Flow cytometry and scrape-loading/dye transfer as a rapid quantitative measure of intercellular communication in vitro.
We describe two flow cytometric assays performed on populations of cells which have been stained with various fluorescent tracer molecules by the scrape-loading technique. One assay uses a simple one-color analysis on a flow cytometer by quantitating the fluorescence intensity of scrape-loaded lucifer yellow CH (LY) in individual cells. The other assay utilizes a two-color analysis on a cell sorter whereby cells which are initially loaded (donors) are identified by their uptake of both rhodamine isothiocyanate-dextran and LY, whereas the recipients of dye transfer are identified as having LY only. Agents which have been shown to inhibit intercellular communication in other assays exhibit similar blocking activity in LY transfer and this is readily quantitated by flow cytometry. The two-color analysis has the added advantage of being able to identify both donors and recipients in a highly quantitative manner.
Kavanagh
T J
TJ
Department of Pathology SM-30, University of Washington, Seattle 98195.
Martin
G M
GM
el-Fouly
M H
MH
Trosko
J E
JE
Chang
C C
CC
Rabinovitch
P S
PS
D002450:Cell Communication; D002455:Cell Division; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes; D006652:Histological Techniques; D007546:Isoquinolines; D008168:Lung; D012235:Rhodamines
D005456:Fluorescent Dyes; D007546:Isoquinolines; D012235:Rhodamines; C017475:lucifer yellow; C027755:rhodamine isothiocyanate