TWI717910B - Method of calculating cell volume - Google Patents

Abstract

The present invention provides a method for calculating cell volume. The steps include scanning a detection carrier and obtaining a three-dimensional image. The detection carrier is provided with at least one cell, and then obtaining a plurality of planes of the three-dimensional image according to the plurality of depth values of the three-dimensional image Image, the planar images include the at least one cell, obtain at least one cell image each included in the planar images by an edge retouching method, and calculate the area to obtain a plurality of area values, and finally superimpose the In the planar image, the volume of the at least one cell included in the detection carrier is calculated by using the depth values and the at least one cell image individually included in the planar images.

Description

Method of calculating cell volume

The present invention relates to a method for calculating cell volume, in particular to a method that uses a plurality of depth values to correspond to a plurality of planar images and a plurality of cell images, and superimposes the planar images, and calculates the cell volume with the cell images的方法。 The method.

The evolution of the times has led to the development of biological analysis, especially cell counting and measurement technology, which has increased the quantity and quality of information obtained from biological samples, and the medical community that uses it as a diagnostic tool has also benefited.

Known cell counting and measurement techniques include a biological technique, flow cytometry. Flow cytometry has applications in many fields, including molecular biology, pathology, immunology, and botany , And marine biology, etc. In molecular biology, it is mainly used for fluorescently labeled antibodies. Specific antibodies bind to antigens on target cells, and the special information of these cells can be studied by flow cytometry, which has a wide range of applications in medicine.

In flow cytometry, a beam of monochromatic light (usually a laser) shines on a stream of fluid that is hydrodynamically focused. Several detectors aim at the point where the beam and the laser intersect, one of which is on the same line as the laser (called forward scatter (FSC)), and the others are perpendicular to the laser (side scatter (SSC) and one Or several fluorescent monitors). When each suspended particle passes through the light beam, it scatters the light in a certain way, and at the same time the fluorescent compound it carries is excited and emits fluorescence with a lower frequency than the excitation light; the combined data of these scattered light and fluorescence is detected by the detector Record, according to the fluctuation of the brightness of each detector (each cell will show a peak of scattering or fluorescence), the physical and chemical properties of each particle can be calculated. Forward scatter is related to cell volume, while side scatter depends on the internal complexity of the particles (such as the shape of the nucleus, the type of particles in the cytoplasm or the roughness of the membrane). As for fluorescence, it is used to label and detect specific cell surface antigens or DNA.

However, the conventional flow cytometry technology still has shortcomings. One of the shortcomings is that the tubing used in the flow cytometry technology and the flow rate controller used to control the flow of the diluted blood sample through the sensor are different from the conventional flow cytometry technology. The pipeline in the blood cell technology device often leaks, which affects the accuracy and safety of the equipment. Another disadvantage of the conventional flow blood cell device is the accuracy of the internal flow controller and automatic dilution equipment. The flow cytometer The accuracy depends on the accuracy of the flow controller and sample dilution equipment, as well as the ability to maintain accurate calibration. The flow controller and dilution equipment often need to be recalibrated regularly, and the need to recalibrate means that the existing flow cytometry device has potential The possibilities and undesired operating costs are published in Cytometry Supplement.

In view of the problems of the above-mentioned conventional technology, the present invention provides a method for calculating cell volume, which scans the detection carrier and obtains its three-dimensional image. After obtaining a plurality of plane images of the three-dimensional image according to different depth values, the image is modified with edges Method to obtain the cell images contained in the plane images, calculate the area, and finally superimpose the plane images, use the depth values and the cell images to calculate the cell volume of the detection carrier, and it can also be changed The scanning range is used to simultaneously calculate the number and total volume of multiple cells in the detection carrier.

One object of the present invention is to provide a method for calculating cell volume, which scans a detection carrier and obtains a three-dimensional image, then obtains a plurality of plane images of the three-dimensional image according to a plurality of depth values, and obtains the planes by edge modification The area of at least one cell image included in the image is calculated, and the plane images are finally superimposed, and the depth values and at least one cell image included in the plane images are used to calculate the number of at least one cell included in the detection carrier volume.

In order to achieve the aforementioned objectives and effects, the present invention provides a method for calculating the volume of cells. The steps include scanning a detection carrier to obtain a three-dimensional image. The detection carrier is provided with at least one cell; and according to the plurality of depths of the three-dimensional image Obtain a plurality of plane images of the three-dimensional image; perform an edge modification method on each of the plane images to obtain at least one cell plane image contained in each of the plane images; calculate each of the plane images Obtain a plurality of area values for an area of the at least one cell plane image included; superimpose the plane images, and calculate a volume of the at least one cell based on the depth values and the area values; using These steps provide for separating each cell from the three-dimensional image and calculating the volume of each cell.

In an embodiment of the present invention, the edge retouching method uses an edge detection method.

In an embodiment of the present invention, wherein the detection carrier is scanned to obtain a three-dimensional image, the step of arranging at least one cell on the detection carrier is to obtain the three-dimensional image by optical coherence tomography (OCT).

