JPH05500152A - Method and apparatus for measuring proliferation index of cell samples - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Differential dyeing is used. This system allows precise assay of cell DNA. However, its predictive power for cell replication , an important indicator of the presence of cancer (indicator L) would be improved.

It is well known that cells follow a replication cycle. Most body cells in adults are relatively sparse. It is replicating at an alarming rate and is discarded by the body and disappears through normal wear and tear. The rate is fast enough to replace cells lost through attrition. any example In most cases, these somatic cells are in the Go or resting phase of the replication cycle. that When they leave telogen, they enter G1 or first interphase, but still have extra Does not produce DNA. However, they have become more prone to stage 8, and they are filled with proliferative substances, e.g. Other substances, such as cyclin (cyc’1in) and other phase 8 proteins, arise. Cells in the synthesis or 8 stage actively synthesize DNA and during cell replication twice the amount of DNA normally contained in the cell nucleus in preparation for mitosis or division of the cell nucleus create. A normal human body cell contains 23 pairs of chromosomes and is diploid. .

Diploid is also called the 2N state. During replication, the number of chromosome pairs increases to 46; This is twice the normal amount in anticipation of cell division. The state of a chromosome just before it is replicated This is called the 4N period. The cell then enters the second interphase, or phase 62, in which DNA is rarely synthesized. Following the 02 phase is mitosis or M phase. This is where the cell itself divides. If cells continue to actively proliferate , they will enter the 61st period again. DNA analysis detects proliferating cells in normal cells or tissues. appropriate for estimating the number or proportion of cells, but this is not the case for malignant cells. It is worth noting that knowing the extent of proliferation of malignant cells is This is often necessary. This is because malignant cells are in stage 00 due to an increase in the number of chromosomes. This is because even the cells have an increased amount of DNA. Therefore, specialized cells, e.g. For example, one containing 15 times the normal NA content may be an additional chromosome or part of a chromosome. Is it a malignant cell with a cell division problem? We can confirm from DNA analysis that these are normal cells in the middle of stage 8. It is impossible to conclude from this. Therefore, regarding stage 8 proliferation and cell division process, Using other markers, such as a large variety of related proteins, DNA and It is clear that independent analytical methods have many advantages.

Quantification of cellular proliferative indices has long been used to quantify proliferative substances (proliferation) in cell samples. 5 ubstance) of cells carrying display or staining. It should also be understood that this was done by counting numbers. It is. For example, a well-known method for measuring proliferation index is Stain the cells with a binding immunofluorescent dye and manually count the stained cells to determine the proportion of cells with growth material.

Another method of measuring the proliferative index of cells is grain counting.

In that method, tritium-treated thymidine is added to the cell culture growth medium. Ru. Proliferating cells take up tritium-treated thymidine and carry it throughout the cell. Insert into the DNA being synthesized. The cells are then fixed and adjacent to the photographic emulsion. It is placed as The decay material of tritium exposes part of the base agent. exposed The parts are made visible as grains by photographic processing. Then the particles are counted It is determined whether the cells are normal or abnormal. One of the drawbacks of this method is that This means that it is very time consuming. Cells were taken alive and triturated. remain viable long enough to take up the treated thymidine. is necessary. Next, the cell is fixed adjacent to the emulsion to expose it. Retained. The relatively low intensity radiation emitted by the cells creates a latent image on the emulsion. It takes days or even weeks to obtain, after which it is developed.

One of the drawbacks of the methods in the prior art is that they cannot be used with microscope slides under high magnification. tend to introduce artifactual errors due to the monotony of counting cells on the board. This means that Those examining the slides often It is only possible to estimate the numbers associated with cells that show positive results. .

Conventional image processing systems also typically generate images of cellular objects in the processed image. however, they do not necessarily accurately determine the boundaries of the cellular object being evaluated. I am suffering from the problem of not being able to tell the difference. This means that the test is This becomes a problem when it is performed on cellular objects based on area.

Prior art methods for quantitatively analyzing cell samples for growth material are not easy to automate. will not be mobilized. Tested to control a certain number of cells with growth material This is because it is necessary to determine the baseline value for all the cells tested.

