Immunocytochemical Analysis
Immunocytochemical Analysis - Related Videos
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.
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.
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.
15:04
A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity
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Cited by 46 •
2009
This article describes a novel protocol and reagent set designed for sensitive measurement of neurotoxic effects of compounds and treatments on co-cultures of neurons and astrocytes using high content analysis. Results demonstrate that high content analysis represents an exciting novel technology for neurotoxicity assessment.
FAQs
The primary antibody provides the target-recognition step: it binds a selected protein or other antigen with high specificity. A labeled secondary antibody then makes that bound primary antibody visible, whereas a directly tagged probe places the label on the detecting reagent itself. This choice determines how the target signal is generated for microscopy.
Direct detection uses a tagged probe to identify the target, while secondary-antibody detection uses a primary antibody followed by a labeled secondary antibody. The two formats differ in where the visual label is attached, so researchers can select the arrangement that fits the target-detection design and the intended microscopy readout.
The observed pattern provides spatial information within preserved cellular structures. In neural samples, researchers can examine whether neuronal markers, neurotransmitter-related proteins, receptors, or glial components occur in particular cells or regions. Comparing these patterns helps characterize cell identity, neural development, connectivity, disease-related changes, or responses to experimental treatments.
Preserved cellular structures maintain the spatial setting in which the signal is observed. That context allows microscopy to associate a target with individual cells and their surrounding neural organization rather than treating the sample as an undifferentiated mixture. This is particularly relevant when examining neuronal markers, receptors, neurotransmitter-related proteins, or glial components.
A typical workflow begins with preserved cultured cells or tissue sections, followed by exposure to a primary antibody. Detection then uses either a labeled secondary antibody or a directly tagged probe, and microscopy is used to examine the resulting signal. The workflow connects molecular recognition with cellular and tissue-level visualization.
The method can be applied to cultured cells and tissue sections, allowing questions at different organizational scales. Cultured cells support examination of cellular markers in prepared cell systems, whereas sections retain tissue context for evaluating distributions across neural structures. In both settings, targets may include neuronal, neurotransmitter-related, receptor, or glial components.
It is useful when researchers need to relate molecular markers to neural organization or experimental change. Applications include identifying cell types, examining neural development and connectivity, characterizing disease-related alterations, and assessing responses to treatments. Mapping glial components and neurotransmitter-related proteins can add cellular context to these investigations.
Differences in signal location, abundance, or distribution can be compared between neural samples when studying disease-related changes or treatment responses. Such comparisons help determine whether a target-associated pattern differs across conditions, while the cellular context indicates which neuronal or glial components are involved. The method therefore links molecular observations with broader neuroscience questions.