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.
**Substrate dissolves in acetone wash.
***A slight color change in ImmPACT NovaRED and Vector NovaRED reaction produt may be seen using methyl green.


Resources

Immunohistochemistry Resource Guide

The Immunohistochemistry Resource Guide will help you navigate through the many choices of detection systems, substrates, and ancillary reagents that complete your workflow.
Download

IHC Troubleshooting Guide for ABC Systems

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.

Download

IHC Troubleshooting Guide for Polymer Kits

A quick reference guide that outlines the systematic controls to help identify background sources in an immunohistochemistry protocol polymer detection systems.  Also troubleshoot issues of weak or no staining. Laminated.

Download