Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals

The flash photolysis of "caged" compounds is a powerful experimental technique for producing rapid changes in concentrations of bioactive signaling molecules. These caged compounds are inactive and become active when illuminated with ultraviolet light. This paper describes an inexpensive a...

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Publicado:	2011
Materias:	Cage compound Calcium signal Confocal microscopy Inositol 1,4,5-trisphosphate receptor Photolysis UV illumination Cage compounds Confocal fluorescence Confocal microscopes Experimental techniques Exposure-time Flash photolysis Intracellular calcium Olympus Rapid changes Signaling molecules Ultra-violet light Ultraviolet illumination UV illuminations Wide area Xenopus laevis oocytes Alcohols Calcium Enzyme activity Fluorescence Fluorescence microscopy Mercury (metal) Optical fibers Sugars Ultraviolet radiation Microscopes Xenopus laevis 2 nitrophenyl EGTA 2-nitrophenyl-EGTA calcium drug derivative egtazic acid inositol 1,4,5 trisphosphate animal article calcium signaling chemistry confocal microscopy equipment design instrumentation metabolism methodology oocyte photolysis physiology statistical model ultraviolet radiation Animals Calcium Signaling Egtazic Acid Equipment Design Inositol 1,4,5-Trisphosphate Linear Models Microscopy, Confocal Oocytes Ultraviolet Rays
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10833668_v16_n6_p_Sigaut http://hdl.handle.net/20.500.12110/paper_10833668_v16_n6_p_Sigaut
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_10833668_v16_n6_p_Sigaut2023-06-08T16:05:53Z Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals Cage compound Calcium signal Confocal microscopy Inositol 1,4,5-trisphosphate receptor Photolysis UV illumination Cage compounds Confocal fluorescence Confocal microscopes Experimental techniques Exposure-time Flash photolysis Inositol 1,4,5-trisphosphate receptor Intracellular calcium Olympus Rapid changes Signaling molecules Ultra-violet light Ultraviolet illumination UV illuminations Wide area Xenopus laevis oocytes Alcohols Calcium Confocal microscopy Enzyme activity Fluorescence Fluorescence microscopy Mercury (metal) Optical fibers Photolysis Sugars Ultraviolet radiation Microscopes Xenopus laevis 2 nitrophenyl EGTA 2-nitrophenyl-EGTA calcium drug derivative egtazic acid inositol 1,4,5 trisphosphate animal article calcium signaling chemistry confocal microscopy equipment design instrumentation metabolism methodology oocyte photolysis physiology statistical model ultraviolet radiation Xenopus laevis Animals Calcium Calcium Signaling Egtazic Acid Equipment Design Inositol 1,4,5-Trisphosphate Linear Models Microscopy, Confocal Oocytes Photolysis Ultraviolet Rays Xenopus laevis The flash photolysis of "caged" compounds is a powerful experimental technique for producing rapid changes in concentrations of bioactive signaling molecules. These caged compounds are inactive and become active when illuminated with ultraviolet light. This paper describes an inexpensive adaptation of an Olympus confocal microscope that uses as source of ultraviolet light the mercury lamp that comes with the microscope for conventional fluorescence microscopy. The ultraviolet illumination from the lamp (350 - 400 nm) enters through an optical fiber that is coupled to a nonconventional port of the microscope. The modification allows to perform the photolysis of caged compounds over wide areas (~200 μm) and obtain confocal fluorescence images simultaneously. By controlling the ultraviolet illumination exposure time and intensity it is possible to regulate the amount of photolyzed compounds. In the paper we characterize the properties of the system and show its capabilities with experiments done in aqueous solution and in Xenopus Laevis oocytes. The latter demonstrate its applicability for the study of Inositol 1,4,5-trisphosphate-mediated intracellular calcium signals. © 2011 Society of Photo-Optical Instrumentation Engineers (SPIE). 2011 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10833668_v16_n6_p_Sigaut http://hdl.handle.net/20.500.12110/paper_10833668_v16_n6_p_Sigaut
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Cage compound Calcium signal Confocal microscopy Inositol 1,4,5-trisphosphate receptor Photolysis UV illumination Cage compounds Confocal fluorescence Confocal microscopes Experimental techniques Exposure-time Flash photolysis Inositol 1,4,5-trisphosphate receptor Intracellular calcium Olympus Rapid changes Signaling molecules Ultra-violet light Ultraviolet illumination UV illuminations Wide area Xenopus laevis oocytes Alcohols Calcium Confocal microscopy Enzyme activity Fluorescence Fluorescence microscopy Mercury (metal) Optical fibers Photolysis Sugars Ultraviolet radiation Microscopes Xenopus laevis 2 nitrophenyl EGTA 2-nitrophenyl-EGTA calcium drug derivative egtazic acid inositol 1,4,5 trisphosphate animal article calcium signaling chemistry confocal microscopy equipment design instrumentation metabolism methodology oocyte photolysis physiology statistical model ultraviolet radiation Xenopus laevis Animals Calcium Calcium Signaling Egtazic Acid Equipment Design Inositol 1,4,5-Trisphosphate Linear Models Microscopy, Confocal Oocytes Photolysis Ultraviolet Rays Xenopus laevis
