Research Histology Services

Immunohistochemistry and Slide Scanning

What is Immunohistochemistry? This is a process by which specific antigens (i.e. proteins) are detected in cells of a tissue by utilizing the principles of the immune system whereby antibodies are generated and are specifically designed to target a particular antigen. These antibody-antigen interactions can then be visualized by a variety of means. For more information please check out our Instructional Video.

What is the difference between Chromogen and Immunofluorescent Detection?

  • Chromogen based detection uses secondary antibodies linked to an enzyme that can convert a substrate into a colored precipitate. This precipitate is then deposited on the tissue and can be detected using standand light microscopy. This method can be limited if the customer wants to evaluate more than one antigen at a time. A variety of color choices are available.
  • Immunofluorescent based detection uses secondary antibodies linked to fluorophores that which can be visualized when excited by a laser or UV light. This method of detection can result in increased sensitivity and works well when wanting to multiplex (i.e. visualize multiple antigens) at one time. A variety of color choices are available. This method can be limiting if the customer does not have access to the proper type of microscope.
  • If you are unsure which method may be best for your particular needs, please contact RHS for a consultation.

Can you recommend an antibody for my needs? RHS is happy to help you choose a new antibody from commercials vendors. Although this does not guarantee that the antibody will work, RHS offers the Antibody Work-up Service to test new antibodies. We encourage you to see if a commercial vendor offers trial sizes or has a program for testing antibodies in unpublished uses. For a list of antibodies which we have successfully used for customers in the past, please check our list. Antibodies chosen from this list may not require an Antibody Work-up fee if your experiment is in the same tissue used by our previous customer.

Do you have more than 10 slides for immunohistochemistry? RHS has a ThermoScientific Lab Vision Autostainer 360 with PT Module for automation of immunohistochemical stains. As such, if your experiment has a large number of slides, we ask that you provide enough antibody so that we can utilize the autostainer. The autostainer cuts down on variability from one batch of staining to another and can help reduce employee costs. This will cost the customer more money for primary antibody but saves customers money in the long run by reducing charges for employee time. An RHS member can help you determine how much antibody you will need to provide for your experiment.

Antibody Work-up for Immunohistochemistry

  • RHS offers this service to determine the optimum conditions for use of a new antibody for use in immunohistochemical procedures on either paraffin or frozen sections. This is a required service for any new antibody provided to RHS unless the customer provides a protocol. This service does not guarantee a working antibody.
  • What is included in this service? Antibodies are tested on control tissue using various antigen retrieval methods to determine the optimal staining conditions. Antibodies are then titrated to determine the best concentration for staining using control tissues. RHS provides most control tissues for human or mouse tissues. For rare tissues, other species, or special transgenic reporter mice, RHS may request the customer to provide their own control tissue. The customer is required to provide primary antibody.
  • What is a typical work up for paraffin sections? Antibody tested at manufacturer’s recommended dilution (or 1:100 if not specified). Three methods of Heat-Induced Antigen Retrieval using a steamer for 30 minutes on control tissue.
    • Target Retrieval Solution (Dako, S1699; modified sodium citrate buffer, pH 6.1)
    • Target Retrieval Solution, pH 9 (Dako, S2367; Tris/EDTA buffer, pH 9)
    • Lab Vision™ EDTA Buffer for Heat-Induced Epitope Retrieval (ThermoScientific, AP-9004-125; 10mM EDTA buffer, pH 8.0)
    • Negative Controls without Primary Antibody
    • All slides are developed using chromogen based IHC with AEC.
  • What if the typical work-up is not perfect or does not work well? RHS is happy to continue to optimize the antibody conditions with customer approval. Each attempt to get a customers antibody working will incur an additional work-up fee. Representative conditions that could be tried:
    • Various dilutions of antibody
    • Alternate antigen retrieval methods
    • Other control tissues
    • Recommended protocols in published papers (that differ from manufacturer’s recommendations)
  • How are work-ups in frozen sections different? If staining is desired on frozen sections, enzymatic methods of antigen retrieval are tested rather than heat-induced antigen retrieval methods.
  • Once antibody is working to the satisfaction of the customer and core director, the customer's experimental samples will be stained and charged according to pricing below.



Chromogen Based Immunohistochemistry


Immunofluorescence Based Immunohistochemistry


Multiplex Labelling with Qdot Nanocrystals (Fluorscence Based IHC)


Slide Scanning

RHS offers whole slide scanning services. Slides that have already been stained (special stains or IHC, chromogen or fluorescent) can be digitized for analysis as a service. We utilize Zeiss Mirax and Axioscan technologies for this service.  Analysis is only done on a collaboration basis but RHS is happy to discuss your analysis needs and potential costs associated with such an analysis. Please contact Drew Lesniak or Michelle Wood-Trageser for more information regarding such services.

Brightfield Slide Scanning


Fluorescence Slide Scanning