destain

In another study conducted in 1995, comparing acridine orange and Auramine O as primary stains, the importance of methylene blue component in the destaining reagent has been highlighted, which provided reduced background fluorescence and better contrast making AFB easily visible (15).

Electrophoresis followed staining of the polyacrylamide gel for 20-30 minutes and destaining on a shaker until the disappearance of background color.

The gels were developed the next day using Coomassie Blue R-250 for 30 min and then destained using a destaining solution.

Under the dark, the gels were stained with Pro-Q Diamond phosphoprotein gel stain (Life Technologies, Carlsbad, CA) for 120 min at room temperature with gentle agitation and then washed three times with destaining solution [50 mM sodium acetate, pH 4.0 and 20% (v/v) acetonitrile] for 30 min.

After destaining, the MMP activity was detected as clear bands against the blue background.

Compared to classic AFB staining, Fite's acid-fast stain uses a milder destaining acid, as well as less alcohol during the staining process, which it suitable for more delicate mycobacterium, such as leprosy bacillus [12].

Alcian staining solution was removed and spheroids were washed with destaining solution (120 ml 98-100% ethanol + 80 ml acetic acid (98-100%)) 2x for 20 minutes.

The gel was stained with 0.1% Congo red dye for 30 min following destaining in 2M NaCl solution.

The staining procedure followed the manufacturer's recommendations, and all staining and destaining steps were protected from light.

It also permits preservation of cytological material by destaining and restaining with permanent stains.

After microscopic analysis of chromosome preparations by standard procedure (Giemsa staining), the identification of rearranged chromosomes was conducted by destaining and applying the G-band technique.