Cytophotometry, or image analysis, is conceptually similar to flow cytometery but requires a different preparations of cells for the measurements. The principle of this technique is the measurement of the optical density of cells in histological section or, even better, of whole cells spread in histological section or, even better, of whole cells spread on the glass.
Compared with flow cytometry, the technique is much slower, and fewer objects can be measured. The main advantage of this approach is the ability to identify a measured object microscopically and to perform the measurements separately on different cell populations and tissue components.
The technique is uniquely suited for the measurements of distinct tumor cell populations. Recent developments of computerized tissue reconstruction programs have made it possible to perform the quantitations on histologic sections.
The information provided is somewhat similar to data gained by flow cytometry. In pathology including that of bone tumors, this technique is most frequently used to generate DNA ploidy distribution patterns and to perform cell-cycle analysis. For this type of analysis, the cells are typically stained with Feulgen reaction.
As with flow cytometry, any cellular component that can be visualized directly by an appropriate color reaction or with the use of antibodies can be quantitated for investigative purposes. In addition, several other parameters, such as nuclear size, shape, volume, and chromatin texture, can be measured. In summary, the main advantage of this technique is its ability to verify the measured objects microscopically in standard cytologic and histologic preparations.
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