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Free keywords:
SUPERRESOLUTION MICROSCOPY; SCMOS CAMERA; LIVE CELLS; LOCALIZATION;
MOLECULES; TRANSPORT; SYNAPSES; PROTEINS; TRACKINGAnatomy & Morphology; Life Sciences & Biomedicine - Other Topics;
Microscopy; fluorescence microscopy; CCD; CMOS; time-resolved imaging;
Abstract:
The plethora of available scientific cameras of different types challenges the biologically oriented experimenter when picking the appropriate camera for his experiment. In this study, we chose to investigate camera performances in a typical nonsingle molecule situation in life sciences, that is, quantitative measurements of fluorescence intensity changes from video data with typically skewed intensity distributions. Here, intensity profile dynamics of pH-sensors upon triggered changes of pH-environments in living cells served as a model system. The following camera types were tested: sCMOS, CCD (scientific and nonscientific) and EM-CCD (back-and front-illuminated). We found that although the EM-CCD cameras achieved the best absolute spatial SNR (signal-to-noise ratio) values, the sCMOS was at least of equal performance when the spatial SNR was related to the effective dynamic range, and it was superior in terms of temporal SNR. In the measurements of triggered intensity changes, the sCMOS camera had the advantage that it used the smallest fraction of its dynamic range when depicting intensity changes, and thus featured the best SNR at full usage of its dynamic range. (C) 2013 Wiley Periodicals, Inc.
