Hematoxylin & Eosin (H & E) Staining

Chemical Time
1 100%EtOH 1min.
2 100%EtOH 1min.
3 95% EtOH 1min.
4 95% EtOH 1min.
5 dH2O 1min.
6 dH2O 1min.
7 Hematoxylin 3min.
8 dH2O 30sec.
9 dH2O 30sec.
10 5% Glacial Acetic Acid 3 quick dips
11 dH2O 30 sec.
12 dH2O 30 sec.
13 Scott’s H 2O 15sec
14 dH 2O 1min.
15 dH 2O 1min.
16 Eosin 2min.
17 95% EtOH 30 sec.
18 95% EtOH 30 sec.
19 100%EtOH 30 sec.
20 100%EtOH 30 sec.
21 Xylene 1min.
22 Xylene 1min.
23 Permount  
Stain Color
Nuclei blue /black
Collagen and Skeletal Muscle bright red
Smooth Muscle pink / red (distinct from collagen)
RBC and Fibrin Bright Red/Orange