Nanoscale effects of ethanol and naltrexone on protein organization in the plasma membrane studied by photoactivated localization microscopy (PALM) (CAT#: STEM-MIT-0398-LJX)

Introduction

Ethanol affects the signaling of several important neurotransmitter and neuromodulator systems in the CNS. It has been recently proposed that ethanol alters the dynamic lateral organization of proteins and lipids in the plasma membrane, thereby affecting surface receptor-mediated cellular signaling. Our aims are to establish whether pharmacologically relevant levels of ethanol can affect the lateral organization of plasma membrane and cytoskeletal proteins at the nanoscopic level, and investigate the relevance of such perturbations for mu-opioid receptor (MOP) function.
The service uses Photoactivated Localization Microscopy with pair-correlation analysis (pcPALM), a quantitative fluorescence imaging technique with high spatial resolution (15-25 nm) and single-molecule sensitivity, to study ethanol effects on protein organization in the plasma membrane.

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Principle Applications Procedure Materials Notes Related Products

Principle

Principles of photoactivated localization microscopy: By fitting the two-dimensional Gaussian function to determine the centroid of microscope-formed light spots, a single fluorescent source (such as a fluorescent group) can be located with high precision. The accuracy of the calculation to determine the centroid depends only on the number of photons collected, and the resolution scale can be tens of nanometers or smaller. To achieve this accuracy, the density of the fluorescent molecules being tested is required to be low enough that the spots of the two fluorescent groups are unlikely to overlap.

Applications

Applied in many areas of the life sciences,

Procedure

1. Sampling
2. Preparation of slices
3. Staining (Select according to the specific experimental situation)
4. Observation

Materials

• Sample Type:
Protein tissue in the plasma membrane

Notes

Operate in strict accordance with the operating procedures, and shall not arbitrarily change the operating procedures

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