Measurement Conditions for Flow Cytometry Analyses of Cell Lines from Urological Carcinomas |
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Authors: | Angelika?T?lle Ziyad?Abdallah Klaus?Jung Hans?B?umler |
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Institution: | 1.Department of Urology,Charité–Universit?tsmedizin Berlin,Berlin,Germany;2.Institute of Transfusion Medicine,Charité—Universit?tsmedizin Berlin,Berlin,Germany;3.Berlin Institute for Urologic Research,Berlin,Germany;4.Berlin Brandenburg Center for Regenerative Therapies,Berlin,Germany |
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Abstract: | Prerequisites for successful flow cytometry investigations are specific antibodies labeled with appropriate fluorochromes
and negligible autofluorescence of the untreated cells at the wavelength of interest. The aim of this study was (a) to characterize
frequently used urological carcinoma cell lines with regard to their autofluorescence properties, (b) to demonstrate the autofluorescence
as a serious interfering factor on FACS analysis of urological carcinoma cell lines and (c) to suggest an alternative to avoid
interfering autofluorescence. Twenty-one cell lines originating from prostate carcinoma, renal cell carcinoma and bladder
cancer were included in this study. The various cell lines were read on a flow cytometer in comparison to human erythrocytes
as cells with low fluorescence intensity. Urological cell lines show a high autofluorescence when flow cytometry analyses
are performed at the frequently used excitation wavelengths at 405 and 488 nm. At excitation wavelength of 633 nm, this problem
was reduced and most of the cell lines (14/21) were without autofluorescence at the emission wavelength of 785 nm. In addition,
with a spectrofluorometer three exemplary cell lysates were investigated. The above observations were confirmed. The dye APC-Cy7
is one suitable fluorochrome for successful investigation under these measurement conditions. |
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