What is the Difference Between Immunofluorescence and Immunohistochemistry?

Immunofluorescence (IF) and Immunohistochemistry (IHC) are molecular assays that involve the use of antibodies to detect specific proteins in tissue samples. Both techniques share similar principles, but they differ in the method of visualizing the proteins and the type of samples they are commonly used on.

Key differences between immunofluorescence and immunohistochemistry include:

Visualization method: IHC uses enzymatic labels, such as peroxidase and alkaline phosphatase, to visualize the antigens of interest, while IF uses fluorescent signals.
Microscope requirement: IHC is viewed with a brightfield microscope, whereas IF requires a fluorescence microscope.
Sample type: IF is commonly used to stain microbiological cells, while IHC is used to stain sections of biological tissue.
Staining permanence: IHC staining is permanent and does not fade over time, whereas the fluorescent signal in IF can fade over time, a phenomenon known as photobleaching.
Multiplexing: IHC can be easily multiplexed with chromogenic detection methods, allowing for the simultaneous detection of multiple proteins. In contrast, IF requires the use of multiple fluorochromes for multiplexing, which can be more challenging and lead to issues such as spectral overlap.

The choice between IF and IHC depends on the specific research question and the resources available. For example, if multiple proteins need to be detected simultaneously with higher spatial resolution, IF might be the more suitable choice. On the other hand, if the study requires higher throughput or more permanent staining, IHC could be more appropriate.

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What is the Difference Between Immunofluorescence and Immunohistochemistry?

Comparative Table: Immunofluorescence vs Immunohistochemistry

Here is a table comparing the differences between immunofluorescence and immunohistochemistry:

Technique	Immunofluorescence (IF)	Immunohistochemistry (IHC)
Definition	A technique that uses fluorescent dyes to visualize the presence of specific proteins or antigens in tissue samples.	A technique that uses chromogenic or fluorescent detection to visualize the presence of specific proteins or antigens in tissue samples.
Visualization	Fluorescent dyes are used to label the antibodies, which then bind to the target proteins or antigens, allowing them to be visualized under a fluorescence microscope.	Chromogenic detection with a colored or fluorescent dye is used to visualize the antibody-antigen interaction.
Sensitivity	Generally more sensitive than immunohistochemistry, as fluorescent dyes can produce brighter signals.	Generally less sensitive than immunofluorescence, as chromogenic detection may produce weaker signals.
Quantitation	Less quantitative than assays such as western blotting or ELISA, but provides valuable information about protein localization in the context of intact tissue.	Useful for studying protein expression patterns and pathological features in tissue samples.
Applications	Widely used in various research fields, such as cell biology, immunology, and neurobiology.	Commonly used for diagnostic and prognostic purposes in pathology.

Both immunofluorescence and immunohistochemistry are essential tools for studying protein expression and localization in tissue samples, but they differ in sensitivity, visualization methods, and applications.

Guilherme Mazui

Guilherme Mazui is graduated in journalism from the Federal University of Minas Gerais (UFMG) and a master's degree in Communication from the University of São Paulo (USP). In addition, he has experience in advertising writing and has worked as a content editor in several companies.