CCQM-P123 number and geometric property of cells adhered to a solid substrate

Le résumé fourni par la source

Main text Several scientific fields, such as cytology, biochemistry, molecular biology and genetics have cells as their direct topic of study. The cell is a fundamental unit of highly technological systems, employed, for example, in the development of new chemicals and drugs, in the evaluation of compound toxicity, in novel approaches to health, such as regenerative medicine. High content screening technologies are cellular imaging-based assays. In this framework, cell density, cell confluency and cell size are measurands that can describe a biological effect of a molecule under examination. The CCQM-P123 study aimed to demonstrate capability of participants in quantifying 4 measurands using one reference material: number of adherent cells on a planar surface, cell density in the range 0 to 5000 cells/mm2, cell confluency in the range 0 % to 100 % and average cell area. Nine Institutions participated in the study. Reference materials for three levels of cell density and cell confluency were prepared for fluorescence microscopy and circulated among participants. Each reference material was a commercial imaging dish having 400 squares and seeded with human cells, fixed and stained for nucleus and for whole cell. On each dish 4 squares were selected for the analysis as "measurement standards". The use of a calibrated ruler to define the area of measurement assured the traceability to the International System of Units (SI). Results for the 4 measurands showed an overall good agreement between the laboratories. The study allowed fundamental considerations in the frame of a cell-based measurement comparison, in terms of: traceability, comparability of results and complexity of the study protocol. The study has been a preparatory exercise for a key comparison on at least one of the 3 measurements claims (cell density, cell confluency and cell area) related to adhered cells features such as cell proliferation, morphology and size. To reach the main text of this paper, click on Final Report . The final report has been peer-reviewed and approved for publication by the CCQM.

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