spellingShingle	Cage compound Calcium signal Confocal microscopy Inositol 1,4,5-trisphosphate receptor Photolysis UV illumination Cage compounds Confocal fluorescence Confocal microscopes Experimental techniques Exposure-time Flash photolysis Inositol 1,4,5-trisphosphate receptor Intracellular calcium Olympus Rapid changes Signaling molecules Ultra-violet light Ultraviolet illumination UV illuminations Wide area Xenopus laevis oocytes Alcohols Calcium Confocal microscopy Enzyme activity Fluorescence Fluorescence microscopy Mercury (metal) Optical fibers Photolysis Sugars Ultraviolet radiation Microscopes Xenopus laevis 2 nitrophenyl EGTA 2-nitrophenyl-EGTA calcium drug derivative egtazic acid inositol 1,4,5 trisphosphate animal article calcium signaling chemistry confocal microscopy equipment design instrumentation metabolism methodology oocyte photolysis physiology statistical model ultraviolet radiation Xenopus laevis Animals Calcium Calcium Signaling Egtazic Acid Equipment Design Inositol 1,4,5-Trisphosphate Linear Models Microscopy, Confocal Oocytes Photolysis Ultraviolet Rays Xenopus laevis Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
topic_facet	Cage compound Calcium signal Confocal microscopy Inositol 1,4,5-trisphosphate receptor Photolysis UV illumination Cage compounds Confocal fluorescence Confocal microscopes Experimental techniques Exposure-time Flash photolysis Inositol 1,4,5-trisphosphate receptor Intracellular calcium Olympus Rapid changes Signaling molecules Ultra-violet light Ultraviolet illumination UV illuminations Wide area Xenopus laevis oocytes Alcohols Calcium Confocal microscopy Enzyme activity Fluorescence Fluorescence microscopy Mercury (metal) Optical fibers Photolysis Sugars Ultraviolet radiation Microscopes Xenopus laevis 2 nitrophenyl EGTA 2-nitrophenyl-EGTA calcium drug derivative egtazic acid inositol 1,4,5 trisphosphate animal article calcium signaling chemistry confocal microscopy equipment design instrumentation metabolism methodology oocyte photolysis physiology statistical model ultraviolet radiation Xenopus laevis Animals Calcium Calcium Signaling Egtazic Acid Equipment Design Inositol 1,4,5-Trisphosphate Linear Models Microscopy, Confocal Oocytes Photolysis Ultraviolet Rays Xenopus laevis
description	The flash photolysis of "caged" compounds is a powerful experimental technique for producing rapid changes in concentrations of bioactive signaling molecules. These caged compounds are inactive and become active when illuminated with ultraviolet light. This paper describes an inexpensive adaptation of an Olympus confocal microscope that uses as source of ultraviolet light the mercury lamp that comes with the microscope for conventional fluorescence microscopy. The ultraviolet illumination from the lamp (350 - 400 nm) enters through an optical fiber that is coupled to a nonconventional port of the microscope. The modification allows to perform the photolysis of caged compounds over wide areas (~200 μm) and obtain confocal fluorescence images simultaneously. By controlling the ultraviolet illumination exposure time and intensity it is possible to regulate the amount of photolyzed compounds. In the paper we characterize the properties of the system and show its capabilities with experiments done in aqueous solution and in Xenopus Laevis oocytes. The latter demonstrate its applicability for the study of Inositol 1,4,5-trisphosphate-mediated intracellular calcium signals. © 2011 Society of Photo-Optical Instrumentation Engineers (SPIE).
title	Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
title_short	Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
title_full	Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
title_fullStr	Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
title_full_unstemmed	Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals
title_sort	custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of inositol 1,4,5-trisphosphate-mediated calcium signals
publishDate	2011
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10833668_v16_n6_p_Sigaut http://hdl.handle.net/20.500.12110/paper_10833668_v16_n6_p_Sigaut
_version_	1768543909645910016

Custom-made modification of a commercial confocal microscope to photolyze caged compounds using the conventional illumination module and its application to the observation of Inositol 1,4,5-trisphosphate-mediated calcium signals

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