CN111768424A - Cell image extraction method applied to linear array detector flow cytometer - Google Patents
Cell image extraction method applied to linear array detector flow cytometer Download PDFInfo
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Abstract
The invention discloses a cell image extraction method applied to a linear array detector flow cytometer, which comprises the following steps of 1, establishing an initial theoretical background model of a cell image; step 2, updating the initial theoretical background model to obtain an improved background model; step 3, separating the foreground cells from the background by improving the background model to obtain the change information of the foreground cells; and 4, extracting the linear array scanning image of the foreground cell, and finishing the extraction of the cell image. The invention can simplify and even eliminate the problems of poor imaging effect, complex background and the like caused by the influence of the quality of the light source and the micro-channel in the experimental operation process; the difficulty of later cell analysis and treatment is effectively reduced.
Description
Technical Field
The invention belongs to the technical field of cell detection, and relates to a cell image extraction method applied to a linear array detector flow cytometer.
Background
Cell detection plays an important role in medical research and in many disease monitoring. However, the conventional cell detection instrument is large in size and expensive, and is only suitable for large medical institutions. The micro-fluidic technology is combined with the image processing technology, so that the miniaturization, low cost, easy operation and popularization of the cell detection system become possible. The cell image acquisition device based on the micro-fluidic technology and the linear array detector can solve the traditional problems, but the imaging effect of the device is easily influenced by the quality of a light source and a micro-channel, so that the cell analysis and processing difficulty in the later period is high, and therefore, the cell image acquisition of the linear array acquisition system is an important part in the cell analysis and processing in the later period.
Disclosure of Invention
The invention aims to provide a cell image extraction method applied to a linear array detector flow cytometer, which solves the problem of poor cell image imaging recovery effect caused by complex background of acquired cell images.
The invention adopts the technical scheme that a cell image extraction method applied to a linear array detector flow cytometer is implemented according to the following steps:
step 3, separating the foreground cells from the background by improving the background model to obtain the change information of the foreground cells;
and 4, linearly mapping the change information of the foreground cells to an acquired image of the linear array scanning imaging system, extracting the linear array scanning image of the foreground cells, and finishing the extraction of the cell image.
The present invention is also characterized in that,
the cell image is obtained by acquiring image data of cells flowing through a detection area of the linear array detector at a constant speed at a fixed frame frequency and splicing each frame of data of the image data according to a time sequence.
Initial theoretical background model P'iP with respect to pixel valueiThe function expression is P'i＝g(Pi) Initial theoretical background model P'iExtracted information on changes in foreground cells C1Pi＝Pi-P’i。
The step 2 is implemented according to the following steps:
step 2.1, according to the foreground cell change information C1PiSetting a single or a plurality of threshold values for the cell image with the cell flowing through for threshold segmentation, realizing the rough segmentation of the foreground cell image of the cell image with the cell flowing through, obtaining the Mask of the background image, and recording as: MaPi；
Step 2.2, use mask MaPiExtraction of item FiFiltering out the pixels with cells from the background image of the row, and determining the F-th image of the cell image with cells flowing throughi-NTo FiLine mean background model P "iMean background model P "iI.e. to improve the background model, P "iIs a function expression P'i＝h(P’I,MaPi)。
Step 3 specifically comprises the step of providing an initial theoretical background model P'iReplacement by an improved background model P "iFor the improved background model P "iThe extracted information of the foreground cell change is C2Pi＝Pi-P”i。
The invention has the beneficial effects that:
the invention relates to a cell image extraction method applied to a linear array detector flow cytometer, which comprises the steps of carrying out foreground threshold segmentation on the basis of a mean background model, updating the model in real time, mapping the change information of foreground cells to an image for foreground extraction, and obtaining a linear array scanning image of the foreground cells; therefore, the problems of poor imaging effect, complex background and the like caused by the influence of the quality of the light source and the micro-channel in the experimental operation process can be simplified or even eliminated; the method effectively reduces the difficulty of later cell analysis and processing, and is an extremely important step for later cell image analysis and processing.
Drawings
FIG. 1 is a schematic diagram of a flow cytometer based on a linear array detector;
FIG. 2 is a side view of the basic structure of a flow cytometer based on a line detector;
FIG. 3 is a cell image of a cell image extraction method of the present invention applied to a linear array detector flow cytometer;
fig. 4 is a diagram of change information of foreground cells obtained after separating a foreground and a background in a cell image extraction method applied to a linear array detector flow cytometer of the present invention;
fig. 5 is a foreground cell image obtained by separating a foreground and a background based on one-dimensional viewing in a cell image extraction method applied to a linear array detector flow cytometer according to the present invention;
fig. 6 is a linear array scanning image of foreground cells in a cell image extraction method applied to a linear array detector flow cytometer of the present invention.
