Hello,

I was wondering whether anyone have encountered issues with Hoechst nuclei staining in 40um vibratome coronal brain sections of embryonic brain treated with HCl to reveal BrdU.

I am treating sections with 1M HCl for 2h at RT (rocker) followed by BrdU antibody incubation ON. After 2ary antibody incubation, sections are incubated with Hoechst for 10min at RT (rocker) followed by PBS washing (3 x 10min). Afterwards, the sections are mounted using Flouoromont G media.

Does anyone have any suggestion to solve the nuclei staining issue?

Thanks in advance.