KR102598764B1 - Microparticle counting apparatus and method for correcting brightness of image using calibration table - Google Patents

Abstract

The present disclosure seeks to provide an apparatus and method for counting fine particles. In one embodiment, the fine particle counting device determines the index exposure time, and when a value corresponding to the index exposure time exists in the calibration table, the fine particle counting device uses the calibration table to automatically focus on the index exposure time. Determine the exposure time and chamber exposure time, identify the insertion of the slide containing the alignment section and the chamber, and perform an automatic focusing procedure of the alignment section of the slide based on the autofocus exposure time determined using the calibration table. And, based on the chamber exposure time determined using the calibration table, an image of the chamber of the slide can be acquired.

Description

Micro particle counting device and method for correcting the brightness of an image using a calibration table {MICROPARTICLE COUNTING APPARATUS AND METHOD FOR CORRECTING BRIGHTNESS OF IMAGE USING CALIBRATION TABLE}

The present disclosure relates to a fine particle counting device and method for correcting the brightness of an image of a slide. More specifically, it relates to a fine particle counting device and method for correcting the brightness of an image of a slide chamber using a calibration table.

Cell counting refers to quantifying the number of cells and can be used in a variety of fields such as biology and medicine. For example, the number of blood cells obtained as a result of counting blood cells can inform the patient's health status, and in cell therapy, the cell count can be used to adjust the amount of cells to be administered to the patient. In addition, the concentration of pathogens such as bacteria and viruses in blood or body fluids can provide information about the progress of infectious diseases and the immune system.

Since cells are small in size and are cultured in large numbers even in a small amount of culture medium, cell counting is usually performed by extracting and counting a partial area, calculating the ratio of the extracted area to the remaining culture area, and then multiplying the number. Accordingly, a hemocytometer is used in which cells distributed on a counting grid among the cells distributed in the chamber of the slide are the cells to be counted. However, this manual cell counting method requires a lot of time and effort, and has the problem of producing very subjective results because cell separation may be different depending on the measuring researcher.

Recently, computer-based automatic cell counting methods have been used beyond these manual cell counting methods. The automatic cell counting method also uses a slide with a counting grid in the chamber, and the automatic cell counting equipment aligns the slides based on the grid and performs image-based automatic cell counting using a counting algorithm.

This automatic cell counting device must always perform counting using images with the same intensity to obtain reliable results. Factors that determine the intensity of an image acquired through a camera include brightness, contrast, gain, and exposure time. Among these, the easiest factor to control intensity without distortion is exposure time. The present disclosure seeks to provide a method for quickly adjusting exposure time for automatic brightness adjustment.

One embodiment of the present disclosure is to provide a fine particle counting device and method for correcting the brightness of an image of a slide chamber using a calibration table.

One embodiment of the present disclosure is to provide a fine particle counting device and method for correcting factors (eg, exposure time) for adjusting the brightness of an image of a chamber of a slide using a calibration table.

One embodiment of the present disclosure includes a memory storing one or more instructions; and at least one processor executing the one or more instructions stored in the memory. In one embodiment, the at least one processor determines the index exposure time by executing the one or more instructions, and if a value corresponding to the index exposure time exists in the calibration table, the at least one processor determines the index exposure time by using the calibration table. Determine an autofocus exposure time and a chamber exposure time corresponding to the index exposure time, identify insertion of a slide including an alignment portion and a chamber, and based on the autofocus exposure time determined using the calibration table, An automatic focusing procedure of the alignment unit of the slide may be performed, and an image of the chamber of the slide may be acquired based on the chamber exposure time determined using the calibration table.

In one embodiment, the at least one processor may identify a target image intensity value and determine an exposure time corresponding to the target image intensity value as an index exposure time before the slide is inserted.

In one embodiment, the at least one processor may determine the index exposure time based on a binary search algorithm.

In one embodiment, the index exposure time, the autofocus exposure time, and the chamber exposure time may be mapped to each other and stored in the calibration table.

In one embodiment, the at least one processor identifies that a value corresponding to the index exposure time is not in the calibration table, and selects the target based on an algorithm regarding the relationship between the autofocus exposure time and image intensity. determine a first autofocus exposure time corresponding to the image intensity value, and determine a first chamber exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the chamber exposure time and image intensity, The calibration table can be updated by associating the index exposure time, the first autofocus exposure time, and the first chamber exposure time with the calibration table.

In one embodiment, when at least one of the autofocus exposure time and the chamber exposure time present in the calibration table does not meet a preset standard, the autofocus exposure time and the chamber exposure time are based on an algorithm regarding the relationship between image intensity. At least one of the chamber exposure times may be updated.

In one embodiment, the fine particle counting device includes a slide guide unit including at least one wire hole; and a wire connected to the at least one wire hole for holding the slide, wherein the wire may be formed in a shape corresponding to the slide.

One embodiment of the present disclosure seeks to provide a method of operating a fine particle counting device. In one embodiment, the method of operating the fine particle counting device includes determining an index exposure time; determining an autofocus exposure time and a chamber exposure time corresponding to the index exposure time using a calibration table; Identifying the alignment portion and the insertion of the slide containing the chamber; performing an automatic focusing procedure of the alignment portion of the slide based on the autofocus exposure time determined using the calibration table; and acquiring an image of the chamber of the slide based on the chamber exposure time determined using the calibration table.

In one embodiment, determining the index exposure time includes identifying a target image intensity value; and determining an exposure time corresponding to the target image intensity value as an index exposure time before the slide is inserted.

In one embodiment, determining the index exposure time may include determining the index exposure time based on a binary search algorithm.

In one embodiment, the index exposure time, the autofocus exposure time, and the chamber exposure time may be mapped to each other and stored in the calibration table.

