Fast Dual-excitation Ratiometry with Light-emitting Diodes and High-speed Liquid Crystal Shutters.
From: Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
Biochemical and biophysical research communications
- Publish Date: Feb 2006
- ISSN: 0006-291X
- Volume: 340
- Issue: 1
- Pages: 250-5
- Medium: Print
- Language: English
- Citation (JAMA): Fukano Takashi, Shimozono Satoshi, Miyawaki Atsushi, et al. Fast Dual-excitation Ratiometry with Light-emitting Diodes and High-speed Liquid Crystal Shutters.. Biochem. Biophys. Res. Commun. Feb 2006;340:250-5
Abstract
Dual-excitation ratiometric dyes permit quantitative measurements of Ca2+ concentrations ([Ca2+]s), by minimizing the effects of several artifacts that are unrelated to changes in [Ca2+]. These dyes are excited at two different wavelengths, and the resultant fluorescence intensities are measured sequentially. Therefore, it is difficult to follow fast [Ca2+] dynamics or [Ca2+] changes in highly motile cell samples. To overcome this problem, we have developed a new dual-excitation ratiometry system that employs two high-power light-emitting diodes (LEDs), two high-speed liquid crystal shutters, and a CCD camera. The open/close operation of the two shutters is synchronized with the on/off switching of the two LEDs. This system increases the rate at which ratio measurements are made to 1 kHz, and provides ratio images at 10-100 Hz depending on the signal intensity. We demonstrate the effectiveness of this system by monitoring changes in [Ca2+] in cardiac muscle cells loaded with Fura-2.
Mesh Headings (Keywords): Animals, Calcium, Cells, Cultured, Equipment Design, Equipment Failure Analysis, Fura-2, Image Enhancement, Lighting, Liquid Crystals, Microscopy, Fluorescence, Multiphoton, Myocytes, Cardiac, Rats, Semiconductors, Signal Processing, Computer-Assisted
Check for Full Text / PubMed Unique Identifier (PMID): 16360639
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