In the figure, 1, a microfluidic chip, 2, a linear array detector, 3, a data processing device, 4, a parallel light source, 5, a microchannel, 6, a microfluidic channel inlet a, 7, a microfluidic channel inlet b, 8 and a microfluidic channel outlet.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention relates to a cell image extraction method applied to a linear array detector flow cytometer, a flow cytometer based on a linear array detector is shown in figure 1-2 and comprises a microfluidic chip 1, a linear array detector 2, a data processing device 3, a parallel light source 4, a microchannel 5, a microfluidic channel inlet a6, a microfluidic channel inlet b7 and a microfluidic channel outlet 8, and the cell image extraction method is implemented according to the following steps:
Mean value modeling is carried out on cell images when no cell flows through, and F is setiFor reading ith frame of linear array scanning image, let PiIs the FiLine by line pixel values, P'iIs the Fi-NGo to FiMean model of lines, N<i and both N and i are natural numbers other than 0, and buffering the image data F according to the frame streami-NTo FiAnd frames, wherein N is the current frame, adding pixel points of the N frames and averaging the pixel points to obtain an initial theoretical background model P'i；
Initial theoretical background model P'iP with respect to pixel valueiThe function expression is P'i＝g(Pi) Initial theoretical background model P'iExtracted information on changes in foreground cells C1Pi＝Pi-P’i。
The cell image is obtained by acquiring image data of cells flowing through a detection area at a constant speed at a fixed frame frequency through a linear array detector and splicing each frame of data of the image data according to a time sequence as shown in fig. 3;
step 2.1, according to the foreground cell change information C1PiSetting a single or a plurality of threshold values for the cell image with the cell flowing through for threshold segmentation, realizing the rough segmentation of the foreground cell image of the cell image with the cell flowing through, obtaining the Mask of the background image, and recording as:MaPi；
step 2.2, use mask MaPiExtraction of item FiFiltering out the pixels with cells from the background image of the row, and determining the F-th image of the cell image with cells flowing throughi-NTo FiLine mean background model P "iMean background model P "iI.e. to improve the background model, P "iIs a function expression P'i＝h(P’I,MaPi)。
Step 3, separating the foreground cells from the background by improving the background model to obtain the change information of the foreground cells;
an initial theoretical background model P'iReplacement by an improved background model P "iFor the improved background model P "iThe extracted information of the foreground cell change is C2Pi＝Pi-P”iThe foreground cell change information map is shown in FIGS. 4-5.
Will improve the background model P "iInformation on the change of the foreground cells C2PiMapping to cell image collected by linear array detector scanning imaging system according to C2PiAnd compressing the background dynamic range and stretching the cell dynamic range to obtain the linear array scanning image of the foreground cells, so as to finish the extraction of the cell image.
For example, the linear array scan image of the cell image shown in fig. 3 and the linear array scan image of the foreground cell shown in fig. 6 obtained by the cell image extraction method based on the linear array detector flow cytometer of the present invention can be clearly seen, the foreground cell information is more clear, and the problems of complex background and the like caused by the influence of the light source and the quality of the microchannel during the experimental operation process are simplified and even removed, thereby reducing the difficulty of the later cell analysis and processing.
Claims (7)
1. A cell image extraction method applied to a linear array detector flow cytometer is characterized by comprising the following steps:
step 1, establishing an initial theoretical background model of a cell image;
step 2, carrying out rough segmentation on foreground cells and a background of the cell image, extracting a background image, and updating an initial theoretical background model to obtain an improved background model;
step 3, separating the foreground cells from the background by improving the background model to obtain the change information of the foreground cells;
and 4, linearly mapping the change information of the foreground cells to an acquired image of the linear array scanning imaging system, extracting the linear array scanning image of the foreground cells, and finishing the extraction of the cell image.
2. The cell image extraction method applied to the linear array detector flow cytometer as recited in claim 1, wherein the cell image is obtained by collecting image data of cells flowing through a detection area of the linear array detector at a constant speed at a fixed frame frequency and splicing each frame of data of the image data according to a time sequence.
3. The method as claimed in claim 1, wherein the step 1 is specifically that mean value modeling is performed on cell images when no cells flow, and F is setiFor reading ith frame of linear array scanning image, let PiIs the FiLine by line pixel values, P'iIs the Fi-NGo to FiMean model of lines, N<i and both N and i are natural numbers other than 0, and buffering the image data F according to the frame streami-NTo FiAnd frames, wherein N is the current frame, adding pixel points of the N frames and averaging the pixel points to obtain an initial theoretical background model P'i。
4. The cell image extraction method applied to the linear array detector flow cytometer as recited in claim 3, wherein the cell image extraction method is characterized in thatOf the initial theoretical background model P'iP with respect to pixel valueiThe function expression is P'i＝g(Pi) Initial theoretical background model P'iExtracted information on changes in foreground cells C1Pi＝Pi-P’i。
5. The cell image extraction method applied to the linear array detector flow cytometer as claimed in claim 4, wherein the step 2 is specifically implemented according to the following steps:
step 2.1, according to the foreground cell change information C1PiSetting a single or a plurality of threshold values for the cell image with the cell flowing through for threshold segmentation, realizing the rough segmentation of the foreground cell image of the cell image with the cell flowing through, obtaining the Mask of the background image, and recording as: MaPi；
Step 2.2, use mask MaPiExtraction of item FiFiltering out the pixels with cells from the background image of the row, and determining the F-th image of the cell image with cells flowing throughi-NTo FiLine mean background model P "iMean background model P "iI.e. to improve the background model, P "iIs a function expression P'i＝h(P’I,MaPi)。
6. The method for extracting cell images applied to a linear array detector flow cytometer as claimed in claim 5, wherein the step 3 is to use an initial theoretical background model P'iReplacement by an improved background model P "iFor the improved background model P "iThe extracted information of the foreground cell change is C2Pi＝Pi-P”i。
7. The method for extracting cell images applied to a linear array detector flow cytometer as claimed in claim 6, wherein the step 4 is to improve a background model P "iInformation on the change of the foreground cells C2PiMapping to cell image collected by linear array detector scanning imaging system according to C2PiAnd compressing the background dynamic range and stretching the cell dynamic range to obtain the linear array scanning image of the foreground cells, so as to finish the extraction of the cell image.
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