CYO Stain

v DAB and Cytochrome C

o Dangerous Chemicals

o Gloves and mask when handling

o Wash instruments with bleach after use

o Left over medium requires bleach to be added and left until solution turns black

Purpose

Ø To observe metabolic activity in tissue

Materials and Procedure

Do not stir vigorously – this will add oxygen to the medium influencing the intensity of the stain. Be consistent with stirring speed intra/inter batch

CYO Staining Medium

In

· 19.2 ml of 0.1 M PB (pH 7.40)

Add

· 0.8 g Sucrose

·

Light Sensitive

Wrap flask with Al foil

· 10.0 mg Diaminobenzidine (DAB)

· 4.0 mg Cytochrome C Type III

Staining

1. Rinse Tissue 3x10 min in 0.1 M PB

2. Incubate in CYO stain for ~2 hrs on agitator

3. Rinse 3x10 min in 0.1 M PB

4. Mount on Pig Gel slides ® allow to dry ® cover slip