Confocal image analysis of three‐dimensional intracellular protein distribution: Application to cyclin A distribution in lymphoid cells

Autor: Paul André Bryon, Richard Delorme, Martine French, Catherine Souchier
Rok vydání: 1994
Předmět:
Zdroj: Bioimaging. 2:69-77
ISSN: 1361-6374
0966-9051
DOI: 10.1002/1361-6374(199406)2:2<69::aid-bio1>3.3.co;2-5
Popis: This paper proposes to evaluate, in tonsil lymphoid cells, the intracellular distribution of cyclin A, a protein involved in the cell cycle regulation. Serial sections of anti-cyclin A labelled cells were optically acquired with a confocal laser scanning microscope. On these serial sections, three-dimensional image analysis was applied, essentially by extending previous methods from the two-dimensional to the three-dimensional field. Sieving techniques derived from mathematical morphology were used to extract cluster- and spot-shaped cyclin A structures. Subsequently, measurements were performed to estimate cell surface and volume, cyclin A contents, cyclin A patterns and cytoplasmic or nuclear cyclin A localization. Results were finally submitted to a principal component analysis which ordered the cells in such a way as to satisfy human visual perception. Cyclin A distribution appeared to vary greatly, and in a continuous way as no group was obviously shared. Some typical patterns, such as spotted cyclin A were observed. Confocal laser scanning microscopy and three-dimensional analysis appeared as a relevant method to quantitatively investigate the subcellular distribution of proteins with respect to the fundamental three spatial dimensions of cells.
Databáze: OpenAIRE
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