IHC, ISH and ICC Resources

All of these acronyms can be confusing – what’s the difference between these techniques?

Immunohistochemistry (IHC) and immunocytochemistry (ICC) are techniques employed to localise antigen expression and are dependent on specific epitope-antibody interactions. IHC refers to the use of tissue sections, whereas ICC describes the use of cultured cells or cell suspensions. In both methods, positive staining is visualised using a molecular label, which can be fluorescent or chromogenic. Briefly, samples are fixed to preserve cellular integrity, and then subjected to incubation with blocking reagents to prevent non-specific binding of the antibodies. Samples are subsequently incubated with primary and secondary antibodies, and the signal is visualised for microscopic analysis.

Technically, IHC/ICC are relatively simple and straightforward experimental methods. However, there are many variables which must be recognised and optimised for each individual IHC/ICC study. The most challenging aspect of these techniques is determining the experimental conditions necessary to generate a strong and specific signal for each antigen of interest. For example, detection of an abundant protein in cultured cells may require a short fixation period, minimal blocking, and may be compatible with direct visualisation using a fluorochrome-conjugated primary antibody. In contrast, detection of a phosphorylation-dependent epitope in a section of frozen tissue may require antigen retrieval and be dependent on amplified chromogenic visualisation. 

The following protocols and troubleshooting guides developed by Bio-Techne sister companies R&D Systems and Novus Biologicals can help guide you through the process of successful design and optimisation of your IHC and ICC protocols.

IHC / ICC Protocol Guide
Download R&D Systems^® IHC/ICC Protocol Guide to get protocols, techniques, and troubleshooting suggestions to help you design and fine-tune your IHC/ICC experiment.

ICC Handbook
This Immunocytochemistry (ICC) Handbook offers an introduction to ICC with a focus on the conceptual basics of multicolor ICC/IF, including detailed protocols and troubleshooting tips.

IHC Handbook
Download this Immunohistochemistry (IHC) Handbook for a clear and concise introduction to IHC for researchers new to the technique or lab scientists in search of a quick refresher.

Insights on Dual ISH-IHC 
Download and read interviews about how four researchers used dual ISH-IHC assays in their labs.

The Founder of ACD Bio Dr Yuling Luo sat down with us and answered our top 20 FAQs on In Situ Hybridization.

Dr Yuling Luo, President of Advanced Cell Diagnostics presents a seminar on Next Generation RNA In Situ Hybridization Technologies For Research and Diagnostics.

RNA Scope Reference Guide

The Definitive ISH Guide
Guidelines and protocols for obtaining optimal RNAscope^® in situ hybridization results with any tissue type from any species

Additional Resources

Read a recent article by Novus Biologicals – Frontiers in immunohistochemical analysis

 Making IHC Problem Free 

R&D Systems offers four secondary antibodies that will transform IHC. These Anti-IgG VisUCyte^™ HRP Polymer-Conjugated Antibodies are biotin-free detection reagents that overcome many problems commonly associated with the avidin-biotin detection system. Learn more.

Free Webinar!

Need to validate cell types with antibodies from a single host source? This webinar details a novel technique we developed for triple-labeling immunofluorescence immunohistochemistry (IHC) that can be used with primary antibodies from the same source. Watch now!

Mona Lisa IHC/ICC Mosaic Wall Poster

Turn your lab into an art gallery with our new Mona Lisa mosaic wall poster. Thous ands of IHC and ICC images generated with our antibodies were used to create this spectacular wall poster.

Email lifescience@invitro.com.au to request your copy today!