Nuclei were stained with 5 mg/ml 4',6-diamidino-2-phenylindole
(DAPI; Sigma) (Anway et al, 2003).
The oocytes were rinsed with distilled water and stained in 8 [micro]L of 4',6-diamidino-2-phenylindole
(DAPI) at a concentration of 2 [micro]g/mL (Barros e Silva & Guerra 2009).
The cells were then incubated with the primary antibodies and 4',6-diamidino-2-phenylindole
(DAPI) (Invitrogen, USA) overnight at 4[degrees]C and with secondary antibody at room temperature the next morning.
a-c) Values from three replicates with different superscripts denote a significant difference relative to other groups (p<0,05), TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; DAPI, 4',6-diamidino-2-phenylindole
Therefore, it is difficult or even impossible to manually relocate these sparse cells on the 4',6-diamidino-2-phenylindole
(DAPI) counterstain after hybridization, as morphologic details are lost.
Immunofluorescence was visualized using Alexa Fluor 488 and 594 antibodies (1:400, goat; Molecular Probes), 4',6-diamidino-2-phenylindole
(DAPI; Vector Labs), and HRP (1:400, goat; Jackson) + 3,3,-diaminobenzidine (DAB).
After 1 h staining at room temperature, the slides were washed and all cell nuclei were counter-stained with 4',6-diamidino-2-phenylindole
(DAPI (SouthernBiotech; Birmingham, United States)) and covered with ProLong[R] Gold Anti Fade.
Trifolin, propidium iodide (PI), and the 4',6-diamidino-2-phenylindole
(DAPI) stain were obtained from Sigma-Aldrich Co.
DNA was stained with 4',6-diamidino-2-phenylindole
dihydrochloride (DAPI; Sigma-Aldrich Co.
After washing with 4',6-diamidino-2-phenylindole
(DAPI) (Biochemicals), the cells were stained with DAPI, and observed by fluorescent confocal microscopy.
Nuclei were stained with 4',6-diamidino-2-phenylindole
(DAPI) at a dilution of 1:25,000 for 20 min at RT.
Following with sealing by using 4',6-diamidino-2-phenylindole
(DAPI) containing ProLong Gold Antifade Mountant (Life Technology), the concomitant nuclear staining was allowed in one procedure.