WO2001046671A1

WO2001046671A1 - Apparatus and method for image analysis quality control and assurance

Info

Publication number: WO2001046671A1
Application number: PCT/CA2000/001563
Authority: WO
Inventors: Paul A. Presta; Daniel S. Maclean
Original assignee: Veracel Inc.
Priority date: 1999-12-20
Filing date: 2000-12-20
Publication date: 2001-06-28
Also published as: AU2336501A

Abstract

A test slide apparatus for assessing stain quality of a biological specimen in an automated analysis system. The test slide comprises a slide carrier, a micro-porous layer applied to the surface of the slide carrier, a stain-reactive element deposited in the micro-porous surface. The stain-reactive elements may comprise one or more encapsulated proteins and include synthetic DNA, synthetic RNA, keratin, or albumin, or a naturally encapsulated protein cultured from an immortal cell line, such as MCF-7 cells.

Description

TITLE: APPARATUS AND METHOD FOR IMAGE ANALYSIS QUALITY CONTROL AND ASSURANCE

FIELD OF THE INVENTION

The present invention relates to medical and biological specimen analysis, and more particularly to an apparatus and method for image analysis quality control and assurance.

BACKGROUND OF THE INVENTION

The collection, preservation, mounting, and analysis of tissue of cellular specimens is an important part of medical and biological diagnostic systems. When collected as exfoliated cells, this process falls under the category of cytology. If the specimen is collected and processed as intact tissue, for example in a biopsy, then the process is within the field of histology. Cells gathered from the bloodstream generally form part of a hematological analysis.

The Pap test is a well known procedure under the category of cytological specimen and analysis. The Pap test is based on the Papanicolaou staining protocol. The first step in a Pap test involves scraping the cells from the uterine ectocervix for precancerous lesions of the uterine cervix, and comprises the specimen collection phase. Next, the Pap test cells gathered from the ectocervix are 'smeared' onto a microscope slide and then fixed. The fixation procedure kills and preserves the cells in such a way as to render them inert and representative of their appearance in life and results in a Pap smear for the specimen. The Pap smear is then sent to a laboratory where the preparation and analysis of the specimen is completed. The preparation steps in the laboratory include the staining of the cells and covering the stained cells with a cover slip for protection. According to the Papanicolaou protocol, the nuclei are stained dark blue while the cytoplasm is stained anything from a blue-green to an orange-pink. The Papanicolaou stain is a combination of several stains or dyes together with a specific protocol designed to emphasize and delineate cellular structures of importance for pathological analysis. The stains or dyes included in the Papanicolaou Stε in are Haematoxylin, Orange G and Eosin Azure (a mixture of two acid dyes, Eosin Y and Light Green SF Yellowish, together with Bismark Brown). Each stain component is sensitive to or binds selectively to a particular cell structure or material. Haematoxylin binds to the nuclear material colouring it dark blue. Orange G is an indicator of keratin protein content. Eosin Y stains nucleoli, red blood cells and mature squamous epithelial cells. Light Green SF yellowish acid stains metabolically active epithelial cells. Bismark Brown stains vegetable material and cellulose. The purpose of the staining procedure is to render the cells visible under magnification and to allow differentiation of various intra-cellular components or compounds.

The typical Pap smear prepared according to this procedure comprises a complex arrangement of cells, debris, blood, leukocytes, bacteria, fungus, and other components, as depicted in Fig. 1 (a). Under the skilled eye of an experienced cytologist, conventional Pap smears are highly effective for screening for cervical cancer and other malignant cell forms. While there are great advantages and efficiencies to be achieved through the utilization of automated image analysis, the complex arrangement of cells in a conventional Pap smear complicate the application of automated image analysis.

