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Proceedings Paper

Screening of cardiomyocyte fluorescence during cell contraction by multi-dimensional TCSPC
Author(s): D. Chorvat Jr.; S. Abdulla; F. Elzwiei; A. Mateasik; A. Chorvatova
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Paper Abstract

Autofluorescence is one of the most versatile non-invasive tools for mapping the metabolic state of living tissues, such as the heart. We present a new approach to the investigation of changes in endogenous fluorescence during cardiomyocyte contraction - by spectrally-resolved, time correlated, single photon counting (TCSPC). Cell contraction is stimulated by external platinum electrodes, incorporated in a home-made bath and triggered by a pulse generator at a frequency of 0.5 Hz (to stabilize sarcoplasmic reticulum loading), or 5 Hz (the rat heart rate). Cell illumination by the laser is synchronized with cell contraction, using TTL logic pulses operated by a stimulator and delayed to study mitochondrial metabolism at maximum contraction (10-110 ms) and/or at steady state (1000-1100 ms at 0.5 Hz). To test the setup, we recorded calcium transients in cells loaded with the Fluo-3 fluorescent probe (excited by 475 nm pulsed picosecond diode laser). We then evaluated recordings of flavin AF (excited by 438 nm pulsed laser) at room and physiological temperatures. Application of the presented approach will shed new insight into metabolic changes in living, contracting myocytes and, therefore, regulation of excitation-contraction coupling and/or ionic homeostasis and, thus, heart excitability.

Paper Details

Date Published: 15 February 2008
PDF: 12 pages
Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 686029 (15 February 2008); doi: 10.1117/12.784756
Show Author Affiliations
D. Chorvat Jr., International Laser Ctr. (Slovak Republic)
S. Abdulla, Univ. of Montréal (Canada)
F. Elzwiei, Univ. of Montréal (Canada)
A. Mateasik, International Laser Ctr. (Slovak Republic)
A. Chorvatova, Univ. of Montréal (Canada)

Published in SPIE Proceedings Vol. 6860:
Multiphoton Microscopy in the Biomedical Sciences VIII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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