Imaging of peroxisomes by stochastic optical reconstruction microscopy (CAT#: STEM-MIT-0372-LJX)

Introduction

Peroxisomes are central metabolic organelles whose maturation and function depend on efficient and accurate targeting of peroxisomal membrane proteins (PMPs). Ultrastructural imaging of the PMPs is a quite difficult task as it requires high spatial and temporal resolution. Further, the spatial resolution of conventional light microscopy is limited due to the diffraction of light. However, recent methodological developments in super resolution microscopy showed us to access the nanoscale regimes spatially allowing to elucidate the membrane structures of cell organelles.

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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:
Peroxisomes

Notes

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

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