2130 of fig. 21C) after magnification by a microlens. In some implementations, the visual indicia 3117(1) - (3117) (5) are less than 15 micrometers (μm), and further, the visual indicia 3117(1) - (3117) (5) may be configured such that their locations collectively form a pattern (e.g., a predetermined configuration). In addition or as an alternative to the characteristics specific to the visual indicia (e.g., size, shape, color, and/or position), such a collective pattern formed by the positions of each visual cue may be a recognizable cue that is used to control the function of the test device (e.g., whether and subsequently what analysis processing routine is performed). Such an aggregate pattern may be based on the absolute position of the visual markers (e.g., at the holding area) and/or the relative position of the visual markers (e.g., from their respective neighboring markers). In fig. 31, the set pattern shown by the visual markers 3117(1) - (3117) (5) is that the visual markers 3117(1) - (3117 (4) are each disposed at one of the four corners (image captured by camera) and the visual markers 3117(5) are disposed at the center, each of the visual markers being uniformly distributed. Some additional or alternative embodiments provide that each (or each group of) particular visual cues (or indicia) of the visual cue may represent a different analysis function to be performed.

In view of the above, the test device herein may utilize visual cues (e.g., in or near the holding area) on the carrier to control the functionality of the test device and adaptively perform analytical processing routines based on the visual cues. In some embodiments, the visual cue may be used to confirm whether the carrier is an authorized carrier (e.g., properly authorized and manufactured under certain specifications and in accordance with applicable quality standards). In other examples, visual cues may be used to control the testing device to perform calculations in what mode (e.g., male or female, laboratory or home, high accuracy or short time, using a battery or plugged in). Further, providing visual cues in some embodiments may be used to control access to certain functionality of the test device, which provides the ability to flexibly order the services provided by the test device based on customer identity, geographic location, and the like.

Fig. 32 is a flow diagram of another example of a processing routine 3200 that may be implemented by the testing apparatus disclosed herein (e.g., in fig. 21C or 22) to perform an analysis processing routine based on visual cue adaptation. With continuing reference to FIG. 31, the process routine 3200 is described below. It should be noted that in the example of processing routine 3200, the visual cue is used to execute a carrier authentication application, although the processing routine may be equally applicable to executing other applications (e.g., as described with respect to fig. 30). For example, in some applications other than carrier verification, the processor may perform different sets of analysis processing routines based on different visual cues.

First, in step 3202, a carrier inserted through the opening is received in a receiving mechanism of the testing device, and a sensor (not shown for simplicity) may notify a processor, which will cause a camera module built into the testing device to capture an image of one or more carrier holding areas. At step 3204, the processor may identify visual cues in the carrier using the captured images, such as those based on known image analysis techniques or disclosed herein. As discussed above, the visual cue may include a number of visual indicia, each of which may or may not be the same size, shape, pattern, color, etc. (examples shown in FIG. 31). The visual indicia may together further present a pattern (e.g., from their location). The processor may then compare the visual cues (e.g., unique sizes, shapes, locations, or aggregate patterns) to predetermined visual cues (e.g., stored in local memory and/or in a cloud database (which may be operated or controlled by the manufacturer of the test device or other administrator).

In step 3206, the processor selectively performs a set of analysis processing routines on the captured image of the holding area based on the recognition result of the visual cue. If the recognition of the visual cue returns to positive (e.g., in response to the carrier holding area having the predetermined visual cue), then the processor proceeds with subsequent steps that may include selectively capturing additional images (or video data) for analysis (step 3208) and performing a corresponding set of analysis processing routines on the images (step 3210). On the other hand, if the recognition result returns negative (e.g., in response to a carrier holding area not having a predetermined visual cue), the processor causes an alternative action (e.g., displaying an error code) to reflect the non-recognition of the visual cue and does not perform any analysis processing routine on the image (step 3212). After the set of analysis processing routines is executed, the processor may continue to determine a result for the biological sample based on the results of the analysis processing routines as previously described.

In addition, it is noted that conventional computer-assisted sperm analysis (CASA) relies on the experience of large microscopes and operating technicians to determine sperm parameters. There is some computer software assistance to supplement the experience of the technician and to normalize the analysis results. However, due to the difference between the lens and the sensing module, the blurred image often seriously affects the effectiveness of the auxiliary software, resulting in inaccuracy of related functions (such as counting the number of sperm).

In addition, human semen examination and processing laboratory manuals are published by governing bodies such as the World Health Organization (WHO) WHO specify a minimum number of samples (e.g., 200 sperm) to be evaluated to determine sperm concentration, sperm motility, and sperm morphology. Existing computer-assisted sperm analysis image-based analyses generally lack automated sampling or require manual manipulation to acquire multiple fields of view to achieve world health organization specifications and reduce sampling errors in the analysis, and alternatively, if sampling is performed repeatedly with only a single field of view, the time spent in repeating the procedure tends to become too long to be achieved on a large scale in order to achieve satisfactorily low sampling errors.

Fig. 33 is a flow diagram of an example processing routine 3300 that can be implemented by the test device herein (e.g., fig. 21C or fig. 22) for better results (e.g., better analysis accuracy or efficiency). The processing routine 3300 may be an alternative or supplemental processing routine to the processing routine described herein (e.g., the processing routine shown in fig. 16).

First, step 3310, e.g., after insertion of a carrier cartridge carrying the biological sample or having been exposed to the biological sample (as introduced above), such introduced device may utilize a camera module to capture one or more images (or collectively, collective images (images)) of the carrier cartridge holding area. In some alternative embodiments (e.g., with respect to the form of fig. 29 or fig. 31), the device may identify (step 3320) the visual cue on the carrier from the captured aggregate image of the holding area. In these alternative embodiments, the device may perform a set of analysis processing routines on the captured aggregate image based on the recognition of the visual cue.