One approach followed in the development of automated cytological image analysis systems is the utilization of controlled specimen preparations such as a monolayer specimen and a liquid-based preparation (LBP) as shown in Fig. 1 (b). The advantage of the monolayer specimen preparation technique is the ability to exert control over the physical arrangement of the cells on the microscope slide. By effectively narrowing the tolerances on the physical arrangement of the cellular components, the accuracy and reproducibility of the automated analysis systems is greatly enhanced. One problem that remains with the application of automated analysis systems is the variations which arise as a result of the staining of these cells. Cytological staining remains more of an art than a science. There are many reasons for this. In some cases it is because the stains themselves are poorly manufactured. In other cases it is the technique that is to blame since there is a wide range of parameters to control and few ways of ensuring that all are within acceptable levels.

Automated cytological image analysis systems rely on the precise measurement of various features associated with intra-cellular components, such as the nucleus, and these may easily be skewed by poor stain control. For example, overstaining the nucleus of a cell alters the appearance of its chromatin texture, which is one the most important analytical features. Similarly, understaining can compromise the segmentation of the nucleus and thereby affect morphological measures such as its area and perimeter. Thus, the measurement and control of stain variations in cytological, and also histological and hematological, specimens is a crucial issue in the application of automated image analysis for medicine and biology.

Accordingly, there remains the problem of measuring and controlling stain variations in cytological, histological, hematological, and other types of specimens, particularly in the field of automated image analysis for medicine and biology.

BRIEF SUMMARY OF THE INVENTION

The present invention provides an apparatus and method for image analysis quality control and assurance suitable for use with automated cytology analysis systems such as the those from the common assignee of the subject invention. According to one aspect, the present invention provides a previously stained specimen comprising an encapsulated protein which is deemed to be an optimally stained specimen. The encapsulated protein is placed on a microscope slide, stained and its appearance, and other characteristics, are characterized using the automated analysis system. The slides previously deemed 'optimally stained' provide a benchmark for the analysis of the stain quality in terms of segmentation results or features extracted by the automated analysis system.

In a first aspect, the present invention provides an apparatus for assessing stain quality of a biological specimen in an automated analysis system, the apparatus comprises: (a) a slide carrier; (b) a layer applied to a surface of the slide carrier, the layer forming a micro-porous surface; (c) a stain- reactive element, the stain-reactive element being deposited in at least some of the micro-pores in the layer on the surface of the slide carrier.

In a second aspect, the present invention provides a method for verifying the operation of an automated analysis system, the method comprises the steps of: (a) providing a test slide for the automated analysis system, the test slide comprising: (i) a slide carrier; (ii) a layer applied to a surface of the slide carrier, the layer forming a micro-porous surface; (iii) a stain-reactive element, the stain- reactive element being deposited in at least some of the micro-pores in the layer on the surface of the slide carrier; (b) applying the test slide to the automated analysis system and operating the automated analysis system to perform selected processing procedures on the test slide to produce one or more test results for the test slide; (c) recording the test results for the test slide so that the stain-reactive element provides a predictable response for the operation of the automated analysis system.

Other aspects and features of the present invention will become apparent to those ordinarily skilled in the art upon review of the following description of specific embodiments of the invention in conjunction with the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

Reference will now be made to the accompanying drawings, which show, by way of example, a preferred embodiment of the present invention, and in which:

Fig. 1 (a) shows a conventional smear of the type prepared in gynaecologic cytology;

Fig. 1 (b) shows a conventional liquid-based smear of the type prepared gynaecologic cytology;

Fig. 2 shows typical cell type variations present in the conventional liquid- based smear of Fig. 1 (b);

Figs. 3(a) to 3(d) shows an apparatus and processing steps for synthetic encapsulation of a protein according to one embodiment of the present invention;

Fig. 4 shows a test slide with encapsulated proteins and staining according to the present invention;

Figs. 5(a) to 5(b) shows an apparatus and processing steps for synthetic encapsulation of a protein according to another embodiment of the present invention; and

Fig. 6 shows a natural encapsulation protein mechanism according to another embodiment of the invention; and Fig. 7 shows in di grammatic form an automated image analysis system of the type suitable for use with the apparatus and method of the present invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Reference is made to Fig. 2, which shows cell type variations in the liquid- based preparation for a typical Pap smear (i.e. prepared according to the Papanicolaou Staining protocol) specimen denoted generally by reference 10, i.e. the gynaecologic cytology specimen of Fig. 1 (b). The cell type variations in the Pap smear include squamous intermediate type cells denoted generally by reference 11 , polymorphonuclear leukocyte type cells denoted generally by reference 12, squamous metaplastic type cells denoted generally by reference 13, and debris denoted generally by reference 14.

