Fig. 2
From: Active centromere and chromosome identification in fixed cell lines
Workflow combining and comparing mFISH, cenFISH and CENP-IF. a Standard mFISH as suggested by MetaSystems. b cenFISH-mFISH: pan-centromeric FISH with pTRA7 α-satellite probes, designated as cenFISH, imaged with metaphase spreads coordinates recorded and underwent mFISH. c Freshly harvested cells swelled in hypotonic solution were cytospun onto microscope slide before immunofluorescence was performed using KCM buffer, cross-linked with 4 % formaldehyde, fixed and denatured with methanol-acetic acid, aged, underwent cenFISH and subsequently mFISH. d Immunofluorescence using TEEN and KB buffer was performed on cells stored in methanol-acetic acid fixative, cross-linked with 4 % formaldehyde before cenFISH and then mFISH was performed. Both methods c and d are CENP-IF-cenFISH-mFISH