In step 3330, the apparatus divides the captured set of images into a plurality of blocks. In some embodiments, the tiles may be polygonal. More particularly, some implementations indicate that the tiles may be triangular, rectangular, square, pentagonal, hexagonal, and so forth. These (block) shapes may have at least one side of 0.05 mm. In one or more embodiments, the tiles are square and have dimensions of 0.05 millimeters by 0.05 millimeters. It is noted that, depending on the particular implementation, the number and size of the blocks may be adjusted according to the resolution of the camera module. An example of an image indicating one holding area is partitioned into blocks (e.g., block 3402) is illustrated in FIG. 34. It should be noted that, to facilitate discussion of the disclosed techniques herein, the captured aggregate image is considered "partitioned" into blocks; however, in one or more implementations, it should be appreciated that the processor need not actually perform mathematically partitioning operations to perform this technique during computer operation time (or during normal operation); rather, the resulting blocks or grids may be predetermined, logically pre-connected, code-preset, or pre-configured on the camera controller and/or processor of the device, such that the need for performing operations associated with dividing the aggregate image into blocks may be reduced, or in some instances, eliminated altogether.

In step 3340, the example device selects from a plurality of blocks, candidate blocks, for analysis. According to one or more embodiments, the candidate block is selected based on a number of factors, such as the degree of focus of a block and/or the normality of a block.

More specifically, in many implementations, the apparatus may determine (step 3342) the focus level of each of the multiple blocks, such that each block is capable of having a corresponding focus level measurement. This degree of focus may be determined based on one or more focus measurement functions. Depending on the implementation, the focus measurement function adopted may include one or more of: a variance type, a difference coefficient sum type, a laplacian energy image type, and/or a side gradient intensity maximization type.

After determining the focus level for each tile, in some embodiments, the device compares the focus level for a tile to a minimum focus level threshold. In one or more implementations, a tile may be selected as a candidate tile only if the degree of focus meets (e.g., reaches or exceeds) a minimum degree of focus threshold. In addition, the device may also be labeled or labeled with a block. This apparatus in one or more embodiments indicates that a block is marked or labeled (e.g., for further analysis or tracking identification) only if a minimum focus threshold is met. The marking or labeling may be done continuously or randomly. The selection of a portion of the candidate block is illustrated in fig. 35, where blocks are randomly labeled and blocks exceeding the minimum focus level threshold are initially selected as candidate block 3510.

Next, the device may execute an image processing program on the selected blocks to determine characteristics of the selected blocks (step 3344) to determine normality of a block, e.g., to see if the block is "" normal enough "" to allow further analysis. In some examples, the blocks selected for normality determination are those that have been preliminarily selected as candidate blocks (e.g., those that meet a minimum focus threshold, as described above). In some examples, the characteristic used to determine normality at this stage is cell number (e.g., sperm number). In a particular example, the apparatus may perform an image processing routine on blocks that satisfy a minimum degree of focus (i.e., they are sufficiently focused) to determine the number of cells (sperm) within a block for each sufficiently focused block. The image processing program may include binarization (and in some implementations, with adaptive thresholding) to identify parts of the block that are likely to be sperm as foreground and the rest of the block as background, and the device may determine the number of cells (sperm) after the image processing program. In one or more embodiments, the number of cells of the candidate block can be determined based on the ratio of areas with sperm to areas without sperm (e.g., extrapolated using a table associated with known cell count ratios).

Thereafter, the device can calculate statistics (e.g., mean and standard deviation) for all remaining candidate blocks (e.g., those that meet the minimum focus threshold). After the statistics are computed, the device is able to determine (step 3344) normality of the particular block by statistically comparing one or more characteristics (e.g., number of sperm) of the particular block with all remaining candidate blocks. In some embodiments, the selection of the candidate block is continued only if the normality of the particular block satisfies a normality condition. Taking the number of sperm as an example, in various embodiments, a block is considered "sufficiently normal" (e.g., satisfying a normality condition) when the number of sperm in the block is within a predetermined number of standard deviations of the average for a plurality of blocks. In one or more implementations, normality is required to be within two standard deviations (in terms of mean). In other implementations, normality requirements are one or three standard deviations, or other suitable statistical techniques, that reflect a comparison of normality of a block to other blocks. Fig. 36 illustrates the results after the image processing program (e.g., adaptive threshold binarization) and cell number determination. Note that in fig. 36, a flag indicating that the estimated cell number of each candidate block replaces it is shown.

In addition, the apparatus may determine (step 3346) whether the target number of cells to be analyzed has been reached. In particular, one or more embodiments of the disclosed apparatus are capable of maintaining a total cell number, and for each block selected as a candidate block, the apparatus adds the corresponding cell number for that block to the total cell number. The device may use the target number of analysis cells to control the number of biological samples to be analyzed, and the number may be configurable depending on the implementation. This quantity (number) can be customized according to the laboratory manual and the standard for testing a particular biological sample. In some embodiments, the target number of cells to be analyzed is two hundred (200). In some embodiments, the selection of candidate blocks is completed when the total cell mass reaches a target number of cells to be analyzed. That is, according to at least some embodiments disclosed herein, selection of candidate blocks can be performed (e.g., in a random manner) on blocks that meet the normality requirements for a degree of focus threshold and a total cell count that achieves a target number of cells to be analyzed.

After selecting the candidate block, the introduced device may determine one or more characteristics of the biological sample by analyzing the selected candidate block (e.g., by one or more of the techniques introduced herein), at step 3350. In at least some embodiments, the biological sample is semen, and one or more of the biological samples in the candidate block are determined to have a specific desire, which includes one or more of: cell number (or concentration, which can be inferred from cell number), motility, or morphology. In some examples, the device is further configured such that, after the set of analysis processing routines is executed, a determination can be made as to the outcome (e.g., fertility) of the biological sample based on the results of the analysis processing routines.