The specimen is stained according to the well-known Papanicolaou (i.e. Pap) staining protocol. The cells in the specimen 10 that respond to the stain comprise naturally-derived, encapsulated protein and this is what the stains will generally bind to. For example, in the Pap stain, the Haematoxylin will bind to DNA or RNA in the cell nucleus, and the Orange G stain will selectively bind to keratin in the cytoplasm, etc.

The problem with using a Pap smear specimen on its own, such as depicted in Fig. 2, is that there are many cell types, for example 11-14, present in a typical exfoliated specimen.

As will now be described, the present invention provides a previously deemed optimally stained specimen comprising an encapsulated protein. The encapsulated protein is placed on a microscope slide, stained and its appearance, and other characteristics, are characterized using an automated system, for example an automated image analysis system as described below with reference to Fig. 7. The encapsulated protein comprises a synthetic encapsulated protein or a naturally encapsulated protein that has a predictable character as will also be described in more detail below.

Reference is made to Fig. 3(a), a synthetic encapsulation protein test slide 100 according to one aspect of the present invention. As shown, the test slide 100 comprises a slide 101 , for example a glass slide, with a micro-porous film or layer 102 applied or formed on a surface of the slide 101. The micro- porous film 102 receives proteins indicated by reference 104, which are deposited as described below.

The micro-porous film or layer 104 may be formed in a number of ways. For example, the glass for the slide 102 can be made using the sol-gel process which does not involve melting. The sol-gel process can be used to create an inorganic silicate glass slide 101 with a porous matrix which forms the micro- porous film 104. In the sol-gel process, organic compounds such as alcoholates of silicon, sodium or calcium are used. With water, these compounds split into water and alcohol in a process of hydrolysis which also creates a structure where the metallic atoms are bonded to oxygen atoms in an irregular non-crystalline network, thus forming a gel. Low-temperature treatment turns this gel into an inorganic glass slide with a porous matrix of the form shown in Fig. 3(a). The proteins to be encapsulated can then be inserted into the micro-porous layer 102.

The next step as shown in Fig. 3(b) involves applying or encapsulating proteins 104 to the micro-porous film 102 on the slide 101. The proteins 104 for encapsulation can be 'painted' or 'printed' on the film 102 using ink-jet technology, for example. As shown in Fig. 3(b), the proteins 104 are emitted from an ink-jet nozzle 110 onto the micro-porous surface 102 and the proteins 104 settle into micro-pores in the film 102. The proteins 104 are deposited in groups 106, indicated individually as 106a, 106b and 106c, and preferably arranged in a regular pattern as shown in Fig. 3(c). The encapsulation process may be completed with subsequent processing of the sol-gel glass or by over- coating the micro-porous film 102 with another porous compound as shown in Fig. 3(d).

Several different proteins may used for the slide 100. For example, suitable proteins include synthetic DNA or RNA (stretches of the double helix manufactured by commonly available techniques), keratin, albumin, etc. In one application for the test slide according to this aspect of the invention, these different types of proteins 104 are encapsulated on a slide 118 as described above and arranged in specific regions 120 as shown in Fig 4. The test slide 118 is then processed by the laboratory in the usual manner, for example, according to the Papanicolaou (Pap) staining protocol. The slide 118 is applied as a 'test specimen' to an automated analysis system 200 as shown in Fig. 7 and described in commonly assigned International Patent Application filed underthe title "Controlled Review of Medical Sample" on October 27, 2000, the disclosure of which is hereby incorporated by reference. Referring to Fig. 4, the test slide 118 preferably includes a label 122 with a machine readable identifier 124 such as a barcode. The machine readable identifier 124 facilitates the automatic identification of the test slide 118 when it is applied to the automated image analysis system 200. The automated analysis system 200 examines the different regions 120 of the encapsulated proteins and assesses each region 120 of protein against the results stored from previous 'optimally' stained specimens. The slide 1 18 may also be used as a standard resolution test target.

Reference is next made to Fig. 5(a) which shows a variation of a test slide 130 according to the present invention. For the test slide 130, the proteins 104 are deposited in the micro-porous film 132 and arranged in groups 134, shown individually as 134a, 134b, 134c, 134d, 134e, in varying densities. The proteins 104 may be deposited in the varying amounts 134 as a part of a local stain scale.

Another variation of the test slide 140 is shown in Fig. 5(b). The test slide 140 comprises a slide 141 and a micro-porous layer or film 142. Proteins 144 are encapsulated in the micro-porous layer 142 according to the process as described above. As also described above, an overcoat layer 146 is applied, and according to this aspect of the invention, the overcoat layer 146 is applied in various thicknesses indicated individually as 146a, 146b, 146c, and 146d, in Fig. 5(b). After the encapsulated proteins 144 in the test slide 140 are exposed to the stain protocol, the test slide 140 may used in an automated analysis system (Fig. 7) to not only evaluate the suitability of the stain processing, but also to identify the stain components that are somehow compromised. The test slide 140 may also be used as a standard resolution test target and to supply dynamic range information.

In accordance with another embodiment of the invention, naturally encapsulated proteins are utilized in a test slide instead of the synthetic encapsulation process as described above. The naturally encapsulated proteins may comprise cells that are cloned from a single cell to produce a monotype culture of cells, or more simply, the naturally encapsulated proteins may comprise a monotype derived from an immortal cell line such as a cancer. A suitable immortal cell line is MCF-7which derived from a breast cancer as depicted in Fig. 6. All of the cells have identical appearance and structure when harvested correctly using standard, widely understood procedures. In practical terms, the immortal cell line would be cultured and harvested to produce a suspension of the cells in an appropriate preservative solution. If this cell type on its own is not sufficient to provide an evaluation of all of the relevant parameters, a second or third cell type may be cultured and added to the mix. This suspension would be used to make a monolayer specimen on a test slide utilizing known LBP procedures. After being appropriately labelled, the monolayer slide specimen provides the stain 'test' slide.

The test slide accompanies the other specimens through the staining procedure used in the laboratory. After insertion into the automated analysis system 200 (Fig. 7), the unique label identifies the slide as a 'test' specimen and the automated analysis system 200 treats it accordingly. For example, the automated analysis syst m 200 could perform image segmentation and feature extraction on the test slide and then compare these results to those of an optimally-stained slide held somewhere in its memory. The extent and type of deviation from the optimal characteristics allow the automated analysis system 200 to reject slide from a batch that are improperly stained or to alert the laboratory to incipient problems with the stain technique before they become detrimental to the automated analysis.

Reference is next made to Fig. 7 which shows the automated system 200 in more detail. The automated system 200 comprises an automated cell classifier for performing the analysis. The automated system 200 includes a turntable 202 for manipulating slides, i.e. including the test slide 100 with the encapsulated protein, a slide dotter 204 for placing marks on the slide, hardware including a processor and memory storage device 206, a camera 208, and a light source 210. A robotic arm (not shown) places the slide in a predetermined position on the scanning turntable 202, which is motorized for manipulating the position of the slide. The barcode, for example 124 on the test slide 118 (Fig. 5(a)), is read by an optical scanner (not shown) in the automated system 200 and a file corresponding to the slide identification number on the barcode is created in the memory storage device 206. The slide dotter 204 places a physical reference mark on the slide at a predetermined reference position. The automated cell classifier maps out fields of view on the slide and the location of each field of view with respect to the reference position.

In certain aspects of the automated system 200, the light from the illuminated specimen is separated into different predetermined spectral bands by a prism assembly associated with the light source. The camera 208 comprises three cameras and each camera captures images in one of the predetermined spectral bands. A controller controls the light source and cameras and a processor converts the images into data suitable for processing. The system for capturing images of the slide may comprise a system according to certain embodiments disclosed in commonly assigned International Patent Application Nos. WO 98/52016 and WO 98/35262.

Next, the images are analyzed by the automated cell classifier of the apparatus. The automated cell classifier performs an analytical scan of the slide and identifies cells which may be irregular. For example, preferred automated cell classifiers are described in commonly assigned International Patent Application Nos. WO 98/30317; WO 98/22909; WO 97/11350; WO 97/04419; WO 97/04418; WO 97/04348; and WO 97/50003, the disclosures of which are hereby incorporated by reference. Preferred devices are also disclosed in commonly assigned United States Patent Nos. 5,532,874 and 5,708,830, the disclosures of which are hereby incorporated by reference. Other preferred devices are disclosed in commonly assigned U.S. Patent Application of Ryan S. Raz and louri Lappa, entitled "Database Element Retrieval By Example", Serial No. 09/115,608, filed July 15, 1998, the disclosure of which is hereby incorporated by reference.

The automated cell classifier 200 digitizes the images captured by the camera 208 and the images are manipulated using known techniques. Preferably, the techniques used are techniques disclosed in certain embodiments of the publications incorporated by reference above, such as, for example, International Patent Application Nos. WO 98/22909 and WO 97/1 1350, the disclosures of which are hereby incorporated by reference. The images captured by the camera 208 comprise data concerning cells and background material in each of the fields of view. Furthermore, each cell comprises nuclear and cytoplasmic material. The data is analyzed to determine which portions of the images represent background material, cytoplasmic material, or nuclear material. Segmentation is performed by the automated cell classifier 200 to distinguish the background, nucleus and cytoplasm within each image. Preferred methods of performing segmentation are disclosed in certain embodiments of International Patent Application Nos. WO 98/22909, WO 97/1 1350 and WO 97/04418, the disclosures of which are incorporated by reference. The automated cell classifier 200 performs feature extraction, in which each nucleus identified during segmentation is analyzed separately. The automated cell classifier 200 uses neural networks and algorithms to reach a classification forthe nuclei identified during segmentation. The classification may be performed utilizing a neural network or elliptical basis function, as known by those of ordinary skill in the art. The classifications preferably comprise a score, such as, for example, a score on a scale of 0 to 1 , where 0 identifies nuclei of benign cells and 1 identifies nuclei of abnormal cells. Thus, in preferred embodiments of the automated system 200, the classifications developed by the automated cell classifier comprise a score representing the likelihood that the nucleus is abnormal or benign.

The test slides according to the present invention as described above find particular application for (1 ) specimen preparation quality control/assurance and for (2) automated analysis system integrity assurance.

(1 ) Specimen Preparation Quality Control/Assurance

In this application for an automated image analysis system, the preparation of a monolayer specimen, including its staining characteristics, is evaluated, verified and quantified using a test slide with a naturally encapsulated protein set, for example, the breast cancer cell line, MCF-7, as described above.

For the test slide, the immortal cell line derived from MCF-7 breast cancer cells can be commercially obtained from the American Type Culture Collection (ATCC). These cells are raised and harvested using standardized techniques in a typical biological laboratory. The harvesting of the cells, for example using the enzyme trypsin, produces many hundreds of thousands of identical cancer cells. The harvested cells are placed in suspension using an appropriate preservative solution and vial that is compatible with the LBP technique used by the laboratory. The suspension of a monotype of cancer cells is then used by the laboratory to create one or more test slides (as described above) according to the present invention in the form of monolayer specimens. The test slides are labelled and included in the batch staining procedure used for the actual medical specimens on a regular basis. The use of these test slides on a regular basis allows the laboratory to create a time-series so that the quality of the specimen preparation procedure may be tracked over time.

After recognizing its special label, each the test slides is analyzed by the automated image analysis system according to predefined quality assurance procedures. The first quality assurance procedure may comprise quantifying the density of the cells on the test slide. The quantification step can be done quickly by any image analysis system and comprises using a gross calculation of the area covered by the cells. By dividing this area by the known area of a single cancer cell, the total number of cells on the test slide can be determined. This information provides an important indicator of the quality of the specimen deposition technique and can alert the laboratory to problems with that technique that might jeopardize the diagnostic value of the preparation.

The second quality assurance procedure applied to the test slide by the automated analysis system comprises estimating the success of the cell segmentation, i.e., the accurate separation of the cell nucleus from its cytoplasm. The monotype (or multiple monotypes) nature of the specimen on the test slide facilitates this analysis. This is because the area of the cell occupied by the nucleus, and the perimeter of the nucleus, are known in advance to great precision. After the segmentation is complete, the total area occupied by the nucleus, for any one cell type, may be compared to the expected results. This may, for example, indicate whether or not the cells are over or understained. Further analysis of the variations in cell nucleus area, perimeter, shape, etc. might be properly correlated to specific staining shortcomings and so provide the laboratory with the information it requires to correct a problem. Furthermore, the type and extent of the stain variations may allow the automated analysis system to 'adjust' its analysis accordingly so as to accommodate these variations found in the 'true' medical specimens providing that the variations are not so extreme as to render the results completely inaccurate.

Finally, after the segmentation operation is completed, the full range of extracted features may be used to further analyze the specimen on the test slide and to decide whether the stain variations are within the allowable band of variations or if they cannot be accommodated by the automated image analysis system.

(2) Automated Analysis System Integrity Assurance

In this application, the test slide 100 with the synthetic encapsulated protein set as described above with reference to Figs. 3 and 5 is utilized. Advantageously, the test slide 100 can serve the dual purpose of both verifying the stain quality and verifying the image resolution and positioning accuracy for the automated analysis system.

For this application, the objective is to create the test slide 100 with a set of encapsulated proteins whose density and overcoating can provide a scale for stain uptake. The proteins are physically arranged on the microscope slide to create a set of optical resolution targets to determine the quality of the optics of the automated image analysis system. Alternatively, the encapsulated proteins may co-exist with an optical target created by standard technique (i.e. metal- evaporation, etc.)

The test slide 100 with the encapsulated protein set is then handled in precisely the same manner as a standard medical or biological specimen, i.e. the test slide 100 and encapsulated protein set is stained and cover-slipped in the usual fashion and submitted to the automated analysis system for review. The test slide 100 is submitted to the automated analysis system and the bar code encounters the test specimen, the machine readable bar code 124 (Fig. 4) on the slide 100 alerts the automated image analysis system to the purpose of the test slide 100. The automated analysis system then executes a pre-determined range of analytical procedures to gauge the stain uptake in each of the different protein groups 120 (Fig. 4) used on the test slide 100. In this application, it is not necessary for the automated analysis system to subject the specimen to the 'segmentation' or 'feature extraction' operations. Because the positions of each of the encapsulated protein elements, i.e. groups 120, is known to the precision of their micro lithographic deposition on the slide 101 , the automated analysis system is configured to move to the region of interest and begins its analysis. For example, the groups 120 of encapsulated proteins may be deposited in a series of increasing concentrations following some type of scale (linear, logarithmic, etc.). By measuring the integrated optical density for each of the groups 120 of encapsulated proteins in the series, the automated analysis system may make a high-resolution determination of the stain uptake for that protein. When compared to the previously-analyzed effects of various stain uptakes, the automated analysis system can determine whether that stain batch is suitable for medical or biological analysis and also which, if any, of the stains is beginning to fail.

Furthermore, if the groups 120 of encapsulated proteins are in the form of optical resolution test targets, the automated analysis system may make a simultaneous evaluation of its optical integrity by comparing this target to previously-evaluated targets. Alternatively, the test slide 100 could simply hold a standard resolution test target along with the encapsulated proteins.

