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High-performance probes for light and electron microscopy.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Viswanathan, Sarada Williams, Megan E. Bloss, Erik B. Stasevich, Timothy J. Speer, Colenso M. Nern, Aljoscha Pfeiffer, Barret D. Hooks, Bryan M. Li, Wei-Ping English, Brian P. Tian, Teresa Henry, Gilbert L. Macklin, John J. Patel, Ronak Gerfen, Charles R. Zhuang, Xiaowei Wang, Yalin Rubin, Gerald M. Looger, Loren L.
Description	Country affiliation: United States Author Affiliation: Viswanathan S ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Williams ME ( Department of Neurobiology &Anatomy, University of Utah, Salt Lake City, Utah, USA.); Bloss EB ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Stasevich TJ ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Speer CM ( Howard Hughes Medical Institute, Department of Chemistry &Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.); Nern A ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Pfeiffer BD ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Hooks BM ( 1] Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA. [2] Laboratory of Systems Neuroscience, National Institute of Mental Health, Bethesda, Maryland, USA.); Li WP ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); English BP ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Tian T ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Henry GL ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Macklin JJ ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Patel R ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Gerfen CR ( Laboratory of Systems Neuroscience, National Institute of Mental Health, Bethesda, Maryland, USA.); Zhuang X ( 1] Howard Hughes Medical Institute, Department of Chemistry &Chemical Biology, Harvard University, Cambridge, Massachusetts, USA. [2] Howard Hughes Medical Institute, Department of Physics, Harvard University, Cambridge, Massachusetts, USA.); Wang Y ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Rubin GM ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.); Looger LL ( Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, USA.)
Abstract	We describe an engineered family of highly antigenic molecules based on GFP-like fluorescent proteins. These molecules contain numerous copies of peptide epitopes and simultaneously bind IgG antibodies at each location. These 'spaghetti monster' fluorescent proteins (smFPs) distributed well in neurons, notably into small dendrites, spines and axons. smFP immunolabeling localized weakly expressed proteins not well resolved with traditional epitope tags. By varying epitope and scaffold, we generated a diverse family of mutually orthogonal antigens. In cultured neurons and mouse and fly brains, smFP probes allowed robust, orthogonal multicolor visualization of proteins, cell populations and neuropil. smFP variants complement existing tracers and greatly increase the number of simultaneous imaging channels, and they performed well in advanced preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improved single-molecule image tracking and increased yield for RNA-seq. These probes facilitate new experiments in connectomics, transcriptomics and protein localization.
File Format	HTM / HTML
ISSN	15487091
e-ISSN	15487105
DOI	10.1038/nmeth.3365
Journal	Nature Methods
Issue Number	6
Volume Number	12
Language	English
Publisher	Nature Publishing Group
Publisher Date	2015-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Clinical Laboratory Techniques Luminescent Proteins Chemistry Microscopy, Electron Microscopy, Fluorescence Animals Antigens Brain Mapping Drosophila Mice Models, Molecular Molecular Sequence Data Neurons Protein Conformation Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology Biotechnology

Sl.	Authority	Responsibilities	Communication Details
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
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

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