The terms immunohistochemistry, immunocytochemistry and immunofluorescence are often confused. When it comes to immunostaining these terms should not be used interchangeably regardless of the method.
The key differences are outlined below to ensure that anyone new to the field does not use the wrong term in an abstract or presentation. Firstly, consider these terms broken down into prefixes, roots and suffixes.
Root Word Meanings:
Immuno – an immunological technique (for example, the binding of antibodies to an antigen)
Histo – tissue (cells with the extracellular matrix)
Cyto – cells (cells without the extracellular matrix)
Chemistry – a method of detection using chemicals (for example, a change in color)
Fluorescence – using emission of light from an excited fluorophore for detection
Generally, the terms immunohistochemistry and immunocytochemistry are most often used correctly for their broad meaning to “use an antibody to detect an antigen” in a tissue or cell, respectively.
It is when people use a fluorophore for the detection of the primary antibody, be this through a direct conjugation to the primary antibody or through a labeled secondary antibody, that the problem arises.
The term immunofluorescence is then used to label this technique. However, this does not reveal the sample type, only the method of detection that was used.
Primary antibodies are often designated by biotech companies as being tested for use in immunofluorescence. This does not tell us very much. When choosing a product based on whether they have been validated for use in a particular application, the most important thing to consider is whether the product has been validated for the particular sample type in use (i.e. tissue or cells). The detection system (-chemistry or -fluorescence) comes secondary.
In this sense, immunofluorescence as an application term for primary antibodies is particularly useless. If it were possible to replace “-chemistry” with “-fluorescence” at the end of each term then the visualization method used would be much better indicated, whilst maintaining the sample type information.
Proposed Nomenclature for Immunostaining Techniques
Source: StressMarq Biosciences
| . |
Tissue (-histo) |
Cells (-cyto) |
| Chemical detection (-chemistry) |
Immunohistochemistry |
Immunocytochemistry |
| Fluorescent detection (-fluorescence) |
Immunohistofluorescence |
Immunocytofluorescence |
Key Definitions
Immunochemistry (IC) Definition
The use of an antibody and subsequent visualization of a chemical reaction that produces a color change in order to detect a target.
Immunohistochemistry (IHC) Definition
The use of an antibody and subsequent visualization of a chemical reaction that produces a color change in order to detect a target in tissue.
Immunocytochemistry (ICC) Definition
The use of an antibody and subsequent visualization of a chemical reaction that produces a color change in order to detect a target in cells.
Immunofluorescence (IF) Definition
The use of an antibody and subsequent visualization using a fluorophore in order to detect a target.
Immunohistofluorescence (IHF) Definition
The use of an antibody and subsequent visualization using a fluorophore in order to detect a target in tissue.
Immunocytofluorescence (ICF) Definition
The use of an antibody and subsequent visualization using a fluorophore in order to detect a target in cells.
It is apparent from Google searches that it will be a while before these terms are used as the norm.
The term immunofluorescence (IF) is still widely used and so, as such, it has been reluctantly grouped with the immunocytochemistry (ICC) application term here, as those two techniques are often combined.
The ICC/IF abbreviation is used throughout our website as a result. If an antibody is labeled as being tested for use in ICC/IF this means that it has been validated using cells with a secondary antibody conjugated to either a fluorescent or enzyme/protein tag.
The abbreviation IHC for immunohistochemistry means that the antibody may be used in tissue with a secondary antibody conjugated to any tag, as the binding of the primary antibody is not affected.
Acknowledgments
Produced from materials originally authored by Leslie Rietveld from StressMarq Biosciences Inc.
About StressMarq Biosciences
Established in 2007, StressMarq Biosciences Inc. is a supplier of life science products that operates out of Victoria, Canada with a small, but dedicated group of scientists. Headed by our CEO and President Dr. Ariel Louwrier, StressMarq provides the research community with high-quality reagents backed with rigorous quality control data, expert scientific support, and fast international delivery.
“Discovery through partnership, Excellence through quality”
With over 7,000 products, our growth can be attributed to the continual production of cutting edge research products. Our diverse portfolio of primary antibodies, antibody conjugates, proteins, immunoassay kits and small molecules bridges across the life sciences, including products for cancer research, cardiovascular disease, cell signaling and neuroscience. To aid research worldwide, StressMarq has an extensive network of international distributors that allow us to supply reagents to over 50 countries.
In the years to come, StressMarq will continue to aid life science research by providing “Discovery through partnership, and Excellence through quality”.
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