I need to prepare Antifading (like Citifluor) solution for FISH technique.
Dear Graciana,
It exists different protocols to prepare Antifading solutions:
https://sites.google.com/site/singlemoleculernafish/reagents-and-equipment
https://books.google.ch/books?id=TMP4CCAu_MYC&pg=PA90&dq=Antifading++fish&hl=de&sa=X&ved=0ahUKEwi2lbD0hbzQAhUHPBQKHTKiCGYQ6AEIKTAA#v=onepage&q=Antifading%20%20fish&f=false
Markey FB, Ruezinsky W, Tyagi S, Batish M. Fusion FISH Imaging: Single-Molecule Detection of Gene Fusion Transcripts In Situ. Pack S, editor. PLoS ONE. Public Library of Science; 2014;9(3):e93488.
Semrau S, Crosetto N, Bienko M, Boni M, Bernasconi P, Chiarle R, et al. FuseFISH: Robust Detection of Transcribed Gene Fusions in Single Cells. CellReports. 2013 Dec 24.
Yang B, Treweek JB, Kulkarni RP, Deverman BE, Chen C-K, Lubeck E, et al. Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing. Cell. 2014 Jul 31
http://onlinelibrary.wiley.com/doi/10.1002/cyto.990190213/full
Maybe you will find something useful inside one of those. The first link gives a complete FISH Anti-fade Buffer composition, but I think that you need to double check with other publications to see if this is useful.
Hope it helped,
Regards,
Mathieu
Hi
We've always had great results in microscopy using the method from the Mitchison lab. I mix together 0.5 % p-phenylenediamine in 20 mM Tris, pH 8.8, 90 % glycerol. I do a large volume (say 500ml), transfer it to a cylinder and gently bubble nitrogen through it for 4 hours or so, keeping it dark.
I aliquot it into syringes, cap them and store them in the -80. I keep one at -20 for regular use. A single syringe will take months and months to dis-colour, so a single prep should keep you going for years!
I also recommend their 'AbDil' solution for antibody staining. You can see their recipes at http://mitchison.hms.harvard.edu/files/mitchisonlab/files/fluorescence_procedures_for_the_actin_and_tubulin_cytoskeleton_in_fixed_cells.pdf
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