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9 March 2020 Multi-target immunofluorescence using spectral FLIM-FRET for separation of undesirable antibody cross-labeling and autofluorescence (Conference Presentation)
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Abstract
To overcome limitations of indirect immunofluorescence, a new method is presented to employ the ostensible disadvantage of cross-labeling secondary antibodies by separation of the fluorescence signals via spectral FLIM-FRET. The undesirable cross-labeling among secondary antibodies leads to the generation of new characteristic FRET emission spectra including a change in the donor lifetime. We used a spectrally resolved FLIM detection system with pulse interleaved excitation. The combined spectral FLIM-FRET and pattern-matching analysis forms an excellent tool for use in indirect immunofluorescence that overcomes the undesirable effect of secondary antibody cross-labeling by assigning separate color channels to cross-labeled fluorescent antibodies.
Conference Presentation
© (2020) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Sumeet Rohilla, Benedikt Kraemer, Felix Koberling, Uwe Ortmann, Ingo Gregor, Andreas C. Hocke, and Rainer Erdmann "Multi-target immunofluorescence using spectral FLIM-FRET for separation of undesirable antibody cross-labeling and autofluorescence (Conference Presentation)", Proc. SPIE 11244, Multiphoton Microscopy in the Biomedical Sciences XX, 1124418 (9 March 2020); https://doi.org/10.1117/12.2542413
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