Immunocytochemical Detection
Immunocytochemical Detection - Related Videos
09:56
Small-scale Propagation of Human iPSCs in Serum-free Conditions for Routine Immunocytochemical Characterization
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2017
Regular characterization of induced pluripotent stem cells (iPSCs), to ascertain maintenance of their pluripotent state, is an important step before these cells are used for other applications. Here we describe a method for the small-scale propagation of human iPSCs specifically designed to enable their easy and routine characterization via immunocytochemistry.
09:16
Immunocytochemical Analysis of Human Pluripotent Stem Cells using a Self-Made Cytospin Apparatus
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Cited by 1 •
2010
Suspension immunocytochemical staining of human pluripotent stem cells (hPSCs) for cell-surface markers (SSEA-3/SSEA-4) was achieved based on use of a self-made cytospin apparatus to create a monolayer of cells for observation and quantification.
10:27
Zika Virus Infection of Cultured Human Fetal Brain Neural Stem Cells for Immunocytochemical Analysis
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Cited by 2 •
2018
This article details the methods that are used to expand human fetal brain neural stem cells in culture, as well as how to differentiate them into various neuronal subtypes and astrocytes, with an emphasis on the use of neural stem cells to study Zika virus infection.
11:22
Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle
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Cited by 39 •
2015
The main adherent cell types derived from human muscle are myogenic cells and fibroblasts. Here, cell populations are enriched using magnetic-activated cell sorting based on the CD56 antigen. Subsequent immunolabelling with specific antibodies and use of image analysis techniques allows quantification of cytoplasmic and nuclear characteristics in individual cells.
09:10
Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce
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Cited by 17 •
2010
This protocol describes a simple adhesive-tape-based approach for sampling of tomato and other fresh produce surfaces, followed by rapid whole cell detection of Salmonella using fluorescence in situ hybridization (FISH).
FAQs
The primary antibody recognizes and binds the target antigen within the cell. A secondary antibody carrying either a fluorescent or enzymatic label then binds to the primary antibody, creating a signal associated with the target’s cellular location. This arrangement allows researchers to visualize where specific proteins occur while retaining information about their distribution among individual cells.
Fixation and permeabilization prepare cells for antibody-based analysis before the labeling steps begin. Together, they support access to intracellular target molecules so primary antibodies can interact with proteins inside the cell. In neural samples, this preparation is essential when the targets include cytoskeletal proteins, signaling molecules, neurotransmitter receptors, or markers located within the cellular interior.
Both labeling strategies make antibody-antigen binding detectable, but they produce signals through different readouts. Fluorescent labels support visualization through emitted fluorescence, whereas enzymatic labels generate a detectable reaction associated with the bound antibody. The choice therefore determines how the protein distribution is visualized and documented in the cells being examined.
A typical workflow begins by fixing and permeabilizing the cells, followed by exposure to a primary antibody directed against the protein of interest. A labeled secondary antibody is then applied to bind the primary antibody, and the resulting fluorescent or enzymatic signal is detected. The final observation reveals the target’s distribution within individual cells.
In neuroscience, the technique can be directed toward neurotransmitter receptors, neuronal markers, glial markers, cytoskeletal proteins, and signaling molecules. Examining these targets provides different types of cellular information, including features related to cell identity, structural organization, and signaling. The selected protein determines which aspect of neural cell biology the analysis emphasizes.
Researchers can compare the cellular distribution of selected proteins across developmental conditions or disease-related states. Changes in neuronal or glial markers may help characterize cell identity, while altered receptor, cytoskeletal, or signaling-protein patterns can reveal differences in cellular structure and function. The method therefore supports spatial comparisons of neural changes across experimental conditions.
Spatial measurements show where a target protein is located within individual cells and how its distribution changes after an experimental treatment. By examining receptors, markers, cytoskeletal proteins, or signaling molecules, investigators can assess treatment-associated differences in cellular structure or function. These observations provide cellular-level evidence that complements broader assessments of neural responses.