Sequential photobleaching of fluorochromes for polychromatic slide‐based cytometry
Open Access
- 14 February 2006
- journal article
- research article
- Published by Wiley in Cytometry Part A
- Vol. 69A (3) , 139-141
- https://doi.org/10.1002/cyto.a.20227
Abstract
Background: Slide‐based cytometry is a key technology for polychromatic cytomic investigations. Here we exploit the relocalization and merge feature of Laser Scanning Cytometry for distinguishing fluorochromes of comparable emission spectra but different photostabilities.Methods: Blood specimens were stained with the fluorochrome pairs: FITC/ALEXA488, PE/ALEXA532, or APC/ALEXA633. Bleaching was performed by repeated laser excitation.Results: Since ALEXA dyes are photostable as compared to the conventional fluorochromes FITC, PE, and APC, a differentiation within one fluorochrome pair is possible.Conclusion: The sequential photobleaching method results in an increased information density on a single cell level and represents an important component to perform polychromatic cytometry. © 2006 International Society for Analytical CytologyKeywords
This publication has 9 references indexed in Scilit:
- Exploring molecular networks directly in the cellCytometry Part A, 2006
- Iterative restaining as a pivotal tool for n‐color immunophenotyping by slide‐based cytometryCytometry Part A, 2006
- Polychromatic (eight‐color) slide‐based cytometry for the phenotyping of leukocyte, NK, and NKT subsetsCytometry Part A, 2005
- Human cytome project, cytomics, and systems biology: The incentive for new horizons in cytometryCytometry Part A, 2005
- Application of spectral imaging microscopy in cytomics and fluorescence resonance energy transfer (FRET) analysisCytometry Part A, 2004
- Quantitative histology by multicolor slide‐based cytometryCytometry Part A, 2004
- Six and more color immunophenotyping on the slide by laser scanning cytometry (LSC)Published by SPIE-Intl Soc Optical Eng ,2003
- Near‐infrared dyes for six‐color immunophenotyping by laser scanning cytometryCytometry, 2002
- Alexa Dyes, a Series of New Fluorescent Dyes that Yield Exceptionally Bright, Photostable ConjugatesJournal of Histochemistry & Cytochemistry, 1999