Visualizing Membrane Ruffle Formation using Scanning Electron Microscopy

作者： WonMo Ahn1, Bhupesh Singla1, Brendan Marshall2, Gábor Csányi1,3 1Vascular Biology Center,Medical College of Georgia at Augusta University, 2Department of Cellular Biology and Anatomy,Medical College of Georgia at Augusta University, 3Department of Pharmacology and Toxicology,Medical College of Georgia at Augusta University

简介：

Overview

This protocol outlines a method to visualize and quantify membrane ruffle formation and macropinocytic activity using scanning electron microscopy (SEM). It highlights the importance of macropinocytosis in various pathological conditions.

Key Study Components

Area of Science

• Cell Biology
• Neuroscience
• Microscopy Techniques

Background

• Macropinocytosis is a conserved endocytic process.
• It involves the formation of membrane ruffles driven by F-actin.
• Increased macropinocytosis is linked to various diseases.
• SEM provides high-resolution imaging of cellular structures.

Purpose of Study

• To visualize membrane ruffles and macropinocytic cups.
• To quantify membrane ruffle formation in vitro.
• To identify signaling pathways regulating macropinocytosis.

Methods Used

• Preparation of sterile glass cover slips in a 24-well plate.
• Seeding of RAW 264.7 macrophages at a specified density.
• Incubation of cells in a humidified environment.
• Use of SEM for imaging membrane activities.

Main Results

• High-resolution images of membrane ruffles were obtained.
• Quantification of macropinocytic activity was achieved.
• Potential signaling pathways were identified.
• Effects of inhibitors on macropinocytosis were assessed.

Conclusions

• SEM is an effective tool for studying macropinocytosis.
• The protocol can help discover new regulators of this process.
• Understanding macropinocytosis may lead to insights into disease mechanisms.

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