NDLI: Pulsed local-field fluorescence microscopy: a new approach for measuring cellular signals in the beating heart

Content Provider	Springer Nature Link
Author	Mejía Alvarez, Rafael Man, Carlo Villalba Galea, Carlos A. del Valle Fernández, Luz Costa, Roberta Fill, Michael Gharbi, Tijani Escobar, Ariel L.
Copyright Year	2003
Abstract	In cardiac research, single-cell experimental models have been extensively used to study the molecular mechanisms of intracellular Ca2+ homeostasis. The results of these studies are usually extrapolated to the tissue level assuming that the phenomena studied at the cellular level are either similar in the intact organ, or only slightly modified by variables that exist at the whole-heart level. The validity of these assumptions has rarely been confirmed experimentally. Common obstacles associated with the study of intracellular Ca2+ signals in beating hearts include motion artifacts and spatio-temporal limitations of the recording system. In this work, action potentials and intracellular Ca2+ signals were measured in beating hearts from young rats, with spatio-temporal resolutions similar to cellular studies using a novel pulsed local-field fluorescence technique. This method was based on maximizing emitted fluorescence to increase the signal-to-noise ratio (S/N). The fluorescence emission of the indicator molecules was synchronized with brief (<1 ns), high-power (400 W) laser pulses, and the common mode noise of the fluorescence signal was differentially cancelled. To follow rapidly evolving signals, a highly sensitive and fast detection system was used (10 kHz). The spatial resolution was improved using a small (50–200 µm diameter) multimode fiberoptic. Mechanical artifacts were effectively reduced by inserting the fiberoptic into a "floating" glass micropipette sealed to the heart wall with negative pressure. Our results demonstrate that local-field fluorescence microscopy offers an outstanding experimental approach for studying physiological signals at the whole-organ level with the high spatio-temporal resolution common to normal cellular approaches.
Starting Page	747
Ending Page	758
Page Count	12
File Format	PDF
ISSN	00316768
Journal	Pflügers Archiv
Volume Number	445
Issue Number	6
e-ISSN	14322013
Language	English
Publisher	Springer-Verlag
Publisher Date	2003-01-14
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Physiology Physiology (medical) Clinical Biochemistry

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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