To make this type of assessment, the automated system must determine what constitutes the cell or cell nucleus. You must be able to recognize what you are doing. To solve this baseline recognition problem, To this end, the present invention employs separate staining for cell nuclei and proliferating material. Furthermore, staining are spectrally separated, so they can be fitted with optical filters. more easily distinguished. The optical separation of the two components to be measured follows the image of the cell. Makes it even more convenient to automate analysis.

Similarly, when cell object images overlap, touch, or match adjacent areas, Similar difficulties are also encountered in image analysis based on cell coverage area. In this case, what It is not possible to accurately record whether the image is actually a double or triple object image, resulting in inaccurate or inaccurate results. The argument becomes the result.

Summary of the invention The present invention provides a quick and easy method for determining the amount of growth material in a cell sample. To provide a convenient method and device. The cell sample may be a tissue sample or A cell preparation. Tissue samples are frozen sections of combined cells or paraffin This is the processed part. Cell preparations can be used to extract the brain from body fluids such as cerebrospinal fluid, blood, fluid exudates, etc. made from. Cell preparations can also be made from needle aspirates of tumor cysts, or lymph nodes. It will be done. Cell preparations can also be prepared by transferring fresh tissue pieces to microscope slides to which cells will adhere. Made from a contact preparation obtained by attaching a rotomed surface. In particular, rabbit anti Devices and methods employing mouse immunoglobulin (IgG) are Antibodies against proliferative substances such as cyclins or antigens are It is complexed with the enzyme in rust peroquinodase (HRP). Cells are HRP proliferator anti- zygotes, it has an epitope to mark them as proliferating substances. It binds only to the part of the cell that Staining, in this example, 3.3゛ Nzidine tetrahydrochloride (DAB) and hydrogen peroxide H20□ , placed in contact with cells having the antibody-HRP conjugate bound to the proliferative substance site. It's dark. HRP is a reactive compound that forms a chromogen only to the extent that it is bound. catalyze the reaction. The reaction of catalyzed chromogen formation leads to red-brown growth sites. Create a chromogen precipitate.

The cells are then counterstained, in this example ethyl green. It is commonly known as methyl green. Cell images are light The image is magnified using an optical microscope and divided into a pair of images. The divided image is a pair of narrow bands Enhanced by an optical filter. One of the narrowband optical filters is the transmission peak of the counterstain. It preferentially transmits light with wavelengths of Creates an optically enhanced growth material image with only chromogens of color. growths The background of a quality image consists of the cell nucleus and cytoplasm, which are substantially The optical density is 0. Growth material sites have a relatively high optical density. Therefore , the only easily detectable feature is the growth material site.

Other narrowband optical filters selectively transmit reddish-brown transmission peaks and counterstain peaks. chromogen, thereby emphasizing the optical density difference between the cell nucleus and the proliferating material chromogen. . The filter is optically enhanced with only background features and cell nuclei. Create an image of the cell nucleus.

The device of this invention is the first to use a monochrome television camera to Sensing material images. The enhanced cell nucleus image is captured by a second monochrome television camera. sensed by la. The analog signal representation of the image is sent to each image processing device. It will be done. Image processing equipment converts analog signals into digitized arrays of pixels and is stored in the internal frame buffer.

When a section of tissue is being examined, the device produces a growth material image with high optical density. Calculate the area of the image and perform area measurements for the growth material in that image field. Now. When a cell preparation is being tested, the device detects a threshold of proliferating material within the cells. Calculate the proliferation index on the basis of the percenteno of cell nuclei with more than the amount of Ru.

