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10.3791/62067 http://scihub22266oqcxt.onion/10.3791/62067 33427241!ä!33427241 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Vis+Exp 2020 ; ä (166): ä Nephropedia Template TP {{tp|p=33427241|t=2020. Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization |pdf=|usr=}}{{33427241}} gab.com Text Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization JO: J Vis Exp http://www.kidney.de/pget.php?&tw=1&pmid=33427241 #FOAvip This manuscript provides a protocol for in situ hybridization chain reaction (HCR) coupled with immunofluorescence to visualize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in cell line and three-dimensional (3D) cultures of human airway epithelium. The method allows highly specific and sensitive visualization of viral RNA by relying on HCR initiated by probe localization. Split-initiator probes help amplify the signal by fluorescently labeled amplifiers, resulting in negligible background fluorescence in confocal microscopy. Labeling amplifiers with different fluorescent dyes facilitates the simultaneous recognition of various targets. This, in turn, allows the mapping of the infection in tissues to better understand viral pathogenesis and replication at the single-cell level. Coupling this method with immunofluorescence may facilitate better understanding of host-virus interactions, including alternation of the host epigenome and immune response pathways. Owing to sensitive and specific HCR technology, this protocol can also be used as a diagnostic tool. It is also important to remember that the technique may be modified easily to enable detection of any RNA, including non-coding RNAs and RNA viruses that may emerge in the future. Twit Text FOAVip Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization ????J Vis Exp http://www.kidney.de/pget.php?&tw=1&pmid=33427241 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33427241 #FOAvip English Wikipedia <ref>{{Cite pmid|33427241}}</ref> Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybridization #MMPMID33427241 Kula-Pacurar A; Wadas J; Suder A; Szczepanski A; Milewska A; Ochman M; Stacel T; Pyrc K J Vis Exp 2020[Dec]; ä (166): ä PMID33427241show ga This manuscript provides a protocol for in situ hybridization chain reaction (HCR) coupled with immunofluorescence to visualize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in cell line and three-dimensional (3D) cultures of human airway epithelium. The method allows highly specific and sensitive visualization of viral RNA by relying on HCR initiated by probe localization. Split-initiator probes help amplify the signal by fluorescently labeled amplifiers, resulting in negligible background fluorescence in confocal microscopy. Labeling amplifiers with different fluorescent dyes facilitates the simultaneous recognition of various targets. This, in turn, allows the mapping of the infection in tissues to better understand viral pathogenesis and replication at the single-cell level. Coupling this method with immunofluorescence may facilitate better understanding of host-virus interactions, including alternation of the host epigenome and immune response pathways. Owing to sensitive and specific HCR technology, this protocol can also be used as a diagnostic tool. It is also important to remember that the technique may be modified easily to enable detection of any RNA, including non-coding RNAs and RNA viruses that may emerge in the future. |*COVID-19 Testing[MESH] |*RNA, Viral[MESH] |Animals[MESH] |COVID-19/*diagnosis/*virology[MESH] |Chlorocebus aethiops[MESH] |Fluorescence[MESH] |Humans[MESH] |In Situ Hybridization, Fluorescence[MESH] |Respiratory Mucosa/*virology[MESH] |Respiratory System[MESH] |SARS-CoV-2/*genetics[MESH]Visualization of SARS-CoV-2 using Immuno RNA-Fluorescence In Situ Hybr(epmc).pdf DeepDyve Pubget Overpricing