Bi-staining with Safranin-O and Alcian-blue stains – methods compared

Mixed bi-stain versus 2 single stains applied serially

Sunday 28/08/2018, 4 slides used for stain-test.
All 4 slides are 10ยต sections of Potato-stem in TS.

This test is to compare the results of bi-staining using Safranin and Alcian-blue stains. Stains used are, Safranin-O 1%(aq), Alcian-blue 1%(aq) acetic-acid.

Use of a 1:1 mixture of the two stains is compares to the 'traditional' use of the 2 stains separately, Safranin first, then Alcian-blue. All staining times were 5 minutes.

Slides 3 & 4:
Safranin/Alcian-blue bi-stain mixed, 5 minute stain then DIW rinse,

Slides 5 & 6:
Safranin for 5 minutes then DIW rinse followed by Alcian-blue for 5 minutes then DIW rinse.

All 4 slides were moved from DIW rins to 50%OH, 75%OH then 95%OH before being differentiated as needed using 1% acetic-acid OH, before being rinsed to OH then Histoclear for mounting.

Slide 3 at x10

Slide 3 at x25

Slide 4 at x10

Slide 4 at x25

Slide 5 at x10

Slide 5 at x25

Slide 6 at x10

Slide 6 at x25