High-performance probes for light and electron microscopy

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) distribute well in neuron...

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Main Authors:	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.
Format:	Online
Language:	English
Published:	2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4573404/

id	pubmed-4573404
recordtype	oai_dc
spelling	pubmed-45734042015-12-01 High-performance probes for light and electron microscopy 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. Article 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) distribute well in neurons, notably into small dendrites, spines and axons. smFP immunolabeling localizes 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 allow robust, orthogonal multi-color visualization of proteins, cell populations and neuropil. smFP variants complement existing tracers, greatly increase the number of simultaneous imaging channels, and perform well in advanced preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improve single-molecule image tracking and increase yield for RNA-Seq. These probes facilitate new experiments in connectomics, transcriptomics and protein localization. 2015-04-27 2015-06 /pmc/articles/PMC4573404/ /pubmed/25915120 http://dx.doi.org/10.1038/nmeth.3365 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
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.
spellingShingle	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. High-performance probes for light and electron microscopy
author_facet	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.
author_sort	Viswanathan, Sarada
title	High-performance probes for light and electron microscopy
title_short	High-performance probes for light and electron microscopy
title_full	High-performance probes for light and electron microscopy
title_fullStr	High-performance probes for light and electron microscopy
title_full_unstemmed	High-performance probes for light and electron microscopy
title_sort	high-performance probes for light and electron microscopy
description	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) distribute well in neurons, notably into small dendrites, spines and axons. smFP immunolabeling localizes 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 allow robust, orthogonal multi-color visualization of proteins, cell populations and neuropil. smFP variants complement existing tracers, greatly increase the number of simultaneous imaging channels, and perform well in advanced preparations such as array tomography, super-resolution fluorescence imaging and electron microscopy. In living cells, the probes improve single-molecule image tracking and increase yield for RNA-Seq. These probes facilitate new experiments in connectomics, transcriptomics and protein localization.
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4573404/
_version_	1613476689669521408

High-performance probes for light and electron microscopy

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