Macrophage Isolation
Macrophage Isolation - Related Videos
07:46
Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
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Cited by 15 •
2017
This protocol allows a researcher to isolate and characterize tissue-resident macrophages in various hallmark inflamed tissues extracted from diet-induced models of metabolic disorders.
09:25
Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
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Cited by 68 •
2012
Here, we detail a methodology for the rapid isolation of mouse intestinal dendritic cells (DCs) and macrophages. Phenotypic characterization of intestinal DCs and macrophages is performed using multi-color flow cytometric analysis while magnetic bead enrichment followed by cell sorting is used to yield highly pure populations for functional studies.
03:37
Isolation of Phagosomes Containing Magnetic Bead-Tagged Bacteria from Infected Macrophages
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2026
Source: Gutiérrez, S., et al., Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages. J. Vis. Exp. (2017)This video demonstrates the magnetic isolation of phagosomes containing biotin–streptavidin bead–coated bacteria from infected macrophages. The protocol includes buffer treatments, cell disruption, magnetic separation, and washing to yield purified phagosomes for analysis.
06:56
Isolation of Macrophages from Mouse Dorsal Root Ganglion Using Mechanical Dissociation
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2025
This video demonstrates an enzyme-free protocol to isolate macrophages from mouse dorsal root ganglion (DRG) using a mechanical dissociation technique. Upon homogenization of the freshly collected DRG tissue, antibodies against the cell-surface receptors on the macrophages are used to label the cells fluorescently, followed by performing flow cytometry to confirm the presence of macrophages in the sample.
09:04
Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy
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Cited by 3 •
2014
A method to evaluate the ability of isolated mouse alveolar macrophages to control the growth of phagocytosed Aspergillus spores by confocal microscopy.
FAQs
Each enrichment strategy selects cells according to a different property. Density-gradient centrifugation separates cells by density, adherence-based separation uses attachment behavior, magnetic-activated cell sorting relies on labeled surface markers, and flow cytometry identifies or sorts marker-defined populations. The choice determines whether the experiment prioritizes broader macrophage recovery or a more specifically characterized subset.
Mechanical or enzymatic dissociation releases macrophages from the surrounding tumor tissue so they can undergo enrichment. This initial step connects the physical structure of a tumor sample with downstream separation methods, including density gradients, magnetic sorting, or flow cytometry. Its role is especially important when investigators need to examine macrophages that occur within, rather than outside, the tumor environment.
Isolated cells provide a preparation for examining macrophage features linked to tumor biology, including cytokine production, immune suppression, phagocytosis, and interactions with cancer cells. Investigators can use these functional readouts to compare macrophage phenotypes or polarization states. This makes isolation valuable for connecting cellular characteristics with the behavior of macrophages in the tumor microenvironment.
A typical workflow begins with collecting macrophages from blood, tissue, a tumor, or a culture, followed by mechanical or enzymatic dissociation when necessary. The resulting cell preparation then undergoes selective enrichment using density-gradient centrifugation, adherence, magnetic-activated cell sorting, or flow cytometry based on the experimental goal. The isolated population is subsequently used for biological or functional analysis.
Researchers use isolated macrophages when they need to study immune activity within the tumor microenvironment or test how macrophages interact with cancer cells. The approach supports investigations of tumor immunity, macrophage polarization, cytokine production, immune suppression, and phagocytosis. It also creates a defined cellular material for comparing macrophage states across experiments or evaluating interventions that alter tumor-associated immune behavior.
Macrophage preparations can support biomarker discovery and the development or testing of therapies designed to alter the tumor microenvironment. Functional measurements, such as cytokine production, phagocytosis, immune suppression, or interactions with cancer cells, provide information about macrophage behavior. These outcomes help relate cellular phenotypes to tumor immunity and assess whether an intervention changes macrophage-associated responses.