In one embodiment, the method includes identifying a target image intensity value; determining the index exposure time corresponding to the target image intensity value, and identifying that the autofocus exposure time and chamber exposure time corresponding to the index exposure time are not in the calibration table; determining a first autofocus exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the autofocus exposure time and image intensity; determining a first chamber exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the chamber exposure time and image intensity; and updating the calibration table by associating the index exposure time, the first autofocus exposure time, and the first chamber exposure time with the calibration table.

In one embodiment, when at least one of the autofocus exposure time and the chamber exposure time present in the calibration table does not meet a preset standard, the autofocus exposure time and the chamber exposure time are based on an algorithm regarding the relationship between image intensity. At least one of the chamber exposure times may be updated.

In one embodiment, the fine particle counting device includes a slide guide unit including at least one wire hole; and a wire connected to the at least one wire hole for holding the slide, wherein the wire may be formed in a shape corresponding to the slide.

One embodiment of the present disclosure includes a program stored in a recording medium to execute the method according to the embodiment of the present disclosure on a computer.

An embodiment of the present disclosure includes a computer-readable recording medium on which a program for executing a method according to an embodiment of the present disclosure on a computer is recorded.

An embodiment of the present disclosure includes a computer-readable recording medium that records a database used in an embodiment of the present disclosure.

According to an embodiment of the present disclosure, an automatic correction procedure for acquiring a specific target image value can be performed more quickly.

1 is a diagram showing a slide including an alignment unit and a chamber according to an embodiment of the present disclosure.
Figure 2 is a flowchart showing a method of operating a fine particle counting device according to an embodiment of the present disclosure.
Figure 3 is an example of a calibration table according to an embodiment of the present disclosure. The unit of each exposure time is microsecond (μs).
Figure 4 is a flowchart showing a method of updating a calibration table according to an embodiment of the present disclosure.
Figure 5 is a flowchart showing a method of updating a calibration table according to a chamber image acquisition result according to an embodiment of the present disclosure.
Figure 6 is a diagram showing a method of inserting a slide into a fine particle counting device according to an embodiment of the present disclosure.
Figure 7 is a diagram showing a slide receiving portion of a fine particle counting device according to an embodiment of the present disclosure.
Figure 8 is a diagram showing the shape of a slide accommodated in the slide receiving portion of a fine particle counting device according to an embodiment of the present disclosure.
Figure 9 is a block diagram showing a fine particle counting device according to an embodiment of the present disclosure.

In order to clarify the technical idea of the present disclosure, embodiments of the present disclosure will be described in detail with reference to the attached drawings. In describing the present disclosure, if it is determined that a detailed description of a related known function or component may unnecessarily obscure the gist of the present disclosure, the detailed description will be omitted. Components having substantially the same functional configuration among the drawings are given the same reference numbers and symbols as much as possible, even if they are shown in different drawings. For convenience of explanation, if necessary, the device and method should be described together. Each operation of the present disclosure does not necessarily have to be performed in the order described, and may be performed in parallel, selectively, or individually.

The terms used in the embodiments of the present disclosure have selected general terms that are currently widely used as much as possible while considering the function of the present disclosure, but this may vary depending on the intention or precedent of a person working in the art, the emergence of new technology, etc. . In addition, in certain cases, there are terms arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the relevant embodiment. Therefore, the terms used in this specification should not be defined simply as the names of the terms, but should be defined based on the meaning of the term and the overall content of the present disclosure.

Throughout this disclosure, singular expressions may include plural expressions, unless the context clearly dictates otherwise. Terms such as "include" or "have" are intended to designate the presence of a feature, number, step, operation, component, part, or combination thereof, but not one or more other features, numbers, steps, operations, or composition. It should be understood that this does not exclude in advance the possibility of the presence or addition of elements, parts, or combinations thereof. In other words, when it is said that a part "includes" a certain element throughout the present disclosure, this means that other elements may be further included rather than excluding other elements, unless specifically stated to the contrary.

An expression such as "at least one" modifies the entire list of elements, not the elements of the list individually. For example, “at least one of A, B, and C” and “at least one of A, B, or C” means only A, only B, only C, both A and B, both B and C, and A and C All refers to all of A, B, and C, or a combination thereof.

In addition, terms such as "...unit" and "...module" described in the present disclosure mean a unit that processes at least one function or operation, which is implemented in hardware or software or by a combination of hardware and software. It can be implemented.

Throughout the present disclosure, when a part is said to be “connected” to another part, this includes not only the case where it is “directly connected” but also the case where it is “electrically connected” with another element in between. do. Additionally, when a part "includes" a certain component, this means that it may further include other components rather than excluding other components, unless specifically stated to the contrary.

The expression “configured to” used throughout the present disclosure may be used, depending on the context, for example, “suitable for,” “having the capacity to.” )", "designed to", "adapted to", "made to", or "capable of". . The term “configured (or set to)” may not necessarily mean “specifically designed to” in hardware. Instead, in some contexts, the expression “system configured to” may mean that the system is “capable of” in conjunction with other devices or components. For example, the phrase "processor configured (or set) to perform A, B, and C" refers to a processor dedicated to performing the operations (e.g., an embedded processor), or by executing one or more software programs stored in memory. It may refer to a general-purpose processor (e.g., CPU or application processor) that can perform the corresponding operations.

Terms containing ordinal numbers, such as first, second, etc., may be used to describe various components, but the components are not limited by the terms. Terms are used only to distinguish one component from another. For example, a first component may be referred to as a second component, and similarly, the second component may be referred to as a first component without departing from the scope of the present disclosure. The term and/or includes a combination of a plurality of related items or any one item among a plurality of related items.

The term “about” used in the present disclosure may mean within 10% of a given value or range, preferably within 5%, and more preferably within 1%.

