Immunofluorescence Imaging

What is immunofluorescence imaging?

Immunofluorescence imaging is a powerful technique used in biological research to visualize the distribution, localization, and interaction of specific proteins within cells and tissues. This technique uses fluorescently labeled antibodies to target proteins with high specificity, enabling researchers to visualize the spatial organization of proteins and better understand their role in complex cellular pathways. By combining the specificity of antibodies with the detection capabilities of fluorescence microscopy, researchers can precisely localize and quantify proteins within individual structures. Immunofluorescence imaging not only provides valuable insights into subcellular protein localization but also the levels of target abundance. Additionally, this imaging technique contributes to our understanding of cellular processes, such as protein trafficking and localization changes in response to various stimuli or cellular conditions. Immunofluorescence imaging has thus become an indispensable tool for researchers seeking to unravel the complexities of cellular biology and disease mechanisms.

Thermo Fisher Scientific offers a variety of immunofluorescence imaging tools for any level of target abundance (high, medium, low). Below we summarize the optimal tools (primary, secondary, and streptavidin conjugation, along with tyramide signal amplification and Aluora signal amplification) available for each level of abundance:

	Conjugated primary antibodies	Secondary antibody detection	Streptavidin-based amplification	Tyramide signal amplification	Aluora spatial amplification
High-abundance target (e.g., tubulin)	Optimum signal for brightness/sensitivity	Optimum signal for brightness/sensitivity	Suboptimal conditions may result in signal too strong. Additional optimization may be needed.	Suboptimal conditions may result in signal too strong. Additional optimization may be needed.	Suboptimal conditions may result in signal too strong. Additional optimization may be needed.
Medium-abundance target (e.g., Golgi or mitochondria)	Suboptimal conditions may result in signal too weak. Additional optimization may be needed.	Optimum signal for brightness/sensitivity	Optimum signal for brightness/sensitivity	Suboptimal conditions may result in signal too strong. Additional optimization may be needed	Suboptimal conditions may result in signal too strong. Additional optimization may be needed
Low-abundance target (e.g., receptors or cell junction)	Suboptimal conditions may result in signal too weak. Additional optimization may be needed.	Suboptimal conditions may result in signal too weak. Additional optimization may be needed.	Optimum signal for brightness/sensitivity	Optimum signal for brightness/sensitivity	Optimum signal for brightness/sensitivity in high-plex tissue imaging

Conjugated primary antibodies

Secondary antibody detection

Streptavidin-based amplification

Tyramide signal amplification

Aluora spatial amplification

High-abundance target
(e.g., tubulin)

Optimum signal for brightness/sensitivity

Suboptimal conditions may result in signal too strong. Additional optimization may be needed.

Medium-abundance target
(e.g., Golgi or mitochondria)

Suboptimal conditions may result in signal too weak. Additional optimization may be needed.

decorative image
Optimum signal for brightness/sensitivity

Optimum signal for brightness/sensitivity

Suboptimal conditions may result in signal too strong. Additional optimization may be needed

Low-abundance target
(e.g., receptors or cell junction)

Suboptimal conditions may result in signal too weak. Additional optimization may be needed.

Optimum signal for brightness/sensitivity

decorative image Optimum signal for brightness/sensitivity

decorative image
Optimum signal for brightness/sensitivity in high-plex tissue imaging

multiple cell images with different fluorescent primary antibodies

Conjugated Primary Antibodies

Direct detection of your target with ease

Primary antibodies selectively bind to target antigens, offering high specificity and sensitivity. We offer a wide variety of primary antibodies against key signaling proteins, cellular markers, and organelles, as well as cell junctions and CD markers.

Learn more about conjugated primary antibodies

Secondary Antibodies

Provide that extra signal and assay flexibility

Secondary antibodies amplify the signal and allow for more sensitive detection of the target antigen. They are essential in various immunological techniques, such as Western blotting, immunofluorescence, and ELISA, to improve the specificity and sensitivity of the assay.

Learn more about secondary antibodies

fluorescent cell with green streptavidin Golgi stain

Streptavidin Signal Amplification

Boost the signal of medium- and low-abundance targets

For improved detection sensitivity, streptavidin-based amplification techniques are widely used in fluorescence imaging to detect primary and secondary antibodies, ligands, and toxins, or DNA probes for in situ hybridization.

Learn more about streptavidin conjugates

Tyramide Signal Amplification

Simple amplification for low-abundance targets

With SuperBoost tyramide signal amplification technology, you can see and measure low-abundance targets that are not detectable by conventional means. Tyramide-based signal amplification provides excellent sensitivity without compromising resolution.

Learn more about TSA signal amplification

Aluora Spatial Amplification

Spatial amplification method for low-abundance multiplex detection in IHC

Aluora spatial amplification allows for the detection of spatial relationships and cellular interactions in tissue samples. Aluora reagents offer enhanced brightness and improved sensitivity for the detection of multiple targets simultaneously in one tissue sample and help overcome autofluorescence when labeling rare targets.

Learn more about Aluora spatial amplification