Flash photolysis using a light emitting diode: an efficient, compact, and affordable solution.
Flash photolysis has become an essential technique for dynamic investigations of living cells and tissues. This approach offers several advantages for instantly changing the concentration of bioactive compounds outside and inside living cells with high spatial resolution. Light sources for photolysis need to deliver pulses of high intensity light in the near UV range (300-380 nm), to photoactivate a sufficient amount of molecules in a short time. UV lasers are often required as the light source, making flash photolysis a costly approach. Here we describe the use of a high power 365 nm light emitting diode (UV LED) coupled to an optical fiber to precisely deliver the light to the sample. The ability of the UV LED light source to photoactivate several caged compounds (CMNB-fluorescein, MNI-glutamate, NP-EGTA, DMNPE-ATP) as well as to evoke the associated cellular Ca(2+) responses is demonstrated in both neurons and astrocytes. This report shows that UV LEDs are an efficient light source for flash photolysis and represent an alternative to UV lasers for many applications. A compact, powerful, and low-cost system is described in detail.
Bernardinelli
Yann
Y
Department of Physiology, University of Lausanne, Switzerland.
Haeberli
Christian
C
Chatton
Jean-Yves
JY
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D005336:Fiber Optic Technology; D019793:Fluorescein; D005971:Glutamates / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D008027:Light; D051379:Mice; D055100:Optical Fibers; D010782:Photolysis; D014466:Ultraviolet Rays
C440256:4-methoxy-7-nitroindolinyl-glutamate; D005971:Glutamates; D007211:Indoles; C085112:2-nitrophenyl-EGTA; D004533:Egtazic Acid; D002118:Calcium; D019793:Fluorescein