Confocal microscopic imaging of fast UV-laser photolysis of caged compounds.
Using a pulsed UV laser in a confocal scanning microscope, we present a relatively cheap, accurate and efficient method for fast UV laser flash photolysis of caged molecules in two-dimensional cultured neurons. The laser light is introduced through the imaging optics, can be localized by a parallel red laser and can photolyse a sphere of less than 1 microm2, and evoke local fluorescence changes in the imaged neurons. Caged glutamate and caged fluorescein are used to illustrate a disparity between spines and their parent dendrites at a sub-micron resolution.
Korkotian
Eduard
E
Department of Neurobiology, The Weizmann Institute, Rehovot 76100, Israel.
Oron
Dan
D
Silberberg
Yaron
Y
Segal
Menahem
M
D002478:Cells, Cultured; D003712:Dendrites / Q000378:metabolism; D019793:Fluorescein / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000378:metabolism; D005971:Glutamates / Q000493:pharmacokinetics; D007834:Lasers; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D009474:Neurons / Q000502:physiology; D010782:Photolysis; D014466:Ultraviolet Rays
D005456:Fluorescent Dyes; D005971:Glutamates; C082764:alpha-(4,5-dimethoxy-2-nitrobenzyl) glutamate; D019793:Fluorescein