Immunofluorescence Profiling
Immunofluorescence Profiling - Related Videos
09:56
Immunofluorescence Microscopy: Immunofluorescence Staining of Paraffin-Embedded Tissue Sections
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2023
Source: Thomas Chaffee1, Thomas S. Griffith2,3,4, and Kathryn L. Schwertfeger1,3,4 1 Department of Lab Medicine and Pathology, University of Minnesota, Minneapolis, MN 55455 2 Department of Urology, University of Minnesota, Minneapolis, MN 55455 3 Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455 4 Center for Immunology, University of Minnesota, Minneapolis, MN 55455 Pathologic analyses of tissue sections can be used to obtain a better understanding of normal tissue...
09:12
High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research
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2025
This report describes an automated protocol for multiplex immunofluorescence (mIF) assays on an automated slide stainer. It demonstrates the high reproducibility of spatial proteomics on standard high-throughput tissue autostainers and whole-slide fluorescence imagers, which is ideal for custom mIF assays across a large number of tissue slides in translational research.
08:29
Expression Profiling with Microarrays
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2023
Microarrays are important tools for profiling gene expression, and are based on complementary binding between probes that are attached to glass chips and nucleic acids derived from samples. Using these arrays, scientists can simultaneously evaluate the expression of thousands of genes. In addition, the expression profiles of different cells or tissue types can be compared, allowing researchers to deduce how the expression of different genes change during biological processes, and thus gain...
07:13
Using Topographic Maps to Generate Topographic Profiles
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2023
Source: Laboratory of Alan Lester - University of Colorado Boulder Topographic maps are "plan-view" representations of Earth's three-dimensional surface. They are a standard type of map-view that provides an overhead, or aerial, perspective. Among the defining features of a topographic map are the contour lines that indicate locations of constant elevation. The elevation interval between the contour lines is dependent on the level of detail provided by the map and the kind of topography present.
01:12
Immunofluorescence Microscopy
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2023
A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange. The...
FAQs
Signal specificity comes from the primary antibody binding its target antigen. A fluorescent secondary antibody then binds the primary antibody and produces a visible signal, while directly conjugated labels attach the fluorescent marker to the antibody recognizing the target. Microscopy reveals where the signal occurs, allowing researchers to relate protein presence to cellular or tissue patterns.
Multiplexed staining allows several markers to be examined in the same biological sample. Comparing their fluorescence patterns can distinguish cell identities, assess multiple proteins, and reveal relationships among molecular signals and tissue organization. This broader profile is useful when a single marker cannot capture the cellular or disease-associated phenotype being investigated.
Localization shows where a protein is positioned within cells or tissues, not only how much is present. In genetics, comparing localization patterns can connect mutations or gene activity with altered cellular organization and protein distribution. These spatial changes may provide a more informative phenotype than abundance measurements alone when evaluating gene function.
The workflow centers on binding antibodies to target antigens, generating fluorescent signals through secondary antibodies or directly conjugated labels, and examining the sample with fluorescence microscopy. Researchers then compare signal location, pattern, and relative quantity across samples or conditions. This approach supports analysis of changes in proteins, cells, and molecular organization.
This technique can help investigate gene function, mutation-associated changes, disease-related phenotypes, and responses to experimental conditions. By examining protein abundance, localization, cell identity, and tissue organization, researchers can connect genetic differences or altered gene activity with observable molecular and cellular outcomes. Multiplexed measurements extend this analysis across several markers simultaneously.
Fluorescence microscopy provides spatially resolved patterns that can be localized and quantified. Depending on the markers selected, analysis may compare protein abundance, identify cells, assess molecular distributions, or evaluate tissue organization. These outcomes help determine whether an experimental condition or genetic change is associated with a measurable shift in cellular or molecular phenotype.