Megapixel FLIM

Hauke Studier; Klaus Weisshart; Oliver Holub; Wolfgang Becker

doi:10.1117/12.2039454

28 February 2014 Megapixel FLIM

Hauke Studier, Klaus Weisshart, Oliver Holub, Wolfgang Becker

Proceedings Volume 8948, Multiphoton Microscopy in the Biomedical Sciences XIV; 89481K (2014) https://doi.org/10.1117/12.2039454
Event: SPIE BiOS, 2014, San Francisco, California, United States

Abstract

TCSPC (Time-Correlated Single-Photon Counting) FLIM data with megapixel resolution can be recorded by using bh TCSPC modules in combination with new 64 bit data acquisition software. The large memory space available in the 64 bit environment allows new FLIM procedures to be used. We demonstrate the performance for applications that require imaging of a large number of cells in a single field of view, for multi-wavelength FLIM, for spatial mosaic imaging, and for recording transient changes in the fluorescence decay after a stimulation of the sample. Image quality was further improved by integrating a parallel counter channel that bypasses the timing electronics of the TCSPC module. Photon numbers from this counter are not affected by dead-time effects. Lifetime images are built up by using intensity data from the parallel counter and fluorescence decay data from the TCSPC electronics.

Citation Download Citation

Hauke Studier, Klaus Weisshart, Oliver Holub, and Wolfgang Becker "Megapixel FLIM", Proc. SPIE 8948, Multiphoton Microscopy in the Biomedical Sciences XIV, 89481K (28 February 2014); https://doi.org/10.1117/12.2039454

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