Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Nat+Methods 2015 ; 12 (6): 568-76 Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
High-performance probes for light and electron microscopy #MMPMID25915120
Viswanathan S; Williams ME; Bloss EB; Stasevich TJ; Speer CM; Nern A; Pfeiffer BD; Hooks BM; Li WP; English BP; Tian T; Henry GL; Macklin JJ; Patel R; Gerfen CR; Zhuang X; Wang Y; Rubin GM; Looger LL
Nat Methods 2015[Jun]; 12 (6): 568-76 PMID25915120show ga
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.