In an embodiment of the present invention, after performing an edge retouching method on each of the plane images, at least one cell plane image included in the plane images is obtained, and the plane images respectively include In the step where the at least one cell plane image corresponds to one of the depth values, the at least one cell plane image corresponds to at least one cell.

In an embodiment of the present invention, in the step of superimposing the planar images, and calculating the volume of the at least one cell according to the depth values and the area values, the depth values correspond to them The area values are multiplied to calculate the volume of the at least one cell.

In an embodiment of the present invention, the at least one cell line is one cell colony or multiple cell colonies.

In an embodiment of the present invention, in the step of obtaining a plurality of plane images of the 3D image according to the plurality of depth values of the 3D image, the plane images include the at least one cell, the depth values are the The depth value of at least one cell in the three-dimensional image.

In order to enable your reviewer to have a further understanding and understanding of the features of the present invention and the effects achieved, the following examples and accompanying descriptions are provided. The description is as follows:

The present invention provides a method for calculating cell volume. The steps include scanning a detection carrier and obtaining a three-dimensional image. The detection carrier is provided with at least one cell, and then obtaining a plurality of planes of the three-dimensional image according to the plurality of depth values of the three-dimensional image Image, the planar images include the at least one cell, obtain at least one cell image each included in the planar images by an edge retouching method, and calculate the area to obtain a plurality of area values, and finally superimpose the In a planar image, the at least one cell volume included in the detection carrier is calculated using the depth values and the at least one cell image separately included in the planar images, and the steps are used to provide separation of each cell from the three-dimensional image, And calculate the volume of each cell method.

Please refer to Figure 1, which is a schematic diagram of the structure of an embodiment of the present invention. As shown in the figure, it is a method for calculating cell volume 1, which can calculate the volume of cells and the number of plural cells. The steps include:

Step S2: Scan a detection carrier to obtain a three-dimensional image, and the detection carrier is provided with at least one cell;

Step S4: Obtain a plurality of plane images of the 3D image according to the plurality of depth values of the 3D image, the plane images including at least one cell;

Step S6: After performing edge modification on each of the planar images, obtain at least one cell planar image included in each of the planar images;

Step S8: Calculate the area of at least one cell plane image included in each of the plane images to obtain a plurality of area values; and

Step S10: superimpose the planar images, and calculate the volume of at least one cell according to the depth values and the area values.

Please refer to Figure 1 again and refer to Figures 2A to 2D. Figures 2A to 2D are schematic diagrams of steps of an embodiment of the present invention. As shown in the figure, in step S2, one of the scanning devices is used to scan the lens 1 emit a scanning light 2 to scan a detection carrier 10 with a scanning range 3 to obtain a three-dimensional image 20. The inner side of the detection carrier 10 contains at least one cell 12; in this embodiment, the scanning device uses optical coherence tomography Scanning method (Optical coherence tomography, OCT) to obtain the three-dimensional image 20, the optical coherence tomography system can obtain transparent or opaque material surface and subsurface images, which uses an interferometer and a low-coherence broadband light source. The light is divided into two beams, namely the reference light and the detection light, and the technique of combining the reference light and the detection light to generate an interference pattern; in this embodiment, the size of the scanning range 3 can be determined according to the needs of the user (eg 3×3 mm ² ), this embodiment is not limited here; in this embodiment, the detection carrier 10 is used to carry the at least one cell 12, such as a porous microplate (or a porous culture plate) for culturing cells In this embodiment, the at least one cell 12 may be a single cell, a colony composed of multiple cells, or a plurality of cell colonies; the three-dimensional image 20 is a three-dimensional image of the detection carrier 10, which contains the at least one cell 12 Three-dimensional images.

Following the above, in step S4, as shown in Figure 2B, a plurality of planar images 30 of the 3D image 20 are obtained according to the plurality of depth values 22 of the 3D image 20, and the planar images 30 include the at least one The cell 12, that is, when the three-dimensional images 20 are cut by the planar images 30, the at least one cell 12 must be cut so that the planar images 30 each include the cut surface of the at least one cell 12; in this embodiment In this embodiment, each of the planar images 30 corresponds to a different depth, that is, the planar images 30 correspond to one of the depth values 22; in this embodiment, each of the planar images 30 The planar images 30 are parallel to each other; in this embodiment, the depth values 22 are the depth values of the at least one cell 12 in the three-dimensional image 20, that is, the depth values 22 correspond to the planar images 30. The depth value of the cut surface of the at least one cell 12 not included.

Following the above, in step S6, in this embodiment, an edge modification method is performed on each of the planar images 30 to find the cut surface of the at least one cell 12 contained in the planar images 30. At the edge, at least one cell plane image 122 included in each of the plane images 30 is obtained, wherein the at least one cell plane image 122 can be a single cell, a colony composed of multiple cells, or a planar image of a plurality of cell colonies; The edge retouching method of the embodiment uses an edge detection method, which can be the morphological gradient method in the edge detection method, but this embodiment is not limited here, and other edge detection methods can also be used.