Brief description of the drawing FIG. 1 is an isometric view of an apparatus for measuring the proliferation index of a cell sample in which the present invention is carried out. , FIG. 2 is a block diagram of the device in FIG. Figure 3 is a front view of the optical conversion module of the device in Figure 1, Figure 4 is without optical filter. An enlarged view of the stained cell sample observed through the microscope in Figure 1, Figure 5 was observed through a 620 nanometer narrow band optical filter producing a cell nucleus image. An enlarged view of the sample of stained cells in Figure 4; an enlarged view of the stained cell sample of FIG. 4 observed through a data band optical filter; Figure 7 is a graph of the spectral response of the chromogen, counterstain, and narrowband optical filter; Figure 8 is a sequence of steps performed by the apparatus of Figure 1 with the cell sample analysis mode selected. Kens flowchart, Figure 9 shows the measurement of the proliferation index of a cell sample of a tissue section performed using the apparatus shown in Figure 1. Figure 10 is a flowchart of the sequence of steps performed for the cell preparation cell sample. a flowchart of the steps performed by the device in determining the proliferation index of Figure 11 is a screen representation of the tissue screen; Figure 12 is the cell preparation screen. This is the screen display.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Referring to the drawings, and in particular to FIG. It is shown. The apparatus 10 is equipped with an optical microscope 12, which is of the conventional type. However, in this example, Re1chart Diasler or Micros It is tar. Optical conversion module 14 is attached to microscope 12 and The enlarged image of the cell sample observed with the mirror 12 is optically enhanced. Figure 3 shows the most As best shown, the optical conversion module 14 is a cell nuclear image optical enhancement unit. It has a cell nucleus sensing means including a gate 16. Cell nucleus image optical enhancement unit 16 is a 620±20 nanometer red narrowband optical transmission filter 18 and a filter 18. It has a television camera 20 for receiving the filtered images.

The growth material sensing means, including the growth material optical enhancement module 22, includes a Green 500 ±20 nanometer narrow band optical transmission filter 24 and television camera 26. It also forms part of the optical conversion module 14. each television camera 20 and 26 are standard NTS with enhanced cell nucleus image and enhanced growth material image. C-compatible signal representations are respectively generated. The image processing system 28 is a television Connected to cameras 20 and 26 and enhanced cell nucleus image signal and enhanced growth material image and stores therein a cell nucleus pixel array and a growth material pixel array.

The image processing device 28 is connected to the computer 32, and in this embodiment, the image processing device 28 is IBM personal computer model A is used to process pixel arrays of nuclei and proliferating substances. Use T.

The computer 32 is provided with a system bus 34 and is connected to the image processing device 28. be done. An 80286 microprocessor 36 is connected to system bus 34.

Random access memory 38 and read only memory 40 are also used for storing information. It is connected to the system bus 34 for this purpose. The disk controller 40 is a local bus 44 to the Winchester disk drive 46 and a second information It is connected to a floppy disk drive 48 for information storage. In this example Video conversion board 50. EGA board with 256 kiloheights of memory is a command monitor 52 connected to the system bus 34 and connected to the EGA board 50; control. A keyboard processor 54 is connected to the system bus 34 and and interprets signals from a keyboard 56 connected to a code processor 54. printer 58 is connected to system bus 5-1 for communication therewith. XY or image Field board 60 is connected to system bus 34. XY board 60 In addition, a slide connected to the slide holder of the microscope 12 and related to the microscope object 64 is provided. senses the relative position of the id 62 and thus discerns the field being observed. Y position A position sensor 66 and an X position sensor 68 are also included. The Y position sensor 66 uses the communication path 70. It is then connected to the XY board 60. The X position sensor 68 connects to the XY position via the communication path 72. connected to the card 60. Microscope 12 also examines the cell sample on slide 62. an optically aligned contact with the object 74 to magnify the light forming the image; It includes an ophthalmic lens 76.

The method of the invention is carried out by collecting a cell sample, the cell sample is in the form of a tissue section consisting of frozen or paraffinized sections; Both the nucleus and cell debris, as well as the whole cell, are contained within it. Instead, Cell San A pull is caused by aspiration of blood, swollen fluid, cerebral bone marrow fluid, or the contents of a cyst or tumor. It may be a type of cell preparation that is removed. The cells in the cell sample are on slide 62. placed on top and fixed onto this. Monochrome against detected proliferative substances in cells The target antibodies are then placed in contact with them. Monoclonal antibodies are, for example, K1- 67, or 5-bromodeoxyuridine, cyclin, or cellular complex. These are antibodies against other proteins that indicate that production is occurring. monoclonal anti The body selectively attaches to all points on and within cells where proliferative substances are present.

Monoclonal antibodies can also be mixed with cross-linked antibodies and perokindase anti-peroxidase conjugates. combined into a compound. Anti-beloxidase specifically binds to the enzyme peroxidase. Contains antibodies for The peroxidase enzyme is bound to antibodies and passes through the antibody chain into cells. Retained in the growth material within.

