Imaging of In Vitro and In Vivo Neurons in Drosophila by Stochastic Optical Reconstruction Microscopy (CAT#: STEM-MIT-0380-LJX)

Introduction

The Drosophila melanogaster brain comprises different neuronal cell types that interconnect with precise patterns of synaptic connections. These patterns are essential for the normal function of the brain. To understand the connectivity patterns requires characterizing them at single-cell resolution, for which a fluorescence microscope becomes an indispensable tool. Additionally, because the neurons connect at the nanoscale, the investigation often demands super-resolution microscopy. Here, we adopt one super-resolution microscopy technique, called stochastic optical reconstruction microscopy (STORM), improving the lateral and axial resolution to ∼20 nm.

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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:
Neurons in Drosophila

Notes

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

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