Quantitative Analysis of PcG-Associated Condensates by Stochastic Optical Reconstruction Microscopy (STORM) (CAT#: STEM-MIT-0384-LJX)

Introduction

The polycomb group (PcG) proteins play a central role in the maintenance of a repressive state of gene expression. Recent findings demonstrate that PcG components are organized into nuclear condensates, contributing to the reshaping of chromatin architecture in physiological and pathological conditions, thus affecting the nuclear mechanics. In this context, direct stochastic optical reconstruction microscopy (dSTORM) provides an effective tool to achieve a detailed characterization of PcG condensates by visualizing them at a nanometric level. Furthermore, by analyzing dSTORM datasets with cluster analysis algorithms, quantitative information can be yielded regarding protein numbers, grouping, and spatial organization.

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Principle Applications Procedure Materials Notes Related Products

Principle

Principles of stochastic optical reconstruction 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

Imaging in two or three dimensions, in multiple colors, and even in living cells
Applied in many areas of the life sciences, and provides very high resolution images for many different needs from neuroscience to subcellular science

Procedure

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

Materials

• Sample Type:
Polycomb group (PcG) proteins

Notes

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

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