Hydrogen peroxide and 3.3' diaminobenzidine to observe growth material sites. A quantity of a mixture containing tetrahydrochloride (DAB) is placed on the slide 62. added to the cell sample. Hydrogen peroxide and D A B react to form a reddish brown precipitate. Forms a chromogen consisting of dens. However, the normal reaction rate is relatively low. peroxida -se catalyzes the chromogen-forming reaction only at the points where perokindase is located.

Therefore, the chromogen precipitates only at points in the cell where proliferative substances are present, and the cell proliferates. Only points with growth material are selectively stained. After a period of about 15 minutes, no response DAB and hydrogen peroxide are removed from the cell sample. The cells are then methi Counterstained with Le Green (more precisely known as Ethyl Green) , it binds selectively to the cell nucleus. Therefore, the cell nucleus is stained and contains proliferative material. Dots in the cell nucleus are stained reddish-brown.

The microscope slide 62 is then placed on the holder of the microscope 12 and the object 64 is placed on it. is focused on. The light from the object 64 is passed through an eyepiece through which it can be observed by an observer. Proceed through the z. Further, the optical transducer module 14 is configured to convert the object 64 to the transducer 1 4 includes a beam splitting mirror 80 which captures about 90% of the light. That light is , to a dual prism dichroic mirror 82 that reflects a portion of the light onto the red filter 18. Added. The remaining part of the light is filtered by dichroic mirror 82 and is filtered by a green filter. data 24. Dichroic mirror 82 detects wavelengths greater than 500 nanometers. The light with wavelengths below 500 nanometers is filtered. 24. Therefore, the dichroic mirror 82 allows the light to pass through the color filter. It operates as a first color filter before reaching 18 and 24.

When light passes through filter 18, filter 18 selectively stains cells that are stained green. It blocks light from the nucleus and provides the camera 20 with a high contrast image of the cell nucleus. tortoise The module 20 then generates an NTSC cell nuclear image, which the image processor module 2 Sent to 8. The image processor module 28 has an image processor 90 and an image processor. It has a processor 92. Each of image processor 90 and image processor 92 has Da It is model AT428 from tacube Corporation. similarly , green filter 24, filter provides a high contrast growth material image to camera 26. give. Camera 26 then sends the growth material image signal to image processor 92 . Ryoga Image processors 90 and 92 digitize the analog NTSC image signal into pixels. It is equipped with an analog-to-digital converter that converts the digitized The array is then stored in an internal frame buffer. Internal frame buffer is my It is accessed via system bus 34 under the control of microprocessor 36.

The image of the cell sample observed through the eyepiece 12 has the shape shown in Figure 4. There, there is a green cell nucleus 100, a green cell nucleus 102, and a growth substance therein. A reddish-brown cell nucleus 104, a reddish-brown cell nucleus 106, and a reddish-brown and green cell nucleus 108 are depicted. It appears in the image field. As best shown in Figure 5, the cell nucleus is shown appearing through a red filter 18, and this red filter 18 is entirely It affects the green cell nucleus to become darker and more noticeable. Best shown in Figure 6 As shown, the image of the proliferating material of the cell nucleus in FIG. Shown with cell nuclei 100 and 102 bright or obscured, methyl It has a transmission peak value at approximately the same wavelength as the transmission peak value for green dyeing.

Nuclei 104, 106, and 108 with reddish-brown chromogens have clear high contrast. This is an indication of the proliferating substance that clearly appears at the end.

The cell nucleus image of FIG. 5 is stored in an internal frame buffer of image processor 90.

The growth material image of FIG. 6 is stored in an internal frame buffer of image processor 92. .

The pixel values for the image are sliced using standard image processing techniques to identify cell nuclei and It will be appreciated that the contrast between the backgrounds can be enhanced. Immediately In other words, areas of high optical density such as cell nuclei 104, 106, and 108 in Figure 6 are are shown as high optical density pixels and stored as high optical density pixels, while the background area is 110 virtually zero optical density to give a clear threshold or difference between the two areas It is stored in pixels. This system further increases the distance between the background and the nuclei being measured. This makes it easier to perform calculations and determine proliferation indices, as they can be easily distinguished. This is especially useful when This slicing technique involves an additional amplification step on the image. Work as a staff member.

