Intracellular Marker Labeling
Intracellular Marker Labeling - Related Videos
08:34
Identification of Virulence Markers of Mycobacterium abscessus for Intracellular Replication in Phagocytes
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Cited by 5 •
2018
Here, we present two protocols to study Phagocyte-Mycobacterium abscessus interactions: the screening of a transposon mutant library for bacterial intracellular deficiency and the determination of bacterial intracellular transcriptome from RNA sequencing. Both approaches provide insight into the genomic advantages and transcriptomic adaptations enhancing intracellular bacteria fitness.
08:14
Detection of Intracellular Gene Expression in Live Cells of Murine, Human and Porcine Origin Using Fluorescence-labeled Nanoparticles
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Cited by 1 •
2015
This manuscript describes a novel technique which allows the detection of intracellular gene expression after endocytosis of fluorescence-labeled nanoparticles directly in live cells. The method does not require manipulation of the cells and is not restricted with respect to the target gene or species.
03:20
Labeling Neural Cell Surface Markers with Azidosugar while Co-culturing with Endothelial Cells
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2025
This video demonstrates a protocol for co-culturing mouse brain endothelial cells and primary cortical stem cells and labeling the cell surface glycoproteins using Ac4ManNAz (azidosugar per-O-acetylated N-azidoacetylmannosamine). This method allows for selective enrichment and identification of cell surface proteins in primary cells.
10:30
The C. elegans Excretory Canal as a Model for Intracellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis in a Single Cell: labeling by GFP-fusions, RNAi Interaction Screen and Imaging
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Cited by 6 •
2017
The C. elegans excretory canal is a unique single-cell model for the visual in vivo analysis of de novo polarized membrane biogenesis. This protocol describes a combination of standard genetic/RNAi and imaging approaches, adaptable for the identification and characterization of molecules directing unicellular tubulogenesis, and apical membrane and lumen biogenesis.
07:18
Intracellular Refolding Assay
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Cited by 9 •
2012
In this protocol a method to measure intracellular protein refolding after heat shock is described. This method can be used to study foldases like molecular chaperones and their co-factors or compounds able to influence their activity. Firefly luciferase activity is used as reporter to measure chaperone refolding activity.
FAQs
Fixation stabilizes cellular contents before permeabilization, creating a preserved sample in which intracellular targets can be labeled. Permeabilization then makes the plasma membrane sufficiently accessible to antibodies or fluorescent probes. The sequence matters because labeling requires access to internal molecules while retaining the cellular information needed for later flow-cytometric or microscopic analysis.
Surface markers describe molecules exposed at cell boundaries, but important functional or regulatory information may be inside the cell. Measuring intracellular cytokines, transcription factors, or pathogen antigens adds evidence about what a cell is producing, regulating, or carrying. Combining these measurements with cellular identification can therefore distinguish cells with different response states within the same population.
Individual-cell measurements show whether a marker is distributed across many cells or concentrated in a subset. This distinction is important in complex immune populations, where an average signal could conceal functionally different groups. In infection studies, examining intracellular pathogen antigens or response-associated proteins at the cell level can connect infection-related changes with specific cellular states rather than only a bulk population result.
A typical workflow begins by preserving cells through fixation, followed by permeabilization and exposure to antibodies or fluorescent probes directed toward selected intracellular targets. The labeled cells can then be examined by flow cytometry for measurements across individual cells or by microscopy. The chosen targets should match the biological question, such as immune function, transcriptional regulation, or infection-associated antigen detection.
Researchers use intracellular marker labeling when the question concerns immune-cell function or infection-related changes that cannot be addressed with surface information alone. It supports comparisons among different treatment conditions when the goal is to determine how cellular responses differ. Because measurements can be made in individual cells, the method is also useful for identifying heterogeneous response patterns in complex populations.
Flow cytometry and microscopy can turn intracellular labels into evidence about cellular state and response. Depending on the selected marker, results may indicate cytokine-associated activity, transcriptional regulation, pathogen antigen carriage, or other response-associated changes. Comparing these signals across immune-cell groups or experimental treatments helps researchers trace infection-related effects and determine whether responses are shared broadly or restricted to subsets.