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Imaging of mouse glutamatergic synaptosomes by cryo-laser scanning confocal microscope technology (CAT#: STEM-MIT-0090-LJX)

Introduction

Ionotropic glutamate receptors (iGluRs) at postsynaptic terminals mediate the majority of fast excitatory neurotransmission in response to release of glutamate from the presynaptic terminal. Obtaining structural information on the molecular organization of iGluRs in their native environment, along with other signaling and scaffolding proteins in the postsynaptic density (PSD), and associated proteins on the presynaptic terminal, would enhance understanding of the molecular basis for excitatory synaptic transmission in normal and in disease states. Cryo-electron tomography (ET) studies of synaptosomes is one attractive vehicle by which to study iGluR-containing excitatory synapses.<br />Cryo-laser scanning confocal microscope (cryo-LSM) can be used to rapidly detect the fluorescence labels at the presynaptic and postsynaptic terminals of glutamate synaptosomes.




Principle

Laser scanning confocal microscope is a high-tech microscope. It is based on fluorescence microscope imaging and equipped with a laser scanning device, which uses ultraviolet or visible light to excite the fluorescence probe, thereby obtaining fluorescence images of the internal microstructure of cells or tissues.
The laser beam is used as the light source in the laser scanning confocal microscope. The laser beam passes through the illuminating pinhole and is reflected to the objective lens through the spectroscope. The laser beam is focused on the sample, and every point on the focal plane of the specimen is scanned. If there is a fluorescent substance that can be excited in the tissue sample, the fluorescence emitted after excitation is directly reversed back to the spectroscope through the original incident light path, and is first focused when passing through the detection pinhole. The focused light is detected and collected by the photomultiplier tube (PMT), and the signal is sent to the computer, and the image is displayed on the computer monitor after processing.

Applications

Imaging and analysis in the fields of morphology, molecular cell biology, neuroscience, pharmacology, genetics

Procedure

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

Materials

• Sample Type:
Mouse glutamatergic synaptosomes

Notes

Operate in strict accordance with the operating procedures, and shall not arbitrarily change the operating procedures
In the starting sequence of the switch and in the scanning process, try to do fast and orderly, to protect the laser