Therefore, once an image is obtained by this system, it is best shown in Figure 8. , the user can determine whether the image is from a tissue section or a cell preparation. asked questions. More specifically, after the start step 120, the system 10 then executes the command The initial display screen 122 is displayed on the monitor 52, and then the step Ask the user whether the histology forms the basis for the image being processed Ru. If the knee answers in the affirmative to system 10, control will step The process moves to 126, where the tissue department screen is displayed on the command monitor 52.

If the answer is no, control transfers to step 128 where the user You will be asked if the sample came from cell preparation. If the answer is affirmative, Control passes to step 130, where the cell preparation processing and format shown in FIG. The resulting screen is displayed on the command monitor 52. No choice made If not, step 132 transfers control to a HELP screen 134. Ru.

Returning to the previous step 126, the screen of FIG. 11 is displayed during step 126. It is understood that The screen provides a menu of functions on the right side. , it is a type familiar to users of automatic cell analyzers. In particular, The user selects the Nuclear Threshold function, where the user determines whether a particular pixel value is at that point. The system 10 performs measurements for the purpose of calculating that the cell nucleus indicates the presence of a cell nucleus. The dark value optical density or pixel value can be specified using the specified value. Additionally, antibody thresholds can be similarly set. The optical density of the image in Figure 6 is measured and a threshold is set for the address of a particular pixel. is set to indicate the presence or absence of antibodies in response. Furthermore, once the threshold Once the values are set, the user then selects the nuclear-antibody mask display function. Outline and shadow of cell nucleus and antibody using c-anti masking function) Show part. Once the user has done this, the controls are further detailed in Figure 9. The process then proceeds to the tissue segment step 140 shown in FIG.

A 620 nanometer cell nuclear image in this format is received by camera 20 in step 150. be believed. The analog image signal is digitized in step 152 to determine the presence of cell nuclei. A threshold value for the present pixel is selected in step 154. Once the threshold is selected When the pixel value is lower than the darkness value, the value of the pixel is set to the preselected background level. pixels with higher values are set to high contrast for further processing. Leave the trust pixel array. The pixel array is transferred to computer system 32. , where the number of pixels with a value above the threshold of the selected nucleus is counted, Provides the amount or number of cell nuclei used as the denominator of the proliferation index in subsequent calculations .

Similarly, this type of growth material image shown in FIG. can be received by The growth material image is generated in step 162 by the image processor +92. Digitized at. selected by the user in step ]64-. The established antibody threshold reduces the background of the growth material image to zero, and the growth material antibody to effectively separate the pixel representations of. Separated pixels, i.e. of pre-selected antibodies. Pixels with values greater than the threshold are counted by the system 32 and added to the pixel counter. A count number 162 is provided in step 166.

Steps 150 to 156 calculate the area of the image field in FIG. 5 in which the cell nucleus is seen. It will be understood that effectively measuring. Steps 160 to 166 are shown in FIG. effectively measuring the area of growth material within the image field of the image field. In step 168 The computer 32 divides the growth material by the area of the cell nucleus and calculates a quotient equal to the proliferation index. arise. The proliferation index is then displayed on the tissue screen as a percentage value. Furthermore, The joint area as calculated in steps 150-156 is also displayed.

In step 128, the user , control is transferred to step 130, which is shown in FIG. Further details can be seen in step 170 as shown in FIG. At step 200 The cell nucleus image of FIG. 5 is then received by camera 20. Cell nucleus image steps Digitized at 202. Digitized cell nucleus image steps At 204, the system 10 analyzes which objects are analyzed using the adjacent labeling. determine what objects are considered to be cell nuclei and what objects are not. Ru. The object considered to be a cell nucleus is a box like the one displayed on the image monitor 30. It is shown surrounded by. In step 206, if two or more objects are in contact with each other, the operator can then He is given the opportunity to either draw boundaries or separate the images manually himself. .

