NDLI: Fast fluorescence lifetime imaging of calcium in living cells.

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Fast fluorescence lifetime imaging of calcium in living cells.

Content Provider	Semantic Scholar
Author	Agronskaia, Alexandra V. Tertoolen, Leon L. Gerritsen, Hans C.
Copyright Year	2004
Abstract	A fast fluorescence lifetime imaging (FLIM) system is developed that can acquire images at a rate of hundreds of frames per second. The FLIM system is based on a wide-field microscope equipped with a time-gated intensified CCD detector and a pulsed laser. The time-gated detector acquires the signals from two time gates simultaneously and is therefore insensitive to movements of the specimen and photo-bleaching. The system is well suited for quantitative biological FLIM experiments and its performance is evaluated in calcium imaging experiments on beating neonatal rat myocytes. Several calcium sensitive dyes are characterized and tested for their suitability for fast FLIM experiments: Oregon Green Bapta-1 (OGB1), Oregon Green Bapta-2 (OGB2), and Oregon Green Bapta-5N (OGB5N). Overall the sensitivity range of these dyes is shifted to low calcium concentrations when used as lifetime dyes. OGB1 and OGB2 behave very similarly and can be used for FLIM-based calcium imaging in the range 1 to approximately 500 nM and OGB5N can be used up to 3 microM. The fast FLIM experiments on the myocytes could be carried out at a 100-Hz frame rate. During the beating of the myocytes a lifetime change of about 20% is observed. From the lifetime images a rest calcium level of about 65 nM is found.
Starting Page	4741
Ending Page	4751
Page Count	11
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://www.bioee.ee.columbia.edu/courses/upload/Bibliography/agronskaia2004.pdf
PubMed reference number	15568944v1
Volume Number	9
Issue Number	6
Journal	Journal of biomedical optics
Language	English
Access Restriction	Open
Subject Keyword	Benzalkonium Chloride 1 MG/ML Topical Foam [Lite 'N Foamy] Buffers CDISC SEND Biospecimens Terminology Calcium measurement Calibration Cyanobacteria Detectors Dyes Excitation Fluorescence Frame (physical object) Indocyanine Green Ions Magnetic Resonance Imaging Microscope Device Component Movement Muscle Cells Photobleaching Preparation
Content Type	Text
Resource Type	Article

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