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Content archived on 2024-04-19

The prototype reference standard slides for quantitative cytometry of nuclear DNA content

Objective



Recent advances in cytometry techniques have led to attempt to develop diagnostic information and to classify tumours based on their basic biologic properties such as the DNA content of cells, incorrectly called "ploidy".

It has been firmly established that the determination of nuclear DNA content in human tumours provides important diagnostic and prognostic information that is useful in selecting the most appropriate therapeutic strategy for managing the disease. Image cytometry (ICM) techniques have been successfully applied to the analysis of tumour cell DNA content in breast cancers. At present numerous pathological laboratories, cancer prevention centres, hospitals and private laboratories use DNA content information as a diagnostic and prognostic tool.

Briefly, the full procedure involved in DNA content analysis by using ICM methods involves five successive steps : the cytological preparation of the clinical specimen (sampling, fixation and Feulgen staining), the acquisition of the image, the analysis of the image, the nuclear DNA densitometric featuring and the interpretation of DNA measurements. It appears clearly that the result of a specimen evaluation is dependent on conditions of specimen preparation, and image analysis instrumentation and procedures.

The implementation of this new methodology in the daily clinical practice of histopathological diagnosis raises major questions about the definition of detailed guide-lines for completion of the analysis procedure involved. Consequently, requirements for rigorous technical quality of cytological preparation, of instrumentation, of standards for normalisation of densitometric measurements to make reliable quantitation and for rules to extract accurate information from measurements must be defined in detail.

The project will define prototypes of reference standard slides to be used for calibration of ICM instrumentation and for normalisation of nuclear DNA measurements.
Moreover, it will determine specific guide-lines to be used for accurate ICM DNA quantisation.
Finally, it will lead to a harmonization of medical knowledge and rules of decision making in the field of breast cancer.

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Coordinator

Université de Grenoble I (Université Joseph Fourier)
EU contribution
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Address
Domaine de la Merci
38700 La Tronche
France

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Participants (4)

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