Following the above, in step S8, in this embodiment, according to the scanning range 3 of the scanning lens 1, it can be known that the area of the unit pixel on each planar image 30 can be calculated. After digitizing an area of the at least one cell plane image 122 included in each of the plane images 30, a plurality of area values of the at least one cell plane image 122 included in the plane images 30 are obtained.

Following the above, in step S10, in this embodiment, as shown in Figure 2C and Figure 2D, the planar images 30 are sequentially superimposed, based on the depth values 22 and the cell planes. The area values of the image 122 are multiplied by the depth value 22 corresponding to the planar images 30 and the cell planar image 122 to calculate a volume of the at least one cell 12, such as the first planar image The first depth value of 30 is 2, the area value of the cell image it contains is 10, the volume of the first segment is 20, and the second depth value of the second plane image 30 is 4, which contains the cell image If the area value is 15, the second depth value needs to be subtracted from the first depth value. When calculating the volume of the section, the volume of the first section is 30, and so on, so the more depths are taken The value 22 and the plane images 30 can obtain the more accurate the volume of the at least one cell 122; in this embodiment, because the depth values 22 correspond to the plane images 30, the depths The value 22 should also correspond to the cell plane images 122 contained in the plane images 30 respectively.

In this embodiment, after scanning the detection carrier 10 and obtaining the 3D image 20, after obtaining the planar images 30 of the 3D image 20 according to the different depth values 22, the edge retouching method is used to separate the The at least one cell image 122 included in each of the two planar images 30 is calculated, and the area is calculated, and finally the planar images 30 are sequentially superimposed, using the depth values 22 and the area values of the cell images 122, Calculate the volume of the at least one cell 12 contained in the detection carrier 10. In this embodiment, the scanning range 3 can also be changed to cover a plurality of cells 12 to calculate the number of the cells 12 in the detection carrier 10 And its total volume.

In summary, the present invention provides a method for calculating cell volume, which scans a detection carrier, such as a porous microplate (or multi-well culture plate), and obtains a three-dimensional image of the detection carrier, and then distributes the cell volume according to different depth values. A three-dimensional image is obtained by obtaining a plurality of planar images of the three-dimensional image, and then edge detection is performed on the edges of the planar images, the cell images are separated, and the cell image area is calculated, and finally the planar images are sequentially superimposed, using The corresponding depth value and the area value of the cell image are used to calculate the volume of the cells in the detection carrier, and the scanning range can also be changed to calculate the number of cell colonies in the detection carrier and the total volume of the cell colonies.

Therefore, the present invention is truly novel, progressive, and available for industrial use. It should meet the patent application requirements of my country's patent law. Undoubtedly, I filed an application for a patent for invention in accordance with the law. I pray that the Bureau will grant the patent as soon as possible.

However, the above is only an embodiment of the present invention, and is not used to limit the scope of implementation of the present invention. Therefore, all the equivalent changes and modifications of the shape, structure, characteristics and spirit described in the scope of the patent application of the present invention are mentioned. All should be included in the scope of patent application of the present invention.

1: Scan lens 2: Scanning light 3: Scanning range 10: Detection carrier 12: Cell 122: Cell Imaging 20: Three-dimensional image 22: depth value 30: Flat image S2: Step S4: Step S6: Step S8: steps S10: steps

Figure 1: It is a step diagram of an embodiment of the present invention; and Figures 2A to 2D: are schematic diagrams of the steps of an embodiment of the present invention.

S2: Step

S4: Step

S6: Step

S8: steps

S10: steps

Claims (6)

A method for calculating the volume of a cell, the steps comprising: scanning a detection carrier to obtain a three-dimensional image, the detection carrier is provided with at least one cell; obtaining a plurality of plane images of the three-dimensional image according to a plurality of depth values of the three-dimensional image, the One plane image includes the at least one cell, and the plane images are parallel to each other; after performing an edge modification method on each of the plane images, at least one cell plane image included in each of the plane images is obtained ; Calculate an area of the at least one cell plane image included in the plane images to obtain a plurality of area values; and superimpose the plane images, based on the depth values and the area values, Calculate the volume of the at least one cell. The method for calculating cell volume according to claim 1, wherein the detection carrier is scanned to obtain a three-dimensional image, and the step of setting at least one cell in the detection carrier is performed by optical coherence tomography (OCT) Obtain the three-dimensional image. The method for calculating cell volume according to claim 1, wherein in the step of obtaining at least one cell plane image included in each of the plane images after performing an edge modification method on each of the plane images, the At least one cell plane image corresponds to at least one cell. The method for calculating cell volume according to claim 1, wherein in the step of calculating the volume of the at least one cell according to the depth values and the area values by superimposing the planar images, the A depth value and the corresponding area values are multiplied to calculate the volume of the at least one cell. The method for calculating cell volume according to claim 1, wherein the at least one cell line is one cell Colony or multiple cell colonies. The method for calculating cell volume according to claim 1, wherein the plurality of plane images of the three-dimensional image are obtained according to the plurality of depth values of the three-dimensional image, and the plurality of plane images include the at least one cell. A depth value is the depth value of the at least one cell in the three-dimensional image.

Images