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Imaging in Drosophila Brain Slices by Stimulated Emission Depletion Microscopy (CAT#: STEM-MIT-0364-LJX)

Introduction

Super-resolution microscopy is a very powerful tool to investigate fine cellular structures and molecular arrangements in biological systems. For instance, stimulated emission depletion (STED) microscopy has been successfully used in recent years to investigate the arrangement and colocalization of different protein species in cells in culture and on the surface of specimens. However, because of its extreme sensitivity to light scattering, super-resolution imaging deep inside tissues remains a challenge. Here, we describe the preparation of thin slices from the fruit fly (Drosophila melanogaster) brain, subsequent immunolabeling and imaging with STED microscopy. This protocol allowed us to image small dendritic branches from neurons located deep in the fly brain with improved resolution compared with conventional light microscopy.




Principle

Stimulated emission depletion (STED) microscopy uses two light sources. One source emits light that excites the fluorophores, and the other emits a ring laser of different wavelengths, which is used to suppress fluorescence.

Applications

Imaging of the intensity distribution of the fluorescent sample
Imaging of living samples
Measuring of the fluorescence lifetime and fluorescence correlation spectrum of the fluorescent samples
Used in the fields of biology, medicine and materials science

Procedure

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

Materials

• Sample Type:
Drosophila Brain Slices

Notes

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