Throughout this disclosure, microparticles include cells, red blood cells, platelets, white blood cells, circulating tumor cells, stem cells, effete stored erythrocytes, T-cells derived from autologous T-cell expansion, organic microparticles, and inorganic microparticles. Microparticles, organometallic microparticles, metallic microparticles, aerosol particles, bacteria, yeast, fungi, algae, viruses, microinvertebrates or their eggs, pollen, cell or tissue fragments, cell masses, cell debris (e.g. DNA) or cellular debris associated with RNA purification), bioreactor-produced cells or granules, proteins, protein aggregates, prions, vesicles, liposomes, precipitates (e.g., precipitates from blood or blood fractions, industrial process precipitates, wastewater precipitates, etc.), It may include particles or cells from fermented foods (e.g., particles or cells from fermented beverages), macromolecules, macromolecular aggregates, DNA, cellular organelles, spores, vesicles, droplets, and exosomes.

1 is a diagram showing a slide including an alignment unit and a chamber according to an embodiment of the present disclosure.

Referring to FIG. 1, the slide 100 may be provided with one or more channels. Although the slide 100 is shown in FIG. 1 with four channels, this is only an example and the slide 100 may be provided with a variety of channels, such as 1, 2, 3, or 5 or more channels. Additionally, the slide 100 may include a plurality of alignment parts corresponding to each of a plurality of channels. In one embodiment, the alignment unit 110 may be arranged not to overlap the chamber 130. In one embodiment, the alignment unit 110 may include an align key in the form of a square grid, for example, a square grid. For example, the alignment unit 110 may have a square grid shape of approximately 40㎛.

In one embodiment, the slide 100 includes an alignment unit 110, an injection port 120 for injecting a sample, a chamber 130 for receiving the sample injected through the injection port, and a chamber for the sample to spread into the chamber 130. It may include an outlet 140 formed in the lower area of . The outlet 355 may be formed in an area opposite to the inlet 345. For example, the inlet 345 may be formed in the upper region of the chamber, and the outlet 355 may be formed in the lower region of the chamber. However, not all of the components shown in FIG. 1 are essential components of the slide 100. The slide 100 may be implemented with more components than those shown in FIG. 1, or the slide 100 may be implemented with fewer components than the components shown in FIG. 1. For example, the slide 100 may further include a channel identification unit that displays the identifier of the channel, an insertion direction display unit that indicates the insertion direction of the slide, and a sample capacity display unit that displays the injection volume of the sample.

In one embodiment, the slide 100 may be designed so that no air bubbles enter the chamber when microparticles are loaded into the channel.

According to one embodiment, the chamber 130 itself is provided on the slide 100, thereby eliminating the need for a cover glass, thereby reducing the risk of sample infection. Furthermore, more accurate volume control of the sample may be possible. . According to one embodiment, the volume of the chamber 130 can be formed to be constant by manufacturing the slide 100 with the chamber 130 instead of using a manual method of covering the slide glass with a cover glass. The sample capacity display unit may indicate the volume of the chamber 130. For example, if the volume of the chamber is about 20 μl, “20 μl” may be displayed on the sample capacity display.

According to one embodiment, by keeping the height of the chamber 130 of the slide 100 constant, the volume of the sample to be measured is maintained constant, thereby enabling accurate fine particle counting.

According to one embodiment, the chamber 130 itself is provided on the slide 100, thereby eliminating the need for a cover glass, thereby reducing the risk of sample infection. Furthermore, more accurate volume control of the sample may be possible. .

The slide 100 of FIG. 1 includes a plurality of alignment units corresponding to each of a plurality of channels, but may include only one alignment unit.

According to one embodiment, when the alignment unit is positioned so as not to overlap the chamber, the cell staining solution or cells mixed in the sample may not affect the grid shape of the alignment unit. Additionally, since the alignment unit is located outside the chamber, the fine particle counting device can count the center of the chamber, thereby increasing counting accuracy.

Figure 2 is a flowchart showing a method of operating a fine particle counting device according to an embodiment of the present disclosure.

Referring to FIG. 2, an image-based fine particle counting device can obtain reliable results only by counting fine particles using an image of a chamber with a certain intensity. It is possible to automatically adjust the brightness of the image through software processing, but the quality of the image processed through software may not be as good as the original image acquired through the camera. In addition, in the fine particle counting device, a dye (for example, Trypan blue) is used to facilitate the analysis of fine particles. Since the concentration of the dye may vary depending on the user, the image acquired through the camera An automatic brightness adjustment procedure is required. Factors that control the intensity of the image acquired through the camera include brightness, contrast, gain, and exposure time. In particular, the process of calculating the intensity of the image each time and adjusting the exposure time to match the target value requires many calculations. To quickly perform these calculations, most camera sensors provide an auto exposure function, but the auto exposure function is mainly optimized for everyday shooting environments and is suitable for industrial or research fields where uniform images must be obtained. You may not. Accordingly, a manual exposure control function, rather than automatic exposure, may be used. When using the manual exposure function, the image intensity reacts sensitively even to small exposure values, and it is necessary to finely adjust the exposure time several times to obtain the desired image intensity. Furthermore, in a fine particle counting device, a light source (e.g., LED) passes through the slide and is detected by the camera sensor. In this case, even if the light source is set to be the same, the camera image may change depending on the transmittance of the slide, LED difference, and camera sensor deviation. Strength values may vary from device to device. In other words, deviations can occur not only in different equipment, but also in the same equipment.

The present disclosure seeks to provide a fine particle counting device and an image correction method to solve these problems.

In step 210, the fine particle counting device may determine the index exposure time in a state where the slide is not inserted. The index exposure time may be an exposure time corresponding to a specific target image intensity value. In one embodiment, the fine particle counting device may determine the exposure time corresponding to the target image intensity value as the index exposure time using a binary search tree in the absence of a slide sample, for example, when the product is booted. . The index exposure time is a standard value depending on the status of the light source, camera, etc. of the current fine particle counting device, and varies depending on given conditions, so the fine particle counting device can determine the index exposure time every time it boots.

In one embodiment, the fine particle counting device may identify whether a value corresponding to the index exposure time determined in step 210 exists in the calibration table. If a value corresponding to the index exposure time exists in the calibration table, the fine particle counting device may perform step 230.

