Macropinocytosis Quantification
Macropinocytosis Quantification - 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.
06:58
Quantification of Membrane Ruffle Formation Using Scanning Electron Microscopy
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2025
This video demonstrates sample preparation, fixation, and imaging of a macrophage cell that displays membrane ruffles using scanning electron microscopy, SEM. Detailed information about membrane morphology and organization is crucial for understanding the physiological condition of the cells.
08:44
Invertebrate Lifespan Quantification
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2023
Many animals naturally stop growing upon reaching adulthood, after which they undergo aging or "senescence" until dying. The amount of time between an organism\'s birth and death is called its lifespan, which can be influenced by various biological and environmental factors. By exposing organisms to different growth conditions, scientists can better understand the factors affecting lifespan. Flies and worms are ideal organisms to perform such experiments, given their short generation time and...
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.
08:38
Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS
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Cited by 28 •
2015
Here we describe a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay to quantify the immunosuppressant tacrolimus in dried blood spots using a simple manual protein precipitation step and online column extraction.
FAQs
Actin-driven membrane ruffling creates folds that move back onto the cell surface and enclose extracellular fluid. These events generate macropinosomes, so changes in ruffle formation can alter the amount of tracer entering cells. Measuring uptake therefore provides information about cytoskeletal membrane activity as well as the resulting vesicle population, rather than representing a simple measurement of extracellular fluid exposure.
A fluorescence-based uptake measurement can be interpreted alongside the stages that produce the signal. Differences may arise from altered macropinosome formation, changes in the amount of dissolved tracer enclosed, or subsequent intracellular trafficking. This distinction matters because similar overall uptake values can conceal different cellular behaviors, while comparative measurements can help identify which part of the process changes.
Fluorescent dextran and related fluid-phase tracers provide a detectable signal associated with extracellular fluid enclosed during uptake. Their fluorescence allows researchers to estimate how much fluid-phase material cells internalize without relying only on visualizing membrane shape. The resulting measurement is especially useful for comparing uptake between cell types, treatments, or genetic conditions.
A typical measurement uses a fluorescent dextran or another fluid-phase tracer, followed by detection of cellular fluorescence. Microscopy records tracer-associated signals in individual cells and can relate them to visible cellular structures, whereas flow cytometry measures fluorescence across many cells. Selecting either approach depends on whether the comparison emphasizes cellular imaging or population-level uptake.
Comparative measurements are useful when a study asks whether cellular context changes bulk fluid uptake. Researchers can examine differences among cell types, assess the effect of treatments, or evaluate genetic conditions that may modify the process. Because the same quantification framework supports these comparisons, it can reveal relative changes in uptake efficiency under defined biological conditions.
Macropinocytosis quantification supports investigations of nutrient acquisition, immune-cell function, pathogen entry, and cancer-cell metabolism. In each setting, the measurement links fluid uptake with a broader biological question, such as how cells obtain dissolved materials or respond to their environment. It also provides a way to compare uptake behavior across experimental groups rather than describing membrane activity qualitatively.