Macropinocytosis Assessment
Macropinocytosis Assessment - Related Videos
06:47
High-throughput Measurement of Dictyostelium discoideum Macropinocytosis by Flow Cytometry
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Cited by 8 •
2018
Macropinocytosis, large-scale non-specific fluid uptake, is important in many areas of clinical biology including immunology, infection, cancer, and neurodegenerative diseases. Here, existing techniques have been adapted to allow high-throughput, single-cell resolution measurement of macropinocytosis in the macropinocytosis model organism Dictyostelium discoideum using flow cytometry.
08:31
Fluorescence Microscopy for ATP Internalization Mediated by Macropinocytosis in Human Tumor Cells and Tumor-xenografted Mice
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Cited by 3 •
2021
We developed a reproducible method to visualize the internalization of nonhydrolyzable fluorescent adenosine triphosphate (ATP), an ATP surrogate, with high cellular resolution. We validated our method using independent in vitro and in vivo assays-human tumor cell lines and immunodeficient mice xenografted with human tumor tissue.
08:36
Neurotoxicity Assessment in Adult Danio rerio using a Battery of Behavioral Tests in a Single Tank
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Cited by 3 •
2023
Here, we present a comprehensive behavioral test battery, including the novel tank, Shoaling, and social preference tests, to effectively determine the potential neurotoxic effects of chemicals (e.g., methamphetamine and glyphosate) on adult zebrafish using a single tank. This method is relevant to neurotoxicity and environmental research.
06:50
Systematic Assessment of Well-Being in Mice for Procedures Using General Anesthesia
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Cited by 25 •
2018
We developed a protocol to assess well-being in mice during procedures using general anesthesia. A series of behavioral parameters indicating levels of well-being as well as glucocorticoid metabolites were analyzed. The protocol can serve as a general aid to estimate the degree of severity in a scientific, animal-centered manner.
07:08
Assessing and Flushing a Peripheral Intravenous Line
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2023
Source: Madeline Lassche, MSNEd, RN and Katie Baraki, MSN, RN, College of Nursing, University of Utah, UT After peripheral intravenous (IV) access is initiated, it is important to assess and maintain the IV catheter according to institutional policies and nursing standards of practice. The regular assessment of the insertion site and the surrounding areas for signs of complications is necessary to prevent IV catheter complications, including infiltration, phlebitis, infection, extravasation, or...
FAQs
Actin-driven membrane ruffles shape the earliest measurable stage of the process. When these folds move back onto the cell surface, they capture surrounding fluid and close into intracellular macropinosomes. An assessment therefore reflects both membrane remodeling and subsequent compartment formation, allowing changes in cytoskeletal activity or vesicle production to be examined through tracer uptake.
Fluorescent dextran serves as a tracer for extracellular fluid and dissolved material taken into the cell. After uptake, its fluorescence can be measured to compare macropinocytic activity between samples. Differences in tracer signal under controlled conditions can indicate altered internalization and help researchers examine how signaling pathways regulate this dynamic membrane-trafficking process.
Controlled conditions make tracer uptake comparisons interpretable. If samples differ in their experimental conditions, a change in fluorescence may not reflect regulation of macropinocytosis itself. Keeping relevant conditions consistent allows researchers to associate differences in dextran uptake with changes in cellular signaling or biological state, rather than with uncontrolled variation between measurements.
Microscopy and flow cytometry provide complementary ways to quantify fluorescent tracer uptake. Microscopy can measure fluorescence while retaining visual information about cells and intracellular compartments. Flow cytometry measures fluorescence across analyzed cells, supporting comparison of uptake within a sample population. Choosing between them depends on whether spatial observation or population-level measurement is more important.
A basic workflow exposes cells to a fluorescent tracer such as dextran, maintains comparable conditions across samples, and then measures intracellular fluorescence by microscopy or flow cytometry. The resulting uptake measurements are compared between conditions. This approach connects tracer accumulation with macropinocytic activity and can reveal how cellular signaling or state changes the process.
This assessment is useful when researchers need to characterize fluid and dissolved-material uptake in different biological contexts. It can support studies of nutrient acquisition, membrane trafficking, and immune-cell activity, while also revealing changes associated with cancer or infection. Comparing uptake patterns helps place these observations within broader studies of cellular regulation and disease-related biology.