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A simple cell labeling technique by means of lectins linked to fluorochromes for the detection of cells on tissue sections
Author(s) -
AlonsoVarona Ana,
Calle Yolanda,
Palomares Teodoro,
Castro Begoña,
BarberáGuillem Emilio
Publication year - 1995
Publication title -
biology of the cell
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.543
H-Index - 85
eISSN - 1768-322X
pISSN - 0248-4900
DOI - 10.1016/0248-4900(96)89935-7
Subject(s) - biology , wheat germ agglutinin , lewis lung carcinoma , lectin , cell , microbiology and biotechnology , cell culture , fluorescein isothiocyanate , pathology , metastasis , biochemistry , cancer , fluorescence , genetics , quantum mechanics , medicine , physics
Summary— We designed a protocol for cell labeling with the lectin wheat germ agglutinin (WGA) linked to the fluorochrome tetramethyl‐rhodamine isothiocyanate (TRITC) for effective detection of the B16F10 melanoma and Lewis lung carcinoma (LLc) cells on pulmonary histological sections from C57BL6; mice. We have also determined a suitable concentration of WGA‐TRITC (10 μg/ml), which leads to a very intense and homogeneous labeling of the cells, as it avoids cell clumping due to the presence of the lectin WGA. In order to determine to what extent the method affects these tumor cells, we have studied some important aspects related to their metastatic behavior, taking into account three parameters: a) viability and rate of proliferation of the cells cultured in vitro ; b) percentage of animals C57BL6 mice) bearing metastasis 15 days after intravenous inoculation with 10 5 B16F10 or LLc cells; and c) pattern of distribution of tumor foci in lung. There were no significant differences in these three parameters between the WGA‐TRITC labeled‐cells compared to the cultures of non‐labeled cells in either of the cell lines (B16F10, LLc). Thus, we conclude that B16F10 and LLc tumor cells can be labeled following the protocol set‐up in our study, as it allows these cells to be neatly identified on tissue sections and it causes no important physiological changes in the cells, with regard to metastatic behavior. These points make this technique very suitable for the detection of B16F10 and LLc cells on histological sections in studying their behavior during the first stages of the metastatic process.