Furthermore, it is observed here that it is often difficult (especially in large quantities and when costs have to be controlled) to perfectly manufacture assemblies of ophthalmic lenses such as microscopic lens assemblies and/or magnifying lens assemblies mounted on the test devices introduced here. Imperfections in the lens, such as impurities, or imperfections in the lens itself (e.g., clarity, refraction, focus, and others), can adversely affect the accuracy of the test apparatus. Thus, described herein are techniques for correction and qualification to mitigate lens imperfections and to further improve the analytical accuracy of the test devices disclosed herein.

FIG. 37 is an example flow diagram 3700 of a calibration processing routine that may be implemented by a test device disclosed herein (e.g., FIG. 21C or FIG. 22) to achieve improved results. This processing routine 3700 may be an alternative or supplemental processing routine to the processes disclosed herein, such as the processing routine illustrated in fig. 16.

First, in step 3710 (e.g., after the carrier cassette is inserted), the introduced device can utilize a camera module to capture one or more images (or collectively, collective images (images)) of the carrier cassette holding area. In alternative embodiments (e.g., those described with respect to FIG. 29 or FIG. 31, the device may identify 3720 visual cues on the carrier from the collective image captured in the holding area. in such alternative embodiments, the device may perform a set of analysis processes on the captured collective image based on the results of the identification of the visual cues.

More specifically, in some implementations, the carrier box herein may act as a dedicated virtual box that may be used to trigger the calibration process routine. For example, a special virtual box may carry one or more special graphic patterns (e.g., as described below with respect to fig. 38), which, after the visual cue recognition processing routine (e.g., in step 3720), can trigger the testing device to enter the calibration mode. In another example, a dedicated virtual cartridge can carry a dedicated test sample (e.g., as described below with respect to fig. 41), and a user can manually cause (e.g., via a user interface on a panel or a remote test device) the test device to enter the calibration mode. In many instances, the virtual cartridge may include an electronic (e.g., a Radio Frequency Identification (RFID)) or a mechanical feature (e.g., a special shape or mechanical protrusion) that can trigger the calibration mode.

FIG. 38 shows a test carrier carrying a visual cue or an image pattern that can be used to calibrate or verify the test apparatus disclosed herein. In one or more embodiments, the visual cue comprises an image pattern that enables the testing device to recognize as a trigger to enter the calibration mode. The test device can then capture images of the image pattern using the camera module and perform self-diagnostics to self-calibrate the results from the captured images. The visual pattern should be easy to recognize (and not easily misrecognizable). As illustrated in fig. 38, the visual pattern in the example includes repeating (e.g., every 0.08 mm, i.e., repetition rate or "pitch"), larger (e.g., 0.02 mm by 0.02 mm), and generally regular shapes. The visual cue may further comprise one or more repeating linear patterns. In the example illustrated in fig. 38, this linear pattern includes a set (e.g., three) of horizontal lines and a set (e.g., three) of vertical lines. In some embodiments, the lines have a resolution of 200 line pairs per millimeter (LP/mm) or higher. In the specific example of FIG. 38, the lines have a resolution of 500 LP/mm. It should be noted that level and/or vertical lines are examples of visually linear patterns suitable for assisting the test apparatus to perform self-diagnostics of the optical characteristics and performance of the optical instrument (e.g., microlens) specifically mounted to the test apparatus itself; other suitable visual patterns may be substituted for the illustration in fig. 38. For example, in some embodiments, an "E" shaped pattern or equivalent may be used as the visual linear pattern instead of parallel lines. For example, in some implementations, solid and dashed lines can be used for the visual linear pattern.

Continuing with routine 3700, although the calibration mode has been initiated at step 3730, after the aggregate image of the carrier is captured (e.g., at step 3710), the device can segment the captured aggregate image into a plurality of blocks (similar to step 3330, discussed above). In some embodiments, the tiles may be polygonal. More particularly, some implementations point out that the blocks can be triangular, rectangular, square, pentagonal, hexagonal, and so forth. These (block) shapes may have at least one side of 0.05 mm. In one or more embodiments, the tiles are square and have dimensions of 0.05 millimeters by 0.05 millimeters. It is noted that, according to a specific implementation, the number and size of the blocks can be adjusted according to the resolution of the camera module. In one or more implementations, the above-mentioned pitch (e.g., the visual pattern regular self-repeat rate) can correspond to the number of patches that the aggregate image can be segmented. In some embodiments, the pitch may correspond to the number of blocks into which the aggregate image can be segmented by the test device.

At step 3740, the device in the example is able to execute the calibration/self-diagnostic routines, e.g., at each block. The calibration routine should generally be one or more steps that enable the testing device to autonomously self-diagnose the quality of the optical modules (e.g., including microlenses, camera modules) currently mounted on the testing device itself. In one or more embodiments, the test device may determine (at 3742) the degree of focus for each block, for example, by using one or more focus measurement functions. Examples of the focus measurement function may include a variance type, a difference coefficient sum type, a laplacian energy image type, and/or a side gradient intensity maximization type. Next, at step 3744, the test instrument determines whether a block satisfies a focus level, such as the minimum focus threshold discussed above. Additionally or alternatively, the test device can compare the retrieved results to one or more expected results (at step 3746). For example, the processor of the test instrument accesses one or more images pre-installed in memory (e.g., not captured using a camera, e.g., passed or encoded with a default installation), compares the captured images, and determines whether the suspected captured image quality in the block meets a minimum criterion. The pre-installed image or images should be representative of the visual pattern applied for correction. At step 3746, the testing device can compare and inspect example image quality parameters, including color distortion, pattern distortion, sharpness defects, and/or other image defects.