In step 210, a threshold is added to the pixel array of step 208 and Image by slicing as done previously in steps 154 and 164. Amplify the difference between the elements. Similarly, in step 212, the growth material image of FIG. It can be received by Mela 26. The growth material image is digitized in step 214. , separated in step 216. The cell nucleus and proliferating material pixel arrays are arranged in step 218. The images are combined and displayed on the image monitor 30. The cell nucleus is determined by computer 32. will be counted. Similarly, the cell nucleus containing the growth material is also sent to the computer 32. will be counted more. The number of proliferating material nuclei is divided by the total number of cell nuclei and the cell preparation The sample yields a proliferation index for the sample. The proliferation index was then calculated using the cell preparation scale in Figure 12. Displayed on a clean screen.

Therefore, due to the characteristics of the tissue section in Figure 9, the proliferation index for the sample of the tissue section is clearly Stereological principles are standard in the microscopic field. It will be appreciated that it is easily and quickly calculated using . When tissue parts are not utilized and the principles of stereology are not applied, cells are Cell principal preparation technique chnique).

Furthermore, this system provides considerable amplification for the measurement of proliferation index. first amplification is when the proliferating material is identified by a chromogen and the cell nucleus is stained by a counterstain. happen. The second amplification involves passing the cell nucleus and growth material images through optical filters 18 and 24. This happens by filtering the light. Further amplification is performed on growth material and cell nuclei. Threshold high-contrast images and high-gain digital arrays for subsequent processing Occurs when configured to provide

While one particular embodiment of the invention has been shown and described, numerous changes and modifications will occur to those skilled in the art. As one may imagine, the claims herein are within the spirit and scope of the invention. It is intended to include all such changes and modifications.

Enemy wave source (Ketsumeta) [11f 7 Organization department FIG.9 Submission of translation of written amendment (Article 184-8 of the Patent Act) August 26, 1991

Claims (21)

