Spectral Imaging of MC540 During Murine and Human Colon Carcinoma Cell Differentiation

SUMMARY We studied the staining pattern of merocyanine 540 (MC540) by spectral imaging of murine CT26 and human HT29 colon carcinoma cells incubated with the dye MC540. This dye, usually considered a potential membrane probe, localized mainly in the cytoplasmic vesicles of the colon carcinoma cells....

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Published in:The journal of histochemistry and cytochemistry 2001-02, Vol.49 (2), p.147-153
Main Authors: Siboni, Galit, Rothmann, Chana, Ehrenberg, Benjamin, Malik, Zvi
Format: Article
Language:English
Subjects:
Alkaline Phosphatase - metabolism
Animals
Butyrates
Cell Differentiation
Cell Division
Colonic Neoplasms - pathology
Flow Cytometry
Fluorescent Dyes
Humans
Hydrogen-Ion Concentration
Mice
Pyrimidinones
Spectrometry, Fluorescence
Subcellular Fractions - metabolism
Tumor Cells, Cultured
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Summary:SUMMARY We studied the staining pattern of merocyanine 540 (MC540) by spectral imaging of murine CT26 and human HT29 colon carcinoma cells incubated with the dye MC540. This dye, usually considered a potential membrane probe, localized mainly in the cytoplasmic vesicles of the colon carcinoma cells. However, in cells incubated in an environment similar to that of a tumor (pH 6.7), high fluorescence was detected in the nuclear membrane and nucleoli. Under these acidic conditions, resembling the Krebs effect, a population of CT26 cells displayed fluorescent plasma membranes. In differentiating cells, exhibiting cell cycle arrest at G1-phase and an elevated level of alkaline phosphatase, MC540 fluorescence was confined to cytoplasmic vesicles and was not detected in the plasma membrane or in the nucleoli. Cell sorting analysis of both cell types at pH 5.0 revealed higher fluorescence intensity in proliferating cells compared to differentiating cells. The fluorescence intensity of MC540-stained cells reached a maximum at pH 5.0, although the fluorescence of MC540 dye was maximal at pH 7.2. This phenomenon may result from increased binding of MC540 monomers to the cells because disaggregation of the dye with Triton X-100 produced similar results. We conclude that nucleolar localization of MC540 and an elevated fluorescence intensity can be used as indicators for proliferating cells in the characteristically acidic tumor environment. (J Histochem Cytochem 49:147–153, 2001)
ISSN:0022-1554
1551-5044
DOI:10.1177/002215540104900202