FIG. 39 is an exemplary image of the visual enhancement 38 captured by the testing apparatus disclosed herein, with the image quality generally being better at the lower left and worse at the upper right. Fig. 40A and 40B are specific example illustrations of two different image qualities of the captured image in different blocks in fig. 39. In some embodiments, for example, where the pitch is consistent with the number of blocks that can be used to segment the aggregate image, fig. 40A and 40B can each represent a block. As the figure illustrates, the image quality in the block of fig. 40A is better than that of fig. 40B because the image is sharper and more focused.

Referring back to the processing routine 3700, in step 3750, the result of step 3740 (e.g., whether the block meets the minimum image quality requirement, such as the minimum focus level) is recorded in a computer readable storage medium (e.g., non-transitory, such as a flash memory) incorporated in the testing apparatus (not shown for simplicity). The knowledge obtained from the calibration process routine may, for example, be utilized when the test device is subsequently in normal operation. In one or more embodiments, during normal operation (e.g., in step 3340, as discussed above), the testing device may automatically skip or ignore blocks that fail to meet minimum image quality requirements during calibration or self-diagnosis. In this way, the test device disclosed herein can mitigate adverse effects of lens imperfections and increase analytical accuracy.

FIG. 41 illustrates an exemplary image of a test carrier carrying a test sample that may be used to calibrate or verify the test apparatus disclosed herein. This technique can be applied to one or more of the foregoing embodiments where a dedicated virtual cartridge can carry a dedicated test sample and the calibration mode can be activated by triggers other than visual patterns (e.g., manually by a user, or by mechanical features or Radio Frequency Identification (RFID) on the virtual cartridge). Certain embodiments indicate that the test sample should be in the form of an aqueous medium (e.g., a liquid solution) containing small test particles, such as test particles 4102 illustrated in fig. 41. The microparticles may be made of any suitable material, including, for example, polymers. A particular example material of particles 4102 is latex. The size of the particles may be adapted to the specific application. In certain implementations, the size of the particles can be similar to the size of those cells, such as sperm. Example sizes of particles can range from 0.5 microns to 50 microns in diameter. In the examples, the particles are 5 microns in diameter. When the test particles are used as a sample, the test apparatus can perform calibration/self-diagnosis as in the 3700 process routine without the step 3720, and self-diagnose the quality of the optical module currently installed in itself. In some of these implementations, the test device may pre-mount images of the test particles (e.g., not captured by the camera, e.g., by being transmitted or otherwise encoded) in memory, such as discussed above, for comparison and calibration purposes.

Fig. 42A and 42B illustrate different image qualities in different blocks of the captured image of fig. 41. As illustrated, the image quality in the block of FIG. 42A is better than that of FIG. 42B because the image is sharper and more focused. Similar to the discussion above, with respect to step 3750, knowledge of the baseline image quality for each block may be used, for example, when the test apparatus is subsequently operating normally. For example, some embodiments of the test equipment may automatically skip or ignore blocks that fail to achieve the minimum image quality condition during calibration or self-diagnose. In this way, the test apparatus disclosed herein can mitigate adverse effects of lens imperfections and improve analysis accuracy.

In some embodiments, a visual cue (e.g., as discussed in relation to FIG. 31) may indicate that the test equipment determines the status of the carrier or sample holding area (e.g., the area is dry or wet, or carries a good sample). FIG. 43 is a flow diagram of an example of an implementable processing routine 4300 for verifying that the sample-holding region carries a biological sample by the test apparatus disclosed herein (e.g., in FIG. 21B or FIG. 22).

Step 4310 (e.g., after a carrier cartridge is inserted), the device may acquire one or more images (or merge referred to as aggregate images) of the holding area of the carrier cartridge using the camera module. Step 4320 the device may identify a visual cue on the carrier based on the captured collective image of the holding area. Visual identification includes step 4342 verifying whether the holding area carries a biological sample based on the manner in which the visual cue is displayed in the captured aggregate image. The identifying further includes selectively causing the processor to perform the set of analysis processing routines on the captured aggregate image based on the verification result at step 4346. The device can perform the set of analysis processing routines on the captured aggregate image according to the recognition result of the visual cue (step 4330).

Fig. 44A-44B show examples of captured aggregate images of an empty or dry sample-holding area. Fig. 45A-45B illustrate an example of a sample holding area loaded with an captured image of a fluid carrying a biological sample. In these particular examples, each captured image shows a visual cue of a micro-graphic pattern. The shading of the visual cues is clearly visible in the empty or dry sample holding area. The fluid carrying the biological sample will cause the sample holding area to show a different refractive index (reactive index) compared to an empty carrier cartridge, and will therefore influence the extracted shape of the visual cue. The shadows of the visual cues will be faded by refraction. For example, an extracted shape with a visual cue of a biological sample will form an optical distortion (optical distortion) compared to a template (the extracted shape without any visual cue of the sample as shown in fig. 44A-44B). Thus, by comparing the captured shape of the visual mark with the template and evaluating the distortion, such as determining the boundary thickness of the mark or the shadow region, the processor can determine whether the holding area is actually loaded with the biological sample to be tested. Here, the distortion threshold represents the degree of optical distortion that shows the presence of the biological sample in the holding area. In particular, the degree of distortion may be compared to a predetermined threshold to verify whether the carrier carries a biological sample. In many implementations, different thresholds may be defined based on the algorithms employed and/or heuristic algorithms (heuristics), e.g., algorithms such as pixel gradient direction and intensity detection may be used to determine the corresponding threshold.

In some embodiments, the template may be generated by capturing images of empty carriers during a deployment phase and storing image data representing visual cues of the captured images prior to detecting any biological sample. In some embodiments, the template may be stored in the device to reduce the amount of operands that need to be executed during the configuration phase. In some embodiments, the visual cue may be compared to the stored images to determine the dissimilarity of the two. When the difference is below a predetermined threshold, the processor may determine that the holding area carries the biological sample.

If the processor verifies that the sample-holding region carries a valid sample, the processor performs an analysis of the captured image. However, if the processor cannot verify based on the comparison that the sample-holding region carries any valid biological sample (e.g., when a fluid sample is required for the analysis task to be performed (recognizing the shape of the holding region as described above in some examples), rather than a fluid), the processor does not perform any image analysis. Instead, in some embodiments, the device notifies the user that an invalid sample is being provided. The notification may be shown with a signal or a flag indicating that no biological sample has been provided.

In some cases, as previously described, CASA-based image analysis typically lacks automatic sampling or requires manual post-processing (post-processing) to obtain multiple fields of view to meet WHO specifications. To address this problem, the test apparatus may include a positioning mechanism operable to adjust the relative position of the carrier to the camera with or without human intervention so that images can be acquired with a plurality of adjacent fields of view.

FIG. 48 is a flow diagram of an example 4800 processing routine that may be implemented by the test equipment disclosed herein to improve results using multiple fields of view. At step 4810, the apparatus may utilize a camera module to capture a first image of the sample holding area of the carrier, for example, after the carrier cartridge is inserted. The step 4820 device identifies edges of the first image and the positioning mechanism of the step 4830 device adjusts the relative position of the carrier to the camera (by adjusting the carrier, the camera, or both) so that when the camera acquires the second image, the edges of the second image are adjacent to or aligned with the identified edges of the first image. The step 4840 device performs a set of analysis processing routines on the combined first and second images to determine one or more characteristics (e.g., cell number) of the biological sample. In some embodiments, when the edges of the second image are aligned with the edges of the first image, the two images may be used to compose a larger image. In some embodiments, the first and second images are from different perspectives only. The second image may be located adjacent to the first image or randomly at a position such that the second image does not overlap the first image.

In some embodiments, the positioning mechanism may determine multiple fixed positions of the camera. In some embodiments, the positioning mechanism may determine multiple positions of the carrier. For example, a multi-axis mobile platform (multi-axis mobile platform) as shown in fig. 49 may be used to adjust the carrier position and/or the camera position.

In some embodiments, a plurality of markers (e.g., visual cues as shown in fig. 31) may be placed on the test equipment (e.g., on the movable platform) so that they may be detected by the camera. After the camera determines the position of the mark, the camera starts scanning using the mark as a starting point. The remaining marks serve as a guide for the camera scan path, thereby enabling the testing equipment to adjust the camera to a fixed position automatically to capture images from multiple viewing angles. The scan path along with the mark may be random as long as no area is repeatedly scanned. The scan paths may also be in a certain order (e.g., clockwise or counterclockwise). For example, four markers may be used to mark four corresponding regions. The camera can capture each region along the mark sequentially or randomly without repetition. In some embodiments, the complex axis 4901 of the platform enables manual or automatic adjustment of the carrier and/or camera in the direction X, Y and the carrier platform.

In some embodiments, the positioning mechanism adjusts the relative position of the carrier with respect to the camera in such a way that when viewed from the camera, the plurality of images captured by the camera are sequentially clockwise or counterclockwise. For example, fig. 50A shows an example manner in which images are captured in a clock-sequential order viewed from a camera. In some embodiments, the positioning mechanism adjusts the relative position of the carrier to the camera in such a way that a plurality of images are acquired by the camera in sequential scans as viewed from the camera. Fig. 50B shows an example manner of acquiring images in sequential scanning.

In some embodiments, the first and second images are combined before performing further image analysis. In some cases, the first and second images may have overlapping portions due to adjustment of the camera or the carrier. The overlapping portions may be removed using image processing techniques to form a combined image.

As in fig. 50A to 50B, the edge of the first image may be a bottom edge of the first image and the edge of the second image may be a top edge of the second image. In some cases, the edge of the first image is a top edge of the first image and the edge of the second image is a bottom edge of the second image. The edge of the first image may also be the left side of the first image and the edge of the second image may be the right side of the second image. Similarly, the edge of the first image may also be the right side of the first image and the edge of the second image may be the left side of the second image.

As described with respect to fig. 16, the testing apparatus calculates the concentration of sperm based on the volume of the sample captured multiplied by the distance between the sample holding area and the bottom of the housing, however, due to manufacturing inaccuracies, the volume of the sample may not be as great because the distance between the sample holding area and the bottom of the housing may vary. To compensate for the aforementioned inaccuracies and determine the actual volume of the sample, the testing apparatus may include a camera module having a focus motor, such as a Voice Coil Motor (VCM), a ceramic piezoelectric actuator (ceramic piezoelectric actuator), a servo motor (servo motor) mechanism for a monocular camera focus mechanism or for a microscope, etc. The focus motor is operable to drive a camera lens to adjust a camera focus. Fig. 51A to 51B illustrate an example of an adjustable lens of a camera module. In FIG. 51A, the lens is at an initial or preset position, corresponding to the first focal length (focal length) of the lens. In fig. 51B, the lens is extended to its maximum length, corresponding to the second focal length of the lens. Two different positions of the lens form a distance L which can be divided into a plurality of steps (step), each step corresponding to a change in focal length. Therefore, by knowing the positions of the two target object lenses, the distance between the two target objects can be derived.

Fig. 52 shows an example of a configuration for determining the actual volume of the sample contained in the carrier. The housing 5210 is placed on top of the carrier 5220 prior to inserting the carrier 5220 to capture an image of the sample. A first visual cue 5217a (e.g., as shown in fig. 31) can be placed at the bottom of the housing 5210. The second visual cue 5217b can then be placed on a surface of the carrier 5220 that is in contact with the sample of the carrier 5220 or has been exposed to the sample of the carrier 5520.

Before analyzing the biological sample in the carrier, the apparatus determines a first focus position of the camera module 5230 with respect to the housing 5220. This step is performed by adjusting the lens to focus on the first visual cue 5217 a. The device can also determine the second focus position of the carrier 5210 by adjusting the lens to focus on the second visual cue 5217 b. The device can then determine the focal distance from the two focal positions and calculate the distance between the first surface of the housing 5210 and the second surface of the carrier 5220. Such a processing routine may be performed at a configuration stage prior to analyzing a batch of samples (e.g., the carrier and cover in the same batch should be fabricated using the same equipment to have the same characteristics). For more accurate sample reading, this processing routine may also be performed at the point of use if each carrier and cover have different characteristics.

In one specific embodiment, the lens of the camera module can support a focal length in a range of 50 μm to 550 μm. This range corresponds to the distance L between the two lens positions, as shown in fig. 51A-51B. In some implementations, the distance L may be divided into 1024 steps (step 0, step 1, step …, step 1023), each corresponding to a particular focal length (i.e., each step corresponds to (550-50)/1024 ═ 0.49 μm with a different focal length). In the configuration phase, the test equipment determines the focus position of the first visual cue 5217a in advance as step number 235. The device then determines the location of focus of the second visual cue 5217b as step number 295. The difference between the two steps (295-. From this distance and the area of the extracted sample holding area, the actual volume of the sample can be determined.

In some examples, the housing 5210 and the carrier 5220 can have a plurality of visual cues, located on the top or bottom surfaces. To obtain a more accurate reading, the measurements may be performed at different locations of the holding area. It is noted that the focal length and the ratio of the distances may vary from lens design to lens design. The ratio used in the specific example above is 1:1 (i.e. the focal length of the lens is equal to the distance between the housing and the carrier). Various ratios between the focal length and the distance may be supported, such as 1:1.2,1:1.5,1:2, etc.

Although some of the specific examples disclosed herein apply the disclosed techniques to sperm testing, one of ordinary skill in the art will readily appreciate that the disclosed techniques may be applied to testing various types of biological samples, such as semen, urine, synovial joint fluid, superficial tissues or cells, tumor cells, water samples, and the like. In addition, the techniques disclosed herein can also be applied to various analytical processing protocols, such as Sperm Chromatin Dispersion (SCD) or terminal deoxyribonucleotide transferase deoxyuridine triphosphate nicking end labeling (TUNEL) for detection of DNA fragments. More particularly, SCD may be performed in one or more implementations, and the processor of the test apparatus described herein may be configured to determine normality (normalcy) of sperm DNA fragments using the captured images. In some instances, the sperm head exhibits a large or medium halo (halo) under the SCD test (dimensions are known in the art), and the processor can determine that there is no problem with DNA fragments in the sperm sample being tested. Conversely, if the sperm head exhibits a small, no halo, or a degraded halo, the processor determines that the sperm sample being tested has a problem with DNA fragments. Further, TUNEL may be performed in one or more implementations. In one or more implementations, the processor of the test apparatus described herein may be configured to detect apoptotic DNA fragments using the captured collective image. The processor may be used to identify one or more TUNEL stained cells, to quantify apoptotic cells and/or to detect excessive DNA fragmentation in individual cells.

In one example aspect, an apparatus for testing a biological specimen includes a receiving mechanism for receiving a carrier. The carrier includes a holding area configured to carry a biological sample. The apparatus includes a camera module configured to capture an aggregate image of the holding area. The apparatus also includes a processor configured to identify visual cues on the carrier from the captured aggregate image of the holding area using the camera module, and perform a set of analysis processing routines on the captured aggregate image based on the identification of the visual cues. The identification of the visual cue comprises: verifying whether the holding area contains the biological sample according to the display mode of the visual cue in the captured set image, and selectively enabling the processor to execute a set of analysis processing routines on the captured set image according to the verification result.

In some embodiments, the processor performs the set of analysis processing routines on the captured collective image only after confirming that the holding area carries the biological sample. In some embodiments, the processor does not perform the set of analysis processing routines on the captured collective image if it is not confirmed whether the holding area carries the biological sample. In some embodiments, the processor also displays a predetermined signal or indicia associated with the non-verifiable retention area containing the biological sample.

In some embodiments, the visual cue is displayed in a manner that includes optical distortion of the visual cue. In some embodiments, the optical distortion produced by the visual cue corresponds to a difference in refractive index between the biological sample and air. In some embodiments, verifying whether the holding area carries the biological sample includes comparing an optical distortion of the visual cue to a distortion threshold, wherein the distortion threshold represents a degree of optical distortion that may represent the presence of the biological sample in the holding area. In some embodiments, verifying that the holding area carries the biological sample comprises comparing a difference between the visual cue and a stored image of the set of visual cues, and if the difference is below a predetermined threshold, then the holding area is certified as carrying the biological sample.

In some embodiments, the processor is configured to capture a stored set of visual cue images when the carrier is free of any biological sample during the configuration phase. In some embodiments, the visual cue is located in or adjacent to a holding area on the carrier. In some embodiments, the visual cue is identified when the visual cue exists in a predetermined shape or at a predetermined location. In some embodiments, the visual cue comprises a preset configuration of a plurality of visual cues.

In some embodiments, the device includes a housing. The components of the device are all enclosed within the housing. The external dimension of the shell is less than 27000 cubic centimeters. In some embodiments, the device further comprises a display enclosed within the housing. The processor is configured to display the determined final result on the display after the final result is obtained. In some embodiments, the display is configured to display a notification based on a result of the verification. In some embodiments, the notification includes a signal or flag indicating that a biological sample has not been provided. In some embodiments, the processor is further configured to determine a biochemical property of the biological sample using the set of analytical processing routines.

In another example aspect, an apparatus for testing a biological sample includes a receiving mechanism to receive a carrier configured to carry the biological sample or to have exposed the biological sample. The apparatus includes a camera module configured to capture a plurality of images of a holding area. The apparatus also includes a processor configured to adaptively select an analysis algorithm adapted to the movement characteristics of the biological sample under test using the camera module based on the plurality of images of the holding area, and perform a set of analysis processing routines corresponding to the selected analysis algorithm on the captured plurality of images to obtain a related analysis result of the biological sample.

In some embodiments, the movement characteristics are maintained substantially static or dynamic. In some embodiments, the processor is to adaptively select the analysis algorithm based on determining an amount of change between the first image and the second image in the plurality of images, and selecting the static algorithm or the dynamic algorithm based on the determined amount of change being less than or greater than a threshold. In some embodiments, the amount of change is determined from a rate of change of movement of the detectable target in the biological sample.

In some embodiments, the processor is to adaptively select an analysis algorithm based on the steps comprising: and judging the variation between the first image and the second image in the plurality of images, comparing the judged variation with a threshold, and selecting an analysis algorithm suitable for the movement characteristic according to the comparison result of the judged variation and the threshold. In some embodiments, the analysis algorithm is a static algorithm or a dynamic algorithm. In some embodiments, the threshold is a characteristic of whether the biological sample is substantially static or dynamic. In some embodiments, the processor is configured to determine mobility of the biological sample using an analysis algorithm. In some embodiments, the selected algorithm is a static algorithm when the determined amount of change is less than a threshold. In some embodiments, when the determined amount of change is not less than the threshold, the selected algorithm is a dynamic algorithm.

In some embodiments, the processor is configured to determine the trajectory of the biological sample using a dynamic algorithm. In some embodiments, the first and second images are acquired at least a predetermined time apart in time. In some embodiments, the predetermined time is in a range between 0.04 seconds and 10 seconds. In some embodiments, for a given number of image acquisitions in sequence, both the first image and the second image are acquired separately. In some embodiments, the sequentially given number of image acquisitions ranges from 2 to 600 images. In some embodiments, the apparatus further comprises a housing, wherein the receiving mechanism, the camera module, and the processor are all enclosed within the housing, and the housing has an outer dimension of less than 27000 cubic centimeters.

In another example, an apparatus for testing a biological sample includes: a receiving mechanism to receive a carrier, the carrier comprising a holding area configured to carry or have been exposed to the biological sample. The apparatus includes a camera module configured to capture a plurality of images of a holding area, including a first image and a second image, and a positioning mechanism operable to adjust a relative position of a carrier and a camera. The apparatus also includes a processor configured to utilize the camera module to identify an edge of the first image, cause the positioning mechanism to adjust the relative positions of the carrier and the camera such that when the camera acquires the second image, an edge of the second image is adjacent to or on the identified edge of the first image and a set of analytical processing routines is performed on the combined images of the first and second images to determine one or more characteristics of the biological sample.

In some embodiments, the positioning mechanism adjusts the relative position of the carrier and the camera in such a way that the plurality of images are sequentially acquired by the camera in a clockwise or counterclockwise direction when viewed from the camera. In some embodiments, the positioning mechanism determines that the relative position of the carrier and the camera is based on a plurality of markers detected by the camera. In some embodiments, the positioning mechanism adjusts the relative position of the carrier and the camera by: the camera acquires a plurality of images in the order of sequential scanning when viewed from the camera. The processor may be further configured to combine the first and second images to form a combined image. In some embodiments, the analysis processing routine for the combined image does not include overlapping portions of the first and second images. In some embodiments, the one or more characteristics of the biological sample include cell number.

In some embodiments, the edge of the first image is the bottom edge of the first image and the edge of the second image is the top edge of the second image. In some embodiments, the edge of the first image is at the top edge of the first image and the edge of the second image is at the bottom edge of the second image. In some embodiments, the edge of the first image is on the left side of the first image and the edge of the second image is on the right side of the second image. In some embodiments, the edge of the first image is on the right side of the first image and the edge of the second image is on the left side of the second image.

In some embodiments, to adjust the relative position of the carrier and the camera, the positioning mechanism adjusts the carrier to a next position for the camera to acquire subsequent images. In some embodiments, to adjust the relative position of the carrier and the camera, the positioning mechanism adjusts the camera to a next position for the camera to acquire subsequent images. In some embodiments, the processor automatically adjusts the relative position of the carrier and the camera after the camera acquires the first image.

In some embodiments, the positioning mechanism is configured to automatically adjust the camera to a plurality of fixed positions. In some embodiments, the positioning mechanism includes a multi-axis motion stage configured to adjust the carrier or camera to a plurality of positions. In some embodiments, the multi-axis motion platform is configured to enable manual adjustment of carrier position. In some embodiments, the multi-axis motion platform is configured to enable automatic adjustment of the carrier position. In some embodiments, the apparatus further comprises a housing. The receiving mechanism, the camera module and the processor are all packaged in the shell, and the appearance size of the shell is 27000 cubic centimeters.

In another example, a device for testing a biological sample includes: a receiving mechanism to receive a carrier, wherein the carrier comprises a holding area configured to carry the biological sample or to have been exposed to the biological sample. The apparatus includes a camera module configured to capture a collective image of the holding area. The camera module includes an operable focus motor for adjusting the focus of the camera. The apparatus includes a processor configured to determine a volumetric characteristic of a holding area based on operation of a focus motor using a camera module and to perform an analysis processing routine on at least a portion of the captured holding area image to determine one or more characteristics of the biological sample. One or more characteristics of the biological sample are adjusted by a processor according to the determined volumetric property of the holding area.