[Claims] 1. In a device for measuring the proliferation index of a cell sample, a cell sample containing a proliferation substance is used. growth material for sensing portions of the sample and generating growth material signals in response; a sensing means for sensing a portion of the cell sample having a cell nucleus and in response to sensing a portion of the cell sample having a cell nucleus; cell nucleus sensing means for generating a nuclear signal; connected to the growth substance sensing means to receive the growth substance signal; growth material measurement for measuring the amount of said growth material from said growth material to generate a growth material amount signal; means and connected to the cell nucleus sensing means to receive the cell nucleus signal and measure the amount of the cell nucleus; a cell nucleus measuring means for generating a cell nucleus signal in response to the cell nucleus signal; and the proliferation substance. connected to a measuring means to receive the proliferating substance amount signal and connected to the cell nucleus measuring means; and receives the cell nuclear signal, and increases from the proliferative substance amount signal and the cell nuclear signal. A device comprising: a proliferation index measuring means for measuring a proliferation index. 2. The growth material sensing means limits the transmission peak of a chromogen associated with the growth material. The transmission peak of the counterstain associated with the cell nucleus transmits proliferating material and enhances the image. The apparatus for measuring a proliferation index of a cell sample according to claim 1, further comprising an optical filter. Place. 3. said cell nucleus sensing means block a transmission peak of a counterstain associated with said cell nucleus; However, the transmission peak of the chromogen associated with the proliferating material is transmitted by cell nucleus image-enhancing optics. The apparatus for measuring the proliferation index of a cell sample according to claim 1, further comprising a filter. 4. the growth material sensing means blocks a transmission peak of a chromogen associated with the growth material; The transmission peak of the counterstain associated with the cell nucleus transmits proliferating material and enhances the image. The apparatus for measuring a proliferation index of a cell sample according to claim 3, further comprising an optical filter. Place. 5. The growth material sensing means stores a digitized growth material image array. 5. The apparatus for measuring the proliferation index of a cell sample according to claim 4, further comprising means for determining the proliferation index of a cell sample. Place. 6. The growth substance measuring means measures an image area occupied by the growth substance. further comprising means for determining the amount of growth material signal occupied by the growth material. Measuring the proliferation index of the cell sample according to claim 1, which is indicative of image area. Device. 7. The cell nucleus measuring means measures an image area occupied by the cell nucleus. further comprising means for determining the image area occupied by the cell nucleus; 7. A device for measuring the proliferation index of a cell sample according to claim 6. 8. The growth material measuring means detects cells in the image field having growth material therein. further comprising means for measuring the number of nuclei, and said proliferating material amount signal indicates a cell nucleus having proliferating material; The apparatus for measuring the proliferation index of a cell sample according to claim 1, which indicates the number of . 9. The cell nucleus measuring means includes means for measuring the number of cell nuclei in the image field. further comprising: the cell nucleus signal is indicative of the cell nucleus in the image field. An apparatus for measuring the proliferation index of a cell sample according to claim 8. 10. In a method for determining the proliferative index of a cell sample, specificity for proliferative substances staining a plurality of cellular objects with a monoclonal antibody staining complex of staining the plurality of cell objects with a counterstain; and the step of staining the plurality of cell objects with a counterstain. sensing the image of the stained cells to make an enhanced image; enhancing said image of an object; Automatically determine the amount of growth material in the stained cell object from the enhanced image a method comprising the steps of: 11. 11. The proliferative substance comprises an antigen such that Ki-67 is an antibody. A method for measuring the proliferation index of cell samples. 12. 11. The cell according to claim 10, wherein the growth material comprises 5-bromodeoxyuridine. A method for measuring the proliferation index of cell samples. 13. The proliferation index of a cell sample according to claim 10, wherein the proliferation substance comprises a protein. How to measure. 14. Proliferation of a cell sample according to claim 10, wherein the proliferation substance comprises cytalin. How to measure index. 15. 11. The monoclonal antibody staining complex comprises peroxidase. A method for measuring the proliferation index of cell samples. 16. 15. The monoclonal antibody staining complex further comprises an anti-peroxidase. A method for measuring the proliferation index of a cell sample as described in . 17. Claim 1, wherein the monoclonal antibody staining complex further comprises diaminobenzidine. 6. A method for measuring the proliferation index of a cell sample according to 6. 18. In a system that performs image enhancement operations on images of multiple cell objects, multiple a sensor for sensing an image of a cellular object and generating an image signal in response thereto; means and a mask control means for receiving a mask control signal from a user; and the mask control the signal has a first state and a second state; and the cell object image is preselected. and the mask control signal is in the second state. a mask generation means for automatically generating a mask for each cell object image; , within the designated image field when the mask control signal is in the first state. Cell object tags to generate user-perceivable indications for each cell object. and The means for receiving the mask signal is configured to detect the details identified by the cell object tag means. The mask generation function means for withholding, and a system comprising: 19. Analyzing predetermined characteristics of multiple cells in a tissue section or cell preparation In the system for Test multiple cells in a cell sample and display multiple cells in the image field means for generating an image signal representation of a surface image; and means for storing said image signal. storage means connected to the optical means; and a storage means connected to the optical means; A mode for determining whether or not to be evaluated based on the cell image generated from the cell image. a selection means, a predetermined characteristic connected to the storage means and the mode selection means; a cell analysis means for quantitatively analyzing the cell image for sex, and the predetermined cell image. output means for outputting the results of the quantitative analysis of the determined characteristic. 20. In a system for assaying the amount of growth material in a cell sample, Test multiple cells in a sample and display multiple cell panes in the image field optical means for generating an image signal representation of an image and for storing said image signal; storage means connected to the optical means of the tissue section or cell preparation; asking the user whether he or she would like to be evaluated on the basis of said cell image arising from Assay mode selection that generates a tissue site selection signal or cell preparation selection signal in response. means connected to the storage means and the test mode selection means to select the test mode based on the area; Measuring the amount of proliferative material in the cell sample and generating the assay; said tissue selection means; is the tissue selection means that is activated in response to the selected tissue selection and the verification mode. a positive response to the growth substance; The above details are based on the recording of a large number of stored cell images to generate the assay. cell preparation means for determining the amount of said growth material in a cell sample; and said assay. and an output means for outputting the results to a user. 21. In a system that performs image enhancement operations on images of cell samples, multiple comprising sensor means for detecting an image of the cell and generating an image signal responsive thereto; an image analysis and decomposition means, Define a shared cell image resulting from images of a pair of cells that share adjacent parts. means to To separate the divided cell images into a pair of separate cell images for subsequent processing. a means for determining the boundaries of the divided cells; and a means for displaying the boundaries of the divided cells. A system comprising means for: