Image cytometry of estrogen receptors in breast carcinomas

A significant level of estrogen receptors (ER) in breast cancer cells is an indication of tumor differentiation and suggests that a homeostatic control of cell growth may persist in these cancers. In medical practice, the Dextran-coated charcoal assays (DCCA) are still the most frequently used test to characterize patients having ER-positive malignant breast tumors and for whom hormonal therapy is justified. Nevertheless, this routine biochemical technique is not satisfactory because it is a broad method unsuitable for revealing receptor tissue heterogeneity. However, immunocytochemical labeling, such as the ER-ICA method, which involves a monoclonal antibody linked to peroxidase, is a specific reaction for this purpose but which until now was not quantitative. The present study uses an original cell preparation technique combining the PAP reaction with toluidine blue counterstain for image analysis on the SAMBA system. Special software has been developed for the quantitative analysis of immunocytochemistry in cancers. Results obtained showed a high correlation between the DCCA values and the score derived from the mean ER concentration per positive tumor cell and the labeling index. In addition, intracell and intratumor heterogeneity can be displayed according to several parameters and were shown to vary according to tumor and to antiestrogen (Tamoxifen) presurgical therapy.     

A double scanning microphotometer for image analysis: hardware, software and biomedical applications.

A new image processing system designed for densitometry and pattern analysis of microscopic specimens is described with special regard to the hardware, the software and the biologic applications. The data acquisition procedure involves the combination between the scanning of the preparation by means of a motorized stage and the scanning of successive fields by a mechanical device. The signal provided by the photomultiplier is converted into digital values which are directed to an on-line computer. The data processing is based on a one-pass computation involving automata theory and therefore it avoids the storage of the image in the computer memory. In so doing, an entire and continuous image of the whole preparation can be processed at the highest magnification of the microscope whatever the size of the analyzed specimen may be. A biologic application of the system is reported and concerns the automatic identification and counting of cells in the various phases of the mitotic cycle.     

The HOME microscope workstation. A new tool for cervical cancer screening.

The Highly Optimized Microscope Environment (HOME) is a computerized microscope design for assisting pathologists and cytotechnologists in routine clinical tasks. The prototype system consists of an IBM PC-compatible computer and a light microscope in which a built-in high-resolution computer display image is super-imposed on the optical image of the specimen. Also, an encoding stage and objective turret encoder are used to provide continuous monitoring of the stage coordinates and microscope magnification to the computer. This allows any position on the stage to be uniquely defined. Software, written in C language and running under the MS-DOS/MS-Windows environment, is controlled by means of a mouse-driven cursor. A specific application has been developed for cervical cancer screening, taking into account the needs and constraints of microscopists performing this task. Informatics tools offered by the HOME system provide them with precise flagging and relocation of objects on the slide, control of the scanning pathway, and ability to write and print the report directly through the microscope. The computer files generated by microscopic examination are stored and contain information available for quality control assessment and laboratory management.     

Un nouveau système d'analyse densitométrique et morphologique des préparations microscopiques

Résumé La reconnaissance et le comptage des cellules dans les différentes phases du cycle mitotique ont été réalisés à l'aide d'un nouveau système d'analyse densitométrique et morphologique des préparations microscopiques. Le processus d'analyse est entièrement automatique et consiste à balayer l'image de champs successifs de la préparation avec une définition égale au pouvoir de résolution du microscope. L'alternance entre le balayage de l'image et les déplacements de la préparation est commandée et contrôlée par une unité électronique programmable organisée autour d'un microprocesseur. Le traitement des données est réalisé par un ordinateur sans stockage de l'image en unité centrale; il n'est donc pas limité par le nombre des données analysées. Les résultats préliminaires de l'analyse densitométrique et morphologique de cellules embryonnaires télencéphaliques montrent que le système utilisé permet de reconnaître et de compter les cellules en mitose, les cellules en phases G₁, S et G₂ et les cellules qui ne sont pas engagées dans un cycle mitotique (phase G₀ sensu largo). Ces données permettent d'établir les paramètres de la cinétique de prolifération dans la population étudiée, et notamment l'indice mitotique et le coefficient de prolifération, à partir d'une seule préparation histologique.

---

A new image-processing system designed for densitometry and pattern analysis of microscopic specimen. Application to the automated recognition and counting of cells in the various phases of the mitotic cycle

Summary A new image analysing system, designed for microphotometric measurement and pattern recognition has been applied in the discrimination of cells from the various phases of the mitotic cycle. The data acquisition procedure is controlled by a programmable electronic unit and involves the combination of the shifting of the microscope moving stages and the scanning of the successive fields by a mechanical device. The data processing is achieved by a computer. The preliminary results we obtained have shown that such a system allows the automatic recognition and counting of the M, G₁, S and G₂ cells as also the G₀ resting cells. The most useful parameters of the cell proliferation kinetics are thus obtained from a single specimen of a cell population.

---

---

Ce travail a bénéficié de l'aide du CNRS (ATP N^o A 6551799 et ATP N^o 1751) et de l'Inserm (AT N^o 74 14 2036). Brevet Anvar 11 462     

SAMBA : a cytophotometer coupled to a computer for the purpose of the automatic study of cell kinetics and the recognition of cell types]

The system SAMBA has been designed for automatic analysis of biological images, at the cellular or subcellular level. The examination is performed at the maximum resolution power of the microscope. It enables to discriminate by means of a pool of morphological and densitometrical parameters, between cells in the different phases of the cell cycle, or between cells belonging to various types in heterogeneous populations. Other recognition programs are at present beeing in progress, in order to promote the use of SAMBA in other fields of fundamental research and clinical application. Owing to the minicomputer now in use, SAMBA is an autonomous system, capable of beeing routinely used in diagnostic centers.