RNAssist

Cell & tissue fixation

In addition to its proven biomolecule stabilisation properties, vivoPHIX is an exceptionally effective universal cell and tissue fixative that has been successfully used for samples as diverse as plants, parasites, invertebrates and mammals. All tested mammalian tissues types have been fixed with vivoPHIX including fat rich tissues such as adipose and brain, protein rich such as skeletal and cardiac muscle, and standard tissues types including pancreas, liver and kidney. Fixed tissues can be used for standard paraffin embedding, sectioning and staining for H&E, IHC, IF, RNAscope, GFP imaging and FISH.

Multifunctional Staining

Diverse detection case study; Various vivoPHIX fixed sample types were imaged using IHC (anti-SMA with mouse kidney), IF (anti-ki67 with mouse stomach), ISH (RNAscope with mouse stomach), IF (Trypanosomes), RFP (Plasmodium), FISH (nascent transcripts in mouse stem cell nuclei), IHC (anti-CD20 with Zika virus infected spleen), IF (anti-FP and DAPI staining with mouse skin), crystal violet staining (human pancreatic adenocarcinoma). Thanks to Irina Pshenichnaya; Maud Borensztein; Roberto Bandiera; Martyna Popis, Uni. Cambridge; Giuseppe Diaferia, Istituto Europeo di Oncologia; Derrick Robinson, Uni. Bordeaux; Virginia Howick, Sanger Institute; Debbie Ferguson, NIBSC.

Multifunctional Staining

(1) Immunohistochemistry (IHC)

Simple mouse (i) brain, (ii) kidney and (iii) intestine fixation with vivoPHIX followed by standard paraffin embedding, sections were stained with anti-Smooth Muscle Actin (SMA).

(2) Immunohistochemistry (IHC)

Simple mouse (i) & (ii) kidney, and (iii) brain fixation with vivoPHIX followed by standard paraffin embedding, sections were used for IHC and stained with anti-b-catenin.

(3) Immunohistochemistry (IHC)

Simple (i) lung, (ii) kidney or (iii) spleen fixation with vivoPHIX followed by standard paraffin embedding, after epitope retrieval with HIER1 for 30 minutes, sections were used for IHC and stained with anti-CD20. Thanks to Dr. D. Ferguson, NIBSC, UK.

Immunofluorescence (IF)

vivoPHIX fixed mouse stomach, paraffin embedded and stained with anti-ki67 (nuclear) antibody and a fluorescent secondary antibody. Thanks to Dr. I. Pshenichnaya, University of Cambridge, UK.

in situ hybridization (ISH)

Comparison of in situ hybridisation signals using either NBF or vivoPHIX fixed mouse liver, paraffin embedding and hybridisation/staining with an oligodT probe (Ventana). Non-specific binding of the probe can be seen with the NBF but not with the vivoPHIX fixed sample.

Fluorescent protein (RFP & GFP)

vivoPHIX fixed fluorescent proteins expressed in (i) Plasmodium (RFP), and (ii) Drosophila larvae (GFP), maintain their fluorescence for several hours in vivoPHIX. Thanks to G. Howick and M. Lawniczak, Sanger Institute, UK, and  D. Xu, University of Oxford.

vivoPHIX Purification Protocol

Option A or B

rnassist-purification-protocol

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