In summary, the test slide with encapsulated protein test specimen provides a number of advantages including the following. (1 ) Predictable characteristics: The synthetically encapsulated proteins can be manufactured in a variety of ways, with high tolerances, to arrive at test specimens that supply predictable and quantifiable results when stained. The ability to be able to predict these results under a variety of conditions allows the automated image analysis system to perform both self-checks and preparation checks to ensure the integrity of its performance. The naturally encapsulated proteins can supply a similarly predictable test specimen if some type of cellular monotype is used and harvested correctly, as described above. (2) Standard format: the use of microscope slides (or equivalent) in order to create test specimens allows their introduction into the chain of specimen preparation and analysis without interrupting the analysis chain or unduly affecting the outcomes. (3) Increased dynamic range: In the case of a synthetically encapsulated protein set, the use of a variety of protein densities and/or over-coating procedures permits the creation of a test specimen slide 100 whose sensitivity to stain variations is much greater than that of a typical cytological or histological specimen. This allows better resolution of subtle distinctions between staining results in the test slide specimen 100 and is used to both establish the suitability of the staining procedure and estimate its time to failure.

Advantageously, the test slide 100 with encapsulated protein according to the present invention provides a vehicle or mechanism that fits into the natural workflow of an automated system, for example, an automated image analysis system as described above.

The present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. Certain adaptations and modifications of the invention will be obvious to those skilled in the art. Therefore, the presently discussed embodiments are considered to be illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims

WHAT IS CLAIMED IS:

1. A test slide apparatus for assessing stain quality of a biological specimen in an automated analysis system, said apparatus comprising:

(a) a slide carrier;

(b) a layer applied to a surface of said slide carrier, said layer forming a micro-porous surface;

(c) a stain-reactive element, said stain-reactive element being deposited in at least some of the micro-pores in the layer on the surface of said slide carrier.

2. The test slide as claimed in claim 1 , further including an overcoat layer applied over the micro-porous surface of said layer.

3. The test slide as claimed in claim 1 or 2, wherein said stain-reactive element comprises one or more encapsulated proteins deposited on the micro- porous surface of said layer.

4. The test slide as claimed in claim 3, wherein said stain-reactive elements are deposited in predetermined groups in the micro-porous layer.

5. The test slide as claimed in claim 4, wherein said encapsulated proteins comprise one or more proteins selected from the group of proteins including synthetic DNA, synthetic RNA, keratin, or albumin.

6. The test slide as claimed in ciaim 4, wherein said encapsulated proteins comprise a naturally encapsulated protein cultured from an immortal cell line.

7. A method for assessing operation of an automated analysis system, the method comprises the steps of:

(a) providing a test slide for the automated analysis system, said test slide comprising: (i) a slide carrier;

(ii) a layer applied to a surface of said slide carrier, said layer forming a micro-porous surface; (iii) a stain-reactive element, said stain-reactive element being deposited in at least some of the micro-pores in the layer on the surface of said slide carrier;

(b) applying said test slide to the automated analysis system and operating the automated system to perform selected processing procedures on said test slide to produce one or more test results for said test slide;

(c) recording the test results for said test slide so that said stain- reactive element provides a predictable response for the operation of the automated analysis system.

8. The method for assessing operation of an automated analysis system as claimed in claim 7, wherein said stain-reactive element comprises one or more encapsulated proteins deposited on the micro-porous surface of said test slide.

9. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said encapsulated proteins comprise one or more proteins selected from the group of proteins including synthetic DNA, synthetic RNA, keratin, or albumin.

10. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said encapsulated proteins comprise a naturally encapsulated protein cultured from an immortal cell line.

11. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said selected processing procedure performed by the automated analysis system comprises quantifying the density of the cells encapsulated on said test slide.

12. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said selected processing procedure per ormed by the automated analysis system comprises applying a segmentation operation to the encapsulated protein cells deposited on said test slide.

13. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said selected processing procedure performed by the automated analysis system comprises applying a feature extraction operation to the encapsulated protein cells deposited on said test slide.

14. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said selected processing procedure performed by the automated analysis system comprises applying an image verification operation to the encapsulated protein cells deposited on said test slide.

15. The method for assessing the operation of an automated analysis system as claimed in claim 8, wherein said selected processing procedure performed by the automated analysis system comprises applying a positioning accuracy operation to measure the accuracy of the placement of the encapsulated protein cells on said test slide.