Counterstains and Special Stains
Histology stains for improved target visualization and localization
Histology stains and counterstains introduce color to specific cellular structures to provide contrast to colored enzyme substrates. This makes targets easier to see, which makes it easier to interpret tissue morphology and cell structure.
Counterstains
Counterstains stain cell nuclei and are packaged as convenient, ready-to-use solutions for use on slides or in staining dishes. Available to you (see selection guide below):
- Vector® Hematoxylin — based on Gill’s formulation, can be adjusted for optimal intensity; suitable for use with non-aqueous and aqueous mounting media and is mercury- and alcohol-free
- Vector® Hematoxylin QS — based on a modification of Mayer’s hematoxylin and was developed specifically for immunocytochemistry (ICC). It stains within 45 sec, is suitable for use with non-aqueous and aqueous mounting media, and is mercury-free
- Vector® Methyl Green — offers a simple, two-step procedure and is an excellent alternative in multiple antigen labeling when hematoxylin obscure substrate color. It is suitable for use with non-aqueous mounting media
- Vector® Nuclear Fast Red — offers a fast, one-step protocol and is an excellent alternative in multiple antigen labeling when hematoxylin obscure substrate color
- Hematoxylin and Eosin Stain Kit — includes a modified Eosin for cytoplasm staining and Hematoxylin for crisp, intense blue nuclei
Special Stains
Use the Alcian Blue (pH 2.5) Stain Kit to visualize sulfated and carboxylated acid mucopolysaccharides and sulfated and carboxylated sialomucins (glycoproteins).
- Acidic sulfated Mucosubstances (blue)
- Hyaluronic acid: Blue
- Sialomucins (blue)
- Nuclei (red)
Counterstain / Substrate Compatibility
This table is designed as a reference to determine the optimal counterstain / substrate combination for your application. Consideration should be given to tissue type, antigen unmasking protocol, and other detection parameters to achieve the desired staining intensity.
Substrate | Catalog Number | Hematoxylin (H-3404) Hematoxylin QS (H-3401) |
Methyl Green (H-3402) | Nuclear Fast Red (H-3404) |
ImmPACT DAB (brown) |
SK-4105 | Excellent Contrast | Excellent Contrast | Fair Contrast |
ImmPACT DAB EqV (brown) |
SK-4103 | Excellent Contrast | Excellent Contrast | Fair Contrast |
DAB (brown) |
SK-4100 | Excellent Contrast | Excellent Contrast | Fair Contrast |
DAB-Ni (gray-black) |
SK-4100 | Excellent Contrast | Fair Contrast* | Good Contrast |
ImmPACT AEC (red) |
SK-4205 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
ImmPACT AMEC Red (red) |
SK-4285 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
AEC (red) |
SK-4200 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
TMB (blue) |
SK-4400 | Color Incompatibility | Counterstain Incompatibility | Excellent Contrast |
ImmPACT VIP (purple) |
SK-4605 | Fair Contrast | Excellent Contrast | Poor Contrast |
Vector VIP (purple) |
SK-4600 | Fair Contrast | Excellent Contrast | Poor Contrast |
ImmPACT SG (blue-gray) |
SK-4705 | Poor Contrast | Good Contrast | Excellent Contrast |
SG (blue-gray) |
SK-4700 | Poor Contrast | Good Contrast | Excellent Contrast |
ImmPACT NovaRED (red) |
SK-4805 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
Vector NovaRED (red) |
SK-4800 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
ImmPACT Vector Red (magenta) |
SK-5105 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
Vector Red (magenta) |
SK-5100 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
Vector Black (black) |
SK-5200 | Excellent Contrast | Excellent Contrast* | Excellent Contrast |
Vector Blue (blue) |
SK-5300 | Color Incompatibility | Good Contrast | Excellent Contrast |
BCIP / NBT (indigo) |
SK-5400 | Color Incompatibility | Excellent Contrast* | Excellent Contrast |
*This substrate shows a slight decrease in sensitivity following the methyl green protocol. This decrease can be minimized by reducing the heat incubation and acetone rinse times. |
Resources
A quick reference guide that outlines the systematic controls to help identify background sources in an immunohistochemistry protocol using ABC systems. Also troubleshoot issues of weak or no staining. Laminated.