In step 230, the fine particle counting device may determine an autofocus exposure time and a chamber exposure time corresponding to the index exposure time using a calibration table. In one embodiment, the autofocus exposure time refers to the exposure time according to the target brightness value of the alignment unit 110 used for autofocus, and the chamber exposure time refers to the target value of the area of the chamber 130. It can refer to the exposure time according to .

In one embodiment, the index exposure time, autofocus exposure time, and chamber exposure time may be stored in correspondence with each other in the calibration table. For example, at least hundreds of index exposure times and autofocus exposure times and chamber exposure times corresponding to each of the index exposure times may be stored in the calibration table. Alternatively, a first calibration table in which index exposure time and autofocus exposure time correspond to each other and a second calibration table in which index exposure time and chamber exposure time correspond to each other may be used. Accordingly, the fine particle counting device can identify the autofocus exposure time and chamber exposure time corresponding to the index exposure time determined in step 210 on the calibration table.

In one embodiment, the slide used for counting fine particle counting devices has an alignment unit 110 and a chamber 130 separated as shown in FIG. 1, and the chamber 130, unlike the alignment unit 110, contains dye. An image of the chamber can be acquired in this state. Accordingly, both a brightness correction procedure to better obtain an image of the alignment unit 110 and a brightness correction procedure to better obtain an image of the chamber 130 may be necessary. That is, both the autofocus exposure time required for the brightness correction procedure to better acquire the image of the alignment unit 110 and the chamber exposure time required for the brightness correction procedure to better acquire the image of the chamber 130 may be required. .

In the case of a slide in which the alignment part overlaps the chamber, only the index exposure time and chamber exposure time are required, and the index exposure time and chamber exposure time are obtained using a binary search algorithm. However, according to one embodiment, the slide (100 ) is located in a state where the alignment unit and the chamber do not overlap each other, so it may take a lot of time to determine the index exposure time, autofocus exposure time, and chamber exposure time using a binary search algorithm. Therefore, the time required for image correction can be shortened by previously storing a calibration table that matches at least one of the index exposure time, autofocus exposure time, and chamber exposure time, and using the calibration table in which the fine particle counting device is stored. there is.

In one embodiment, there may be a plurality of calibration tables. The fine particle counting device may store a plurality of calibration tables in the database according to the target image intensity value. For example, correction tables for target image intensity values of 128, 129, 130, 131, 132, and 133, respectively, may be stored.

For example, the calibration table in FIG. 3 may represent a portion of the calibration table when the target image intensity value is 130. Accordingly, when the target image intensity value is 130 and the index exposure time is 191 μs (microsecond), the autofocus exposure time and chamber exposure time can be identified as 1520 and 959 μs, respectively. Hereinafter, unless there is a separate unit description for the exposure time, all references are to microseconds (μs).

In step 250, the microparticle counting device may identify that a slide including an alignment unit and a chamber has been inserted. In one embodiment, the slide may be the slide shown in FIG. 1, or the slide shown in FIG. 1 may have only one alignment portion.

In step 270, the fine particle counting device may perform an automatic focusing procedure of the alignment portion of the slide based on the autofocus exposure time determined using the calibration table. For example, if the calibration table with a target image intensity value of 130 is shown in Figure 3, and the fine particle counting device determines the index exposure time to be 22 μs, the fine particle counting device corresponds to the target image intensity value of 130. Based on the calibration table, the autofocus exposure time can be identified as 7158 μs. Accordingly, the fine particle counting device can acquire an image of the alignment part with an exposure time of 7158 μs.

In step 290, the fine particle counting device may acquire an image of the chamber of the slide based on the chamber exposure time determined using the calibration table. For example, if the calibration table with a target image intensity value of 130 is shown in Figure 3, and the fine particle counting device determines the index exposure time to be 22 μs, the fine particle counting device corresponds to the target image intensity value of 130. Based on the calibration table, the chamber exposure time can be identified as 23999 μs. Accordingly, the fine particle counting device can acquire an image of the chamber with an exposure time of 23999 μs.

According to one embodiment, calibration tables are stored in the database so that two or more of the index exposure time, autofocus exposure time, and chamber exposure time are stored in association, so when the fine particle counting device determines the index exposure time, Autofocus exposure times and chamber exposure times can also be identified directly via the calibration table. Since the image to be ultimately acquired using the fine particle counting device is a chamber image, the operating time of the fine particle counting device can be shortened by minimizing the time to acquire the chamber exposure time using a calibration table. If the index exposure time determined by the fine particle counting device does not exist on the calibration table, this will be described in more detail later with reference to FIG. 4.

Meanwhile, although FIG. 2 illustrates a method of operating a fine particle counting device according to an embodiment of the present disclosure, various modifications may be made to FIG. 2 of course. As an example, although sequential operations are shown in FIG. 2, various operations in FIG. 2 may overlap, be performed in parallel, may be performed in a different order, or some operations may be performed multiple times repeatedly. Of course it can be done.

Figure 3 is an example of a calibration table according to an embodiment of the present disclosure.

Referring to FIG. 3, a portion of the calibration table when the target image intensity value is 130 is shown as an example. A plurality of lists may be stored in the calibration table, and a plurality of calibration tables may be stored in the database according to the target image intensity value. That is, each calibration table may exist depending on the target image intensity value.

Figure 4 is a flowchart showing a method of updating a calibration table according to an embodiment of the present disclosure.

Referring to FIG. 4, in step 410, the fine particle counting device may identify the target image intensity value. In one embodiment, an image-based fine particle counting device can perform reliable counting only by acquiring an image with a certain image intensity, thereby identifying a target image intensity that can facilitate analysis. The target image intensity may be input through user input, or may be determined by a fine particle counting device in consideration of the shooting environment. In one embodiment, the target image intensity value may include a target image intensity value for determining an index exposure time, a target image intensity value for determining an autofocus exposure time, and a target image intensity value for determining a chamber exposure time. there is. Target image intensity values for determining the index exposure time, autofocus exposure time, and chamber exposure time may mean the same value, may have different values, or at least some of them may have the same value.