In some embodiments, the processor is configured to determine the volumetric characteristic of the holding region by operating the focus motor to focus on different locations of the holding region. In some embodiments, the processor is configured to estimate the depth of the holding region simultaneously when determining the volumetric characteristic of the holding region. In some embodiments, the processor is configured to perform a calculation to convert the estimated depth of the holding area into a volumetric characteristic of the holding area.

In some embodiments, the processor is configured to determine the volumetric characteristic of the holding region by focusing the camera on one of a housing or holding region to form a first focal point, measuring a first depth of the first focal point, focusing the camera on the other of the housing or holding region to form a second focal point, and measuring a second depth of the second focal point. In some embodiments, each of the housing and the holding area includes a visual cue for focusing of the camera. In some embodiments, the bottom surface of the housing is also marked with visual indicia. In some embodiments, the visual cue is disposed on a surface of the holding area that is in contact with or has been exposed to the biological sample. The visual cues may be made in microscopic in size.

In some embodiments, the first and second depths are measured in accordance with operation of a focus motor. The first depth and the second depth may be measured based on the number of steps (how many steps) in which the focus motor operates when reaching the first focus and the second focus from a preset starting point. In some embodiments, a total travel (travel) available for focus adjustment is preset, and the total number of available steps corresponds to the total travel available for focus adjustment. In some embodiments, the focus motor element includes a voice coil motor, a ceramic piezoelectric motor, and a monocular camera focus mechanism or microscope servo motor mechanism. In some embodiments, the one or more characteristics of the biological sample also include a concentration of the biological sample. In some embodiments, the biological sample is semen.

In some embodiments, during the configuration phase, a volumetric characteristic of the holding region is determined. In some embodiments, an analytical processing routine is performed on the biological sample during normal use. In some embodiments, the device further comprises a housing, the receiving mechanism, the camera module, and the processor all enclosed within the housing, the housing having an outer dimension of less than 27000 cubic centimeters.

It will be apparent to those skilled in the art that various modifications and variations can be made in the structure of the present invention without departing from the scope or spirit of the invention. In view of the foregoing, it is intended that the present invention cover modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.

Claims

1. An apparatus for testing a biological sample, the apparatus comprising:

a receiving mechanism to receive a carrier, wherein the carrier comprises a holding area, wherein the holding area is configured to carry the biological sample or has been exposed to the biological sample;

a camera module configured to capture a plurality of images of the holding area, the plurality of images including a first image and a second image;

a positioning mechanism operable to adjust the relative position of the carrier to the camera module; and

a processor configured to utilize the camera module to:

identifying edges of the first image;

causing the positioning mechanism to adjust the relative position of the carrier to the camera module such that the edge of the second image is aligned with the identified edge of the first image when the camera module acquires the second image; and

a set of analytical processing routines is performed on an image including the first image and the second image to determine one or more characteristics of a biological sample.

2. The apparatus of claim 1, wherein the positioning mechanism is caused to adjust the relative position of the carrier to the camera module such that the plurality of images are acquired by the camera module in a predetermined sequence when viewed from the camera module.

3. The apparatus of claim 1, wherein the positioning mechanism is caused to adjust the relative position of the carrier to the camera module such that the plurality of images are sequentially acquired by the camera module in a clockwise or counterclockwise direction when viewed from the camera module.

4. The apparatus of claim 1, wherein the positioning mechanism is caused to adjust the relative position of the carrier to the camera module such that the plurality of images are acquired by the camera module in a sequential scanning order when viewed from the camera module.

5. The apparatus of claim 1, wherein the positioning mechanism is caused to adjust the relative position of the carrier to the camera module such that the plurality of images are acquired by the camera module when viewed from the camera module.

6. The apparatus of claim 1, wherein the processor is further configured to combine the first image and the second image to form the image.

7. The apparatus of claim 1, wherein the analysis processing routine for the image does not include an overlapping portion of the first image and the second image.

8. The device of claim 1, wherein the one or more characteristics of the biological sample comprise cell number.

9. The apparatus of claim 1, wherein the edge of the first image is a bottom edge of the first image and the edge of the second image is a top edge of the second image.

10. The apparatus of claim 1, wherein the edge of the first image is at a top edge of the first image and the edge of the second image is at a bottom edge of the second image.

11. The apparatus of claim 1, wherein the edge of the first image is on the left side of the first image and the edge of the second image is on the right side of the second image.

12. The apparatus of claim 1, wherein the edge of the first image is on the right side of the first image and the edge of the second image is on the left side of the second image.

13. The apparatus of claim 1, wherein to adjust the relative position of the carrier and the camera module, the positioning mechanism adjusts the carrier to a next position for the camera module to capture subsequent images.

14. The apparatus of claim 1, wherein to adjust the relative position of the carrier and the camera module, the positioning mechanism adjusts the camera module to a next position for the camera module to capture subsequent images.

15. The apparatus of claim 1, wherein the processor automatically adjusts the relative position of the carrier and the camera module after the camera module acquires the first image.

16. The apparatus of claim 1, further comprising:

a housing, wherein the receiving mechanism, the camera module, and the processor are all enclosed within the housing, wherein the housing has an outer dimension of less than 27000 cubic centimeters.

17. The apparatus of claim 1, wherein the one or more characteristics of the biological sample comprise a concentration of the biological sample.

18. The device of claim 1, wherein the biological sample is semen.

19. A controller for controlling a device for testing a biological specimen, the device comprising:

a camera module configured to capture a plurality of images of the holding area, the plurality of images including a first image and a second image; and

a positioning mechanism operable to adjust the relative position of the carrier to the camera module; and is

Wherein the controller is configured to utilize the camera module to:

identifying edges of the first image;

20. The controller of claim 19, further comprising:

a housing, wherein the receiving mechanism, the camera module, and the controller are all enclosed within the housing, wherein the housing has an outer dimension of less than 27000 cubic centimeters.