Extracellular Staining
Extracellular Staining - Related Videos
04:33
Extracellular DNA Staining: A Method to Identify Extracellular DNA in FFPE Tissue Sections
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2023
This video describes the technique of staining extracellular DNA in FFPE tissue sections. The stained extracellular DNA can be quantitated to determine the efficacy of any therapeutic treatment that leads to the death of cancer cells.
04:29
Immunofluorescent Staining of Spheroids: A Technique to Analyze Spheroids for Expression of Intra- and Extracellular Antigen Expression
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2023
This video describes the technique of immunofluorescent staining of spheroids to understand the differentiation of the fibroblasts or CAFs. This protocol helps detect expression of intra- and extracellular antigens, including deposition of ECM proteins within the spheroids.
11:33
Microscopy and Staining: Gram, Capsule, and Endospore Staining
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2023
Source: Rhiannon M. LeVeque1, Natalia Martin1, Andrew J. Van Alst1, and Victor J. DiRita1 1 Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, United States of America Bacteria are diverse microorganisms found nearly everywhere on Earth. Many properties help distinguish them from each other, including but not limited to Gram-staining type, shape and arrangement, production of capsule, and formation of spores. To observe these properties, one...
09:39
Techniques for the Analysis of Extracellular Vesicles Using Flow Cytometry
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Cited by 31 •
2015
Many different methods exist for the measurement of extracellular vesicles (EVs) using flow cytometry (FCM). Several aspects should be considered when determining the most appropriate method to use. Two protocols for measuring EVs are presented, using either individual detection or a bead-based approach.
14:17
Neutrophil Extracellular Traps: How to Generate and Visualize Them
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Cited by 296 •
2010
Neutrophil Extracellular Traps (NETs) are an important innate immune mechanism to fight pathogenic bacteria, fungi and parasites. Here we describe methods to isolate neutrophil granulocytes from human blood and to activate them to form NETs. We present preparation techniques to visualize NETs in light and electron microscopy.
FAQs
Membrane preservation is essential because the targets being measured remain accessible only when their extracellular location and the plasma membrane are maintained. Antibodies conjugated to fluorescent labels can then bind those exposed antigens without relying on signals from inside the cell. This supports more reliable identification of surface-defined populations in immune samples.
The choice of fluorescently conjugated antibodies determines which surface antigens can be measured and how many markers can be examined together. In multiparameter extracellular staining, each antibody contributes a separate fluorescence signal, allowing combinations of markers to distinguish immune-cell populations or assess activation and differentiation states. The resulting pattern is more informative than measuring a single antigen alone.
Viability dyes and sample-processing steps address two different sources of uncertainty. A viability dye helps distinguish cells according to viability, while processing intended to reduce nonspecific binding helps limit fluorescence that does not arise from the selected antibody-antigen interaction. Including both considerations makes population measurements easier to interpret, particularly in complex immunology and infection samples.
A basic workflow starts with sample processing, applies fluorescently conjugated antibodies under conditions that preserve the plasma membrane, and incorporates measures to reduce nonspecific binding. Viability dyes may be included before the labeled sample is analyzed by flow cytometry or fluorescence microscopy. Keeping these stages distinct helps connect sample preparation with the quality of the final signal.
Extracellular staining can profile immune-cell populations, measure activation or differentiation markers, and examine receptors relevant to host-pathogen interactions. In infection studies, repeating measurements across experimental conditions can reveal shifts in cellular responses. The same strategy supports comparisons during vaccination or treatment, where marker patterns help track biological changes over time or between groups.
It enables researchers to examine receptors exposed on cells that participate in host-pathogen interactions while placing those receptors in a broader cellular profile. Combining receptor measurements with activation, differentiation, or population markers can show which cell groups change during infection. This links surface phenotype to immune responses without requiring the analysis to focus on a single marker.