1 [micro]g/ml tubulin-Alex Fluor 488 (Invitrogen, USA) for 30 min, Mito Tracker Red CMXRos (Invitrogen, USA) for 20 min, and 4',6-diamidino-2-phenylindole
Staining with 4',6-diamidino-2-phenylindole
showed that cells of CBS 10496 were monokaryotic (Figure 1, panel B).
For semi-quantitative assessment of apoptosis, cells were stained with 4',6-diamidino-2-phenylindole
(DAPI), which detects the DNA in them.
To examine the effect of AF on visualization of IF staining with CLSM, renal AF spectra were compared to spectra of 3 representative fluorophores: 4',6-diamidino-2-phenylindole
(DAPI), FITC, and Texas Red.
Curcumin (with over 80% curcumin and over 94% curcuminoid content purity), cis-platin (DDP), 3-[4,5-dimethylthiazol-2-y1]2,5-cliphenyltetrazolium bromide (MU), 4',6-diamidino-2-phenylindole
(DAPI) and zVAD-fmk were purchased from Sigma-Aldrich (St.
We purchased RPMI 1640 cell culture media, fetal bovine serum albumin, penicillin-streptomycin, 1X PBS, goat anti-mouse IgG Alexafluor 488, and 4',6-diamidino-2-phenylindole
(DAPI) from Invitrogen; recombinant human E (rhE)-selectin and P-selectin fused to IgG from R&D Systems; mouse anti-human prostate-specific membrane antigen (PSMA) monoclonal IgG from Abcam; protein-G from EMD Biosciences; and blotting-grade blocker nonfat dry milk from Bio-Rad Laboratories.
Fluorescent dyes are available that can be taken up by living cells to display nuclear morphology (eg, 4',6-diamidino-2-phenylindole
This system is based on positive selection for CTCs with antibodies to EpCAM and the staining of EpCAM-positive cells for CKs with 4',6-diamidino-2-phenylindole
and for CD45.
Nuclei were identified by 4',6-diamidino-2-phenylindole
dihydrochloride (DAPI, 1:10,000; Invitrogen).
ONL and INL thickness was measured on 4',6-diamidino-2-phenylindole
(DAPI)-stained slides using a calibrated filar micrometer eyepiece.
Nucleic acid was stained with 4',6-diamidino-2-phenylindole
and epithelial cells were stained with anti--cytokeratin-diamidino-2-phenylindole.
Detection approaches are based on (a) imaging (immunocytochemistry and immunofluorescence) through the use of specific markers for CTCs such as CKs (mainly CK-8, -18, and -19), leukocytes such as CD45, and cell viability via 4',6-diamidino-2-phenylindole
dihydrochloride (DAPI) staining--mainly by the FDA-approved CellSearch system; (b) molecular methods based on gene expression of specific markers such as CK-19; and (c) methods based on the detection of proteins secreted by immobilized CTCs, such as the EPISPOT assay.