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10.1101/pdb.prot067926 http://scihub22266oqcxt.onion/10.1101/pdb.prot067926 C4782786!4782786 !22301661     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\22301661 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Cold+Spring+Harb+Protoc 2012 ; 2012 (2 ): 242-5 Nephropedia Template TP {{tp|p=22301661 |t=2012. Single-molecule high-resolution colocalization of single probes |pdf=|usr=}}{{22301661 }} gab.com Text Single-molecule high-resolution colocalization of single probes JO: Cold Spring Harb Protoc http://www.kidney.de/pget.php?&tw=1&pmid=22301661 #FOAvip Colocalization of fluorescent probes is commonly used in cell biology to discern the proximity of two proteins in the cell. Considering that the resolution limit of optical microscopy is on the order of 250 nm, there has not been a need for high-resolution colocalization techniques. However, with the advent of higher resolution techniques for cell biology and single-molecule biophysics, colocalization must also improve. For diffraction-limited applications, a geometric transformation (i.e., translation, scaling, and rotation) is typically applied to one color channel to align it with the other; however, to achieve high-resolution colocalization, this is not sufficient. Single-molecule high-resolution colocalization (SHREC) of single probes uses the local weighted mean transformation to achieve a colocalization resolution of at least 10 nm. This protocol describes the acquisition of registration data and the analysis required to obtain a high-resolution mapping between imaging channels. The total internal reflection fluorescence microscope (TIRFM) system described is designed to excite and image the fluorescent probes Cy3 and Cy5. Modifications may be required depending on the requirements of the individual study. Twit Text FOAVip Single-molecule high-resolution colocalization of single probes ????Cold Spring Harb Protoc http://www.kidney.de/pget.php?&tw=1&pmid=22301661 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=22301661 #FOAvip English Wikipedia <ref>{{Cite pmid|22301661 }}</ref> Single-molecule high-resolution colocalization of single probes #MMPMID22301661 Churchman LS ; Spudich JA Cold Spring Harb Protoc 2012[Feb]; 2012 (2 ): 242-5 PMID22301661 show ga Colocalization of fluorescent probes is commonly used in cell biology to discern the proximity of two proteins in the cell. Considering that the resolution limit of optical microscopy is on the order of 250 nm, there has not been a need for high-resolution colocalization techniques. However, with the advent of higher resolution techniques for cell biology and single-molecule biophysics, colocalization must also improve. For diffraction-limited applications, a geometric transformation (i.e., translation, scaling, and rotation) is typically applied to one color channel to align it with the other; however, to achieve high-resolution colocalization, this is not sufficient. Single-molecule high-resolution colocalization (SHREC) of single probes uses the local weighted mean transformation to achieve a colocalization resolution of at least 10 nm. This protocol describes the acquisition of registration data and the analysis required to obtain a high-resolution mapping between imaging channels. The total internal reflection fluorescence microscope (TIRFM) system described is designed to excite and image the fluorescent probes Cy3 and Cy5. Modifications may be required depending on the requirements of the individual study. |Cytological Techniques/*methods [MESH] |Fluorescent Dyes/*analysis [MESH] |Image Processing, Computer-Assisted/*methods [MESH] |Microscopy, Confocal/*methods [MESH] |Microscopy, Fluorescence/methods [MESH]Single-molecule high-resolution colocalization of single probes (epmc).pdf DeepDyve Pubget Overpricing