In step 420, the fine particle counting device may determine an index exposure time corresponding to the target image intensity value. In one embodiment, the fine particle counting device may determine an index exposure time corresponding to the target image intensity value in the absence of a slide. For example, a particle counting device may determine an index exposure time corresponding to a target image intensity value each time the device is booted.

In one embodiment, the fine particle counting device may determine the index exposure time corresponding to the target image intensity value based on a binary search algorithm. For example, using a binary search algorithm, if the target image intensity value is 130, the fine particle counting device identifies the image intensity as 30 when the exposure time is 100, and the image intensity when the exposure time is 3000. We can identify the intensity as 226, and based on 130 being between 30 and 226, we can identify the image intensity when the exposure time is 1550. Based on the fact that the image intensity is 170 when the exposure time is 1550, the fine particle counting device uses the image intensity when the exposure time is 800, which is about the middle value between 1550 and 30, to find the exposure time when the image intensity is 130. Based on the fact that the image intensity when the exposure time is 800 is less than 130, the image intensity at 1200, which is approximately between 800 and 1550, can be detected. In this way, using binary search, the exposure time with an image intensity of 130 can be determined as 1250, and the exposure time with an image intensity of 130, 1250, can be determined as the index exposure time. The index exposure time determined in this way can serve as a kind of control for the autofocus exposure time and chamber exposure time to be determined later.

In step 430, the fine particle counting device may identify whether the determined index exposure time value exists in the calibration table. In one embodiment, the calibration table may be information that has already been created and stored in a database. The fine particle counting device may perform step 440 if the determined index exposure time value is identified as a value that exists in the calibration table, and may perform step 450 if it is identified as not existing.

In step 440, the fine particle counting device may determine the autofocus exposure time and chamber exposure time corresponding to the index exposure time using a calibration table. In one embodiment, step 440 may correspond to step 230 described above with reference to FIG. 2. Accordingly, step 250 may be performed after step 440.

Hereinafter, a case where it is identified that there is no index exposure time value in the calibration table as shown in FIG. 3 will be described as an example.

At step 450, the particle counting device, based on identifying that the index exposure time value is not in the calibration table, generates a first automatic signal corresponding to the target image intensity based on an algorithm regarding the relationship between autofocus exposure time and image intensity. You can determine the focus exposure time. For example, an algorithm regarding the relationship between autofocus exposure time and image intensity may be a binary search algorithm.

For example, if the target image intensity for determining the autofocus exposure time is 130, the fine particle counting device identifies the image intensity as 120 when the autofocus exposure time is set to 3400, and the autofocus exposure time is set to 3600. Based on identifying the image intensity as 140, the image intensity when the intermediate autofocus exposure time is set to 3400 can be identified as 138. Accordingly, the fine particle counting device can search for an autofocus exposure time with an image intensity of 130 by setting a value between 3400 and 3600 as the autofocus exposure time. If the fine particle counting device identifies the first autofocus exposure time with an image intensity of 130 as 3526, the autofocus exposure time may be set to 3526 to photograph the slide alignment portion to perform an autofocusing procedure.

In step 460, the fine particle counting device may determine the first chamber exposure time corresponding to the target image intensity based on an algorithm regarding the relationship between chamber exposure time and image intensity. For example, an algorithm regarding the relationship between autofocus exposure time and image intensity may be a binary search algorithm.

For example, if the target image intensity for determining the chamber exposure time is 130, the fine particle counting device identifies the image intensity when the chamber exposure time is 3000 as 122, and when the chamber exposure time is 3200, the fine particle counting device identifies the image intensity as 122. Based on identifying the image intensity as 136, the image intensity when the intermediate chamber exposure time is set to 3100 can be identified as 134. Accordingly, the fine particle counting device can search for a chamber exposure time with an image intensity of 130 by setting a value between 3000 and 3200 as the chamber exposure time. When the fine particle counting device identifies the exposure time of the first chamber with an image intensity of 130 as 3012, the chamber image can be obtained by photographing the chamber of the slide by setting the chamber exposure time to 3012.

In one embodiment, when the fine particle counting device determines a first chamber exposure time having a target image intensity, the fine particle counting device may acquire an image of the chamber using the first chamber exposure time.

In step 470, the fine particle counting device may update the calibration table corresponding to a specific image intensity by associating the index exposure time, the first autofocus exposure time, and the first chamber exposure time with the calibration table. For example, based on determining that the index exposure time corresponding to the target image intensity 130 is 1250, the first autofocus exposure time is 3526, and the second chamber exposure time is 3012, the calibration table includes index exposure time 1250 and autofocus. Exposure time 3526 and chamber exposure time 3012 may be stored in association.

Meanwhile, although FIG. 4 illustrates a method of updating a calibration table according to an embodiment of the present disclosure, various modifications may be made to FIG. 4 of course. As an example, although sequential operations are shown in FIG. 4, various operations in FIG. 4 may overlap, may be performed in parallel, may be performed in a different order, or some operations may be performed multiple times repeatedly. Of course it can be done.

The image correction method according to an embodiment of the present disclosure can be applied to all cameras. Two types of cameras are usually used in fine particle counter equipment: one is a global shutter type that acquires the entire image at once, and the other is a method that acquires the image line by line rather than as a complete frame. It is a rolling shutter method obtained. Global shutter cameras have the advantage of having a relatively short delay time in image acquisition, but are expensive. On the other hand, in the case of rolling shutter, delay time is required to obtain an accurate image, but since it is low cost, the price is competitive. Embodiments of the present disclosure provide an image correction method that can be applied to both types of cameras.

In addition, the higher the fps (frame per second; that is, the screen conversion rate per second), the better, but in terms of price competitiveness, embedded systems with low fps (frame per second) are often used.

