SPECIFIC APPLICATIONS OF CYTOKERATIN ANTIBODIES
DIRECTLY LABELED CYTOKERATIN ANTIBODIES FOR
APPLICATION IN FLOW CYTOMETRY
To those investigators and clinicians dealing with DNA
flow cytometric analysis of patient tumors it may be a daily experience that such analyses
of carcinomas are in many instances disturbed or made impossible by the presence of
stromal and inflammatory cells in the tumor cell suspensions. The variability of these
admixtures makes it often difficult to determine the proliferation capacity or even to
estimate the ploidy of a certain tumor. These parameters have, however, been shown to be
of prognostic significance for several types of tumors.
In order to exclude the non-epithelial cell types from
such analyses we have developed a two-parameter method using cytokeratin antibodies. For
this purpose single cell suspensions from fresh frozen tissues are fixed in 70 % ethanol
and stained with FITC-conjugated cytokeratin antibodies. Thereafter DNA is labelled with
propidium iodide. The epithelial (cytokeratin positive) cells can now be separated from
non-epithelial (cytokeratin negative) cells
in a two-parameter analysis.
Recently we have developed a procedure to prepare and
analyse paraffin embedded tissues in such a two parameter cytokeratin/DNA flow cytometric
assay.
