Abstract
We have constructed a modular instrument to measure intracellular [Ca2+] ([Ca2+]i) in single isolated cells while simultaneously imposing step changes in [Ca2+]i using "caged Ca2+". By combining the outputs of a xenon arc lamp with a frequency-tripled (Nd: YAG) laser, the instrument can operate with low maintained illumination to measure [Ca2+]i using a ratiometric Ca2+-sensitive fluorophore and also activate the release of Ca2+ from a caged-Ca2+ compound with a high energy pulse of ultraviolet light. This instrument is simple to assemble, introduces little electrical noise, provides a wide range of illumination power, produces only moderate photobleaching of the Ca2+ indicator and can be readily adapted to diverse cellular preparations. We demonstrate the use of this system to measure step changes in [Ca2+]i in adult rat ventricular myocytes and a human embryonic kidney cell line (293 cells) in culture.
Original language | English |
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Pages (from-to) | 169-177 |
Number of pages | 9 |
Journal | Pflugers Archiv European Journal of Physiology |
Volume | 427 |
Issue number | 1-2 |
DOIs | |
State | Published - May 1994 |
Keywords
- 293 cells
- Cardiac cells
- DM-nitrophen
- Flash photolysis
- Indo-1
- Nd: YAG laser
ASJC Scopus subject areas
- Physiology
- Clinical Biochemistry
- Physiology (medical)