When using the calibration table according to an embodiment of the present disclosure, even when using a rolling shutter camera or equipment with a low fps, equipment for obtaining a desired target image intensity value by minimizing the automatic image adjustment time of the camera The operating time can be shortened. For example, even if the fps is high, most low-cost cameras use a rolling shutter, so a delay time is provided to obtain an accurate image. When using the calibration table according to an embodiment of the present disclosure, the automatic calibration procedure is performed quickly. This can be done and the time can be shortened.

Figure 5 is a flowchart showing a method of updating a calibration table according to a chamber image acquisition result according to an embodiment of the present disclosure.

Referring to FIG. 5, when the intensity value of the image of the chamber is not the target image intensity value as a result of acquiring the image of the chamber based on the chamber exposure time existing in the calibration table, the process of updating the calibration table is shown. do.

In step 500, the fine particle counting device may determine an index exposure time corresponding to the target image intensity value. In one embodiment, step 500 may correspond to step 210 described above with reference to FIG. 2 or step 420 described above with reference to FIG. 4.

In one embodiment, the fine particle counting device may identify whether a value corresponding to the index exposure time determined in step 500 exists in the calibration table. If a value corresponding to the index exposure time exists in the calibration table, the fine particle counting device may perform step 510.

In step 510, the fine particle counting device may determine the first autofocus exposure time and the first chamber exposure time corresponding to the index exposure time using a calibration table. The first autofocus exposure time and the first chamber exposure time may be values stored in the calibration table to correspond to the index exposure time.

In step 520, the fine particle counting device may acquire an image of the alignment portion using the first autofocus exposure time.

In step 530, the fine particle counting device may determine whether the image of the alignment unit satisfies a preset first standard. For example, the fine particle counting device may measure the intensity of the obtained image of the alignment portion and determine whether the intensity of the image of the alignment portion is within a specific range. If the image intensity of the alignment is sufficient to perform an automatic focusing procedure, the particle counting device may perform step 560, otherwise, step 540 may be performed.

At step 540, the particle counting device may determine a second autofocus exposure time based on an algorithm regarding the relationship between autofocus exposure time and image intensity. For example, an algorithm regarding the relationship between autofocus exposure time and image intensity may be a binary search algorithm. Accordingly, a second autofocus exposure time with an image intensity suitable for performing an autofocusing procedure can be determined. However, in some cases, the step (modification 520) of acquiring an image of the alignment unit using the second autofocus exposure time may be additionally performed (not shown), and in this case, steps after 520 are repeated as is to obtain the alignment unit image. The process of re-obtaining a third, fourth, or more autofocus exposure time may be repeated until the image satisfies the first preset criterion (not shown).

At step 550, the particle counting device may update the calibration table based on the second autofocus exposure time. In one embodiment, the fine particle counting device may update the calibration table by changing and storing the autofocus exposure time corresponding to the index exposure time from the first autofocus exposure time to the second autofocus exposure time.

At step 555, the particle counting device may perform an auto-focusing procedure based on the second auto-focus exposure time. For example, if the fine particle counting device determines in step 530 that the alignment unit image satisfies the first preset standard, the fine particle counting device may perform an automatic focusing procedure using the first autofocus exposure time. Alternatively, the fine particle counting device may update the calibration table based on the second autofocus exposure time in step 550 and then perform an automatic focusing procedure based on the second autofocus exposure time. In one embodiment, the calibration table update procedure of step 550 may be performed after the automatic focusing procedure of step 555.

In step 560, the fine particle counting device may acquire a chamber image using the first chamber exposure time. For example, the fine particle counting device may acquire a chamber image using the first chamber exposure time after focusing based on the result of automatic focusing.

In step 570, the fine particle counting device may determine whether the image of the chamber satisfies a preset second standard. For example, a fine particle counting device can measure the intensity of an acquired image of a chamber and determine whether the intensity of the image of the chamber is within a specific range. If the image intensity of the chamber is sufficient to perform particle counting, the particle counting device may perform step 595, otherwise, step 580 may be performed.

At step 580, the particle counting device may determine the second chamber exposure time based on an algorithm regarding the relationship between chamber exposure time and image intensity. For example, an algorithm relating the relationship between chamber exposure time and image intensity may be a binary search algorithm. Accordingly, a second chamber exposure time having an image intensity suitable for performing fine particle counting can be determined. However, in some cases, a step (modification 560) of acquiring a chamber image using the second chamber focus exposure time may be additionally performed (not shown), and in this case, steps after 560 are repeated as is, so that the chamber image is obtained. The process of re-obtaining a third, fourth, or more chamber focus exposure time may be repeated until the first preset criterion is satisfied (not shown).

At step 590, the particle counting device may update the calibration table based on the second chamber exposure time. In one embodiment, the fine particle counting device may update the calibration table by changing and storing the chamber exposure time corresponding to the index exposure time from the first chamber exposure time to the second chamber exposure time. Additionally, the fine particle counting device may acquire a chamber image based on the second chamber exposure time.

In step 595, the fine particle counting device may perform fine particle counting based on the chamber image acquired in step 560 or step 590.

Meanwhile, although FIG. 5 illustrates a method of updating a calibration table according to an embodiment of the present disclosure, various modifications may be made to FIG. 5 of course. As an example, although sequential operations are shown in FIG. 5, various operations in FIG. 5 may overlap, be performed in parallel, may be performed in a different order, or some operations may be performed multiple times repeatedly. Of course it can be done. For example, steps 530 to 550 may be omitted.

According to one embodiment, if the exposure time existing in the calibration table is not an optimal value, the calibration table is updated, and the optimal values can be used thereafter.

Figure 6 is a diagram showing a method of inserting a slide into a fine particle counting device according to an embodiment of the present disclosure.

Referring to FIG. 6, the fine particle counting device 600 may include a display 610, a slide receiving unit 630, and the like. In one embodiment, the fine particle counting device 600 may identify that the slide 620 is inserted through the slide receiving portion 630. For example, the fine particle counting device 600 automatically identifies that the slide 620 is inserted into the slide receiving unit 630, or obtains a user input that the slide 620 has been inserted and confirms that the slide 620 is inserted. can be identified.

