NDLI: Axial super resolution topography of focal adhesion by confocal microscopy.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Chiu, Chi-Li Gratton, Enrico
Description	Author Affiliation: Chiu CL ( Department of Developmental and Cell Biology, University of California, Irvine, California)
Abstract	The protein organization within focal adhesions has been studied by state-of-the-art super resolution methods because of its thin structure, well below diffraction limit. However, to achieve high axial resolution, most of the current approaches rely on either sophisticated optics or diligent sample preparation, limiting their application. In this report we present a phasor-based method that can be applied to fluorescent samples to determine the precise axial position of proteins using a conventional confocal microscope. We demonstrate that with about 4,000 photon counts collected along a z-scan, axial localization precision close to 10 nm is achievable. We show that, with within 10 nm, the axial location of paxillin, FAK, and talin is similar at focal adhesion sites, while F-actin shows a sharp increase in height towards the cell center. We further demonstrated the live imaging capability of this method. With the advantage of simple data acquisition and no special instrument requirement, this approach could have wide dissemination and application potentials.
File Format	HTM / HTML
ISSN	1059910X
e-ISSN	10970029
DOI	10.1002/jemt.22267
Journal	Microscopy Research and Technique
Issue Number	10
Volume Number	76
Language	English
Publisher	Wiley-Liss
Publisher Date	2013-10-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Chemistry Techniques Focal Adhesions Chemistry Ultrastructure Microscopy, Confocal Proteins Animals Cell Line Cricetinae Research Support, N.i.h., Extramural
Content Type	Text
Resource Type	Article
Subject	Anatomy Histology Instrumentation Medical Laboratory Technology

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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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