Optimum signal for brightness/sensitivity
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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 |  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 |
Suboptimal conditions may result in signal too weak. Additional optimization may be needed. |
|  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 |
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 |
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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.
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.
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.
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.
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.
Related products
Spatial biology
Find publications, protocols, and technical guides to start or evolve your multiplex imaging experiments and research.
Learn more at the Spatial Biology Resource Center
Spatial biology protocols:
- IHC FFPE Tissue Low pH Antigen Retrieval—Direct Method
- IHC FFPE Tissue High pH Antigen Retrieval—Direct Method
- Multiplexed SuperBoost Tissue Staining with Alexa Fluor Tyramides
Spatial biology poster:
Imaging protocols
Protocols that fit your needs in imaging ranging from sample and assay preparation to staining, labeling, and data analysis strategies.
Protocols include:
- Cell proliferation, viability, structure
- Live cell assays
- Immunolabeling
- Cell painting
- Spatial biology and multiplex IHC
Additionally, search through our library of online microscopy protocols
Imaging tools
Find helpful tools below to help you plan your imaging experiments.
Stain-iT Cell Staining Simulator Visualize staining your cell without wasting your reagents, antibodies, or time.
Resources
For Research Use Only. Not for use in diagnostic procedures.
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