Here, the slide 620 includes an alignment part, an inlet, an outlet, a chamber, etc., and the alignment part may be positioned so as not to overlap the chamber. For example, slide 620 may correspond to the slide described above with reference to FIG. 1 .

In one embodiment, the fine particle counting device 600 can identify whether the slide 620 is inserted in the forward direction. If the slide 620 is not inserted in the correct direction, the fine particle counting device 600 may display a warning message indicating that the insertion direction is incorrect.

In one embodiment, the fine particle counting device 600 may obtain user input for the injection volume of the sample, and obtain information on fine particles located in the chamber of the slide 620 based on the user input for the injection volume. there is. Obtaining fine particle information may include counting fine particles. The slide 620 may further include a sample capacity display unit that displays the sample injection capacity so that the user can easily know the sample injection capacity.

In one embodiment, the fine particle counting device 600 may perform automatic focusing based on the alignment portion of the inserted slide 620. Automatic focusing may refer to a process in which a camera focuses to photograph the slide 620 before acquiring an image of the slide 620. In particular, since the fine particle counting device 600 is a device that observes very small fine particles, the lens focusing position can greatly affect image analysis. If the slide 620 is tilted or twisted for any reason, focusing for each shooting position must be performed again each time, which may delay the fine particle counting process. Accordingly, the slide receiving portion 630 of the fine particle counting device 600 may include a slide guide portion and a wire that help the slide be mounted consistently. This will be described in detail later with reference to FIGS. 7 and 8.

In one embodiment, the fine particle counting device 600 may acquire a chamber image of the slide 620 based on an automatic focusing result and obtain information on fine particles located in the chamber based on the chamber image. For example, fine particle information may include information on the number of fine particles located within a certain area of the chamber.

For example, when the slide 620 includes a plurality of alignment units corresponding to each of a plurality of chambers, the fine particle counting device 600 may detect each of the plurality of alignment units based on the automatic focusing results for each of the plurality of alignment units. Information on fine particles located in the chamber corresponding to can be obtained.

For another example, when the slide 620 includes a plurality of chambers and one alignment unit, the fine particle counting device 600 counts the fine particles located in the plurality of chambers based on the automatic focusing result for one alignment unit. Information can be obtained.

Figure 7 is a diagram showing a slide receiving portion of a fine particle counting device according to an embodiment of the present disclosure.

In one embodiment, the fine particle counting device is equipment that photographs fine particles present within a certain area of the chamber and obtains information about the fine particles. The small particles are small, so the focusing position of the lens has a great influence on image analysis. It can go crazy. If the slide 750 is accommodated in the slide receiving portion 630 in an inclined or twisted state, it may take some time to count fine particles because focusing must be performed for each shooting position each time. Additionally, if the mechanism that ensures that the slide 750 is properly accommodated in the slide receiving portion 630 is large, it may obscure the upper part of the thin slide, thereby limiting the imaging area. In addition, it is desirable that the mechanism that allows the slide 750 to be properly accommodated in the slide receiving portion 630 is designed to accommodate slides of various thicknesses. If the mechanism is made of a material with weak durability, Long-term use may not be possible due to wear due to friction. In addition, there is a problem in that the manufacturing cost increases when the relevant device has a complex shape or the manufacturing method is difficult.

One embodiment of the present disclosure seeks to provide a mechanism for properly accommodating the slide 750 in the slide receiving portion 630 to solve these problems.

Referring to FIG. 7, the slide accommodating part 630 for accommodating a slide includes at least one wire fixing part 730, a slide guide part 710, 720 including at least one wire hole 760, and at least one It may include a wire 740 for holding the slide, which is connected to the wire hole 760 of . The shape of the wire 740 may be manufactured to correspond to the slide. In one embodiment, the wire 740 may be made of stainless steel.

Figure 8 is a diagram showing the shape of a slide accommodated in the slide receiving portion of a fine particle counting device according to an embodiment of the present disclosure.

FIG. 8 shows a state in which the slide 750 is accommodated in the slide receiving portion 630 of FIG. 7 . The slide 750 can be accommodated in an appropriate position of the slide receiving portion 630 by the guide portions 710 and 720 and the wire 740. According to one embodiment, the wire 740 can physically bring the slide 750 into close contact with a flat surface to the extent that there is no problem with focusing the lens. Accordingly, the same or close focusing position can be provided each time the slide 750 is inserted. In particular, by using the wire 740 with a small wire diameter, the area covering the upper part of the slide 750 can be minimized. Accordingly, there can be greater freedom from constraints on the size and location of the chamber. Additionally, by using the wire 740 structure, the slide 750 of various thicknesses can be used. For example, slides having various thicknesses of 4 mm or less can be used.

In one embodiment, the wire 740 may be made of stainless steel. In this case, since the wire 740 has a higher hardness than the slide 750, the wire 740 may not be damaged due to friction with the slide 750 even if it is repeatedly used for a long period of time. In addition, complex shapes can be manufactured at a low cost through the wire forming process, and mass production can be possible. In addition, even if the design changes, it can be easily changed without affecting the unit cost. For example, the wire 740 may be transformed into various shapes other than those shown in Figures 7 or 8.

Figure 9 is a block diagram showing a fine particle counting device according to an embodiment of the present disclosure.

Referring to FIG. 9, the fine particle counting device 900 may include a memory 910, a display 920, a camera 930, and a processor 940. However, not all of the components shown in FIG. 9 are essential components of the fine particle counting device 900. The fine particle counting device 900 may be implemented with more components than those shown in FIG. 9, or the fine particle counting device 900 may be implemented with fewer components than the components shown in FIG. 9. there is. For example, the fine particle counting device 900 may further include a transmitter and receiver.

Various types of data, such as programs and files such as applications, can be installed and stored in the memory 910. The processor 940 may access and use data stored in the memory 910, or may store new data in the memory 910. For example, the memory 910 may store the focusing position of the lens. In one embodiment, memory 910 may include a database. Calibration tables corresponding to each image intensity value may be stored in the database.

