Sinusoid Leukocyte Isolation
Sinusoid Leukocyte Isolation - Related Videos
03:42
Isolation of Marginating Hepatic Leukocytes from a Mouse Liver
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2025
In this video, we demonstrate the isolation of marginating hepatic leukocytes from mouse liver by forced perfusion.
06:44
Isolation of Brain-infiltrating Leukocytes
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Cited by 33 •
2011
A rapid method to obtain infiltrating leukocytes from the murine brain is described. This method utilizes a continuous Percoll gradient and discontinuous Ficoll gradient to select and purify the leukocyte-enriched layer. Isolated leukocytes may then be characterized by flow cytometric measurements.
08:19
Isolation of Leukocytes from the Human Maternal-fetal Interface
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Cited by 57 •
2015
Described herein is a protocol to isolate and further study the infiltrating leukocytes of the decidua basalis and decidua parietalis - the human maternal-fetal interface. This protocol maintains the integrity of cell surface markers and yields enough viable cells for downstream applications as proven by flow cytometry analysis.
07:51
Isolation of Leukocytes from the Murine Tissues at the Maternal-Fetal Interface
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Cited by 44 •
2015
Described herein is a protocol to isolate and analyze the infiltrating leukocytes of tissues at the maternal-fetal interface (uterus, decidua, and placenta) of mice. This protocol maintains the integrity of most cell surface markers and yields enough viable cells for downstream applications including flow cytometry analysis.
06:53
Selective Harvesting of Marginating-hepatic Leukocytes
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Cited by 1 •
2016
Marginating-hepatic leukocytes exhibit unique characteristics and distinct immunological functions compared to other leukocyte populations. Here we describe a method for selective harvesting of this specific hepatic cell population, through forced perfusion of the liver of rats or mice. Marginating-hepatic leukocytes seem critical in determining susceptibility to hepatic-related diseases and metastases.
FAQs
Tissue perfusion helps recover immune cells located within or passing through tissue sinusoids before the tissue is processed further. In liver-based preparations, this step supports access to cells involved in local immune surveillance and infection control. Its role is therefore not merely preparative: it helps make the recovered leukocyte population more representative of the sinusoidal immune compartment.
The two treatments address different physical barriers within tissue. Enzymatic dissociation helps break down tissue structure, while mechanical disruption assists in releasing cells from the resulting material. Their combined use supports recovery of leukocytes from sinusoidal tissue while the procedure still aims to preserve cell viability, which is essential for downstream flow cytometry, microscopy, and functional assays.
Density-based centrifugation separates cellular material according to differences in density after tissue dissociation. This helps distinguish leukocytes from hepatocytes, debris, and other tissue components, producing a preparation more suitable for immune-cell analysis. The resulting fraction can contain macrophages, lymphocytes, and other myeloid cells, allowing researchers to examine several immune populations rather than a single cell type.
A useful preparation requires both effective tissue breakdown and preservation of leukocyte viability. Insufficient disruption may limit cell recovery, whereas processing that compromises viability can reduce the value of later analyses. The separation stage also needs to remove hepatocytes and debris sufficiently for interpretation. These linked considerations determine whether the recovered cells remain suitable for phenotypic or functional investigation.
A typical workflow begins with tissue perfusion, followed by enzymatic dissociation and mechanical disruption. The resulting suspension then undergoes density-based centrifugation to separate leukocytes from hepatocytes, debris, and other components. Researchers subsequently analyze the recovered cells using flow cytometry, microscopy, or functional assays. Together, these stages move from tissue access and release to enrichment and characterization.
Flow cytometry can be used to examine the recovered immune-cell populations, while microscopy provides cellular visualization. Functional assays add information about how the cells behave or respond under experimental conditions. Using these complementary approaches, researchers can study macrophages, lymphocytes, and other myeloid cells in relation to tissue-specific immunity, inflammation, infection control, or responses to therapeutic interventions.