The processor 940 controls the overall operation of the fine particle counting device 900 and may include at least one processor such as a CPU or GPU. The processor 940 may control other components included in the fine particle counting device 900 to perform operations for operating the fine particle counting device 900. For example, the processor 940 may execute a program stored in the memory 910, read a stored file, or save a new file. In one embodiment, the processor 940 may perform an operation to operate the fine particle counting device 900 by executing a program stored in the memory 910. For example, the processor identifies that a slide is inserted into the slide receiving unit, performs automatic focusing based on the alignment of the slide, acquires a slide chamber image using a camera based on the automatic focusing result, and acquires an image of the slide chamber. Based on the chamber image, information on fine particles located in the chamber can be obtained.

In one embodiment, the display 920 may display and output information processed by the fine particle counting device 900. For example, the display 920 may output fine particle counting results.

An embodiment of the present disclosure may also be implemented in the form of a recording medium containing instructions executable by a computer, such as program modules executed by a computer. Computer-readable media can be any available media that can be accessed by a computer and includes both volatile and non-volatile media, removable and non-removable media. Additionally, computer-readable media may include both computer storage media and communication media. Computer storage media includes both volatile and non-volatile, removable and non-removable media implemented in any method or technology for storage of information such as computer-readable instructions, data structures, program modules or other data. Communication media typically includes computer-readable instructions, data structures, or program modules and includes any information delivery medium.

The foregoing description of the present disclosure is for illustrative purposes, and a person skilled in the art to which the present disclosure pertains will understand that the present invention can be easily modified into other specific forms without changing its technical idea or essential features. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive. For example, each component described as unitary may be implemented in a distributed manner, and similarly, components described as distributed may also be implemented in a combined form.

The scope of the present disclosure is indicated by the claims described below rather than the detailed description above, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be construed as being included in the scope of the present disclosure. do.

Claims (15)

In the fine particle counting device,
a memory storing one or more instructions; and
At least one processor executing the one or more instructions stored in the memory,
The at least one processor executes the one or more instructions,
Determine the index exposure time,
If a value corresponding to the index exposure time exists in the calibration table, determine the autofocus exposure time and chamber exposure time corresponding to the index exposure time using the calibration table,
Identify the insertion of the slide containing the alignment portion and chamber;
Based on the autofocus exposure time determined using the calibration table, perform an automatic focusing procedure on the alignment portion of the slide,
A fine particle counting device that acquires an image of the chamber of the slide based on the chamber exposure time determined using the calibration table. The method of claim 1, wherein the at least one processor:
Identify the target image intensity value,
A fine particle counting device that determines an exposure time corresponding to the target image intensity value as an index exposure time before the slide is inserted. The method of claim 1, wherein the at least one processor:
A fine particle counting device that determines the index exposure time based on a binary search algorithm. The method of claim 1, wherein the calibration table includes:
A fine particle counting device wherein the index exposure time, the autofocus exposure time, and the chamber exposure time are mapped to each other and stored. The method of claim 2, wherein the at least one processor:
Identifying that a value corresponding to the index exposure time is not in the calibration table,
determine a first autofocus exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the autofocus exposure time and image intensity;
determine a first chamber exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the chamber exposure time and image intensity;
updating the calibration table by associating the index exposure time, the first autofocus exposure time, and the first chamber exposure time with the calibration table. According to paragraph 1,
If at least one of the autofocus exposure time and the chamber exposure time present in the calibration table does not satisfy a preset standard, at least one of the autofocus exposure time and the chamber exposure time is based on an algorithm regarding the relationship between image intensities. One updated, fine particle counting device. The method of claim 1, wherein the fine particle counting device,
A slide guide unit including at least one wire hole; and
Further comprising a wire for holding the slide, connected to the at least one wire hole,
The wire is,
A fine particle counting device formed in a shape corresponding to the slide. As a method of operating a fine particle counting device,
determining an index exposure time;
determining an autofocus exposure time and a chamber exposure time corresponding to the index exposure time using a calibration table;
Identifying the alignment portion and the insertion of the slide containing the chamber;
performing an automatic focusing procedure of the alignment portion of the slide based on the autofocus exposure time determined using the calibration table; and
Acquiring an image of the chamber of the slide based on a chamber exposure time determined using the calibration table. The method of claim 8, wherein determining the index exposure time comprises:
identifying a target image intensity value; and
A method comprising determining an exposure time corresponding to the target image intensity value as an index exposure time before the slide is inserted. The method of claim 8, wherein determining the index exposure time comprises:
A method comprising determining the index exposure time based on a binary search algorithm. The method of claim 8, wherein the calibration table includes:
The method wherein the index exposure time, the autofocus exposure time, and the chamber exposure time are mapped to each other and stored. The method of claim 8, wherein
identifying a target image intensity value;
Determine the index exposure time corresponding to the target image intensity value,
identifying that the autofocus exposure time and chamber exposure time corresponding to the index exposure time are not in the calibration table;
determining a first autofocus exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the autofocus exposure time and image intensity;
determining a first chamber exposure time corresponding to the target image intensity value based on an algorithm regarding the relationship between the chamber exposure time and image intensity; and
updating the calibration table by correlating the index exposure time, the first autofocus exposure time, and the first chamber exposure time to the calibration table. According to clause 8,
If at least one of the autofocus exposure time and the chamber exposure time present in the calibration table does not satisfy a preset standard, at least one of the autofocus exposure time and the chamber exposure time is based on an algorithm regarding the relationship between image intensities. One is updated, how. The method of claim 8, wherein the fine particle counting device,
A slide guide unit including at least one wire hole; and
Further comprising a wire for holding the slide, connected to the at least one wire hole,
The wire is,
A method formed in a shape corresponding to the slide. A program stored in a computer-readable recording medium to execute the method of any one of claims 8 to 14 on a computer.

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