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Short- and long-term observations of mobile and immobile endoplasmic reticulum in onion bulb epidermis cells by a confocal laser scanning microscope technology (CAT#: STEM-MIT-0131-LJX)

Introduction

The endoplasmic reticulum (ER) of onion bulb scale epidermis cells consists of long, tubular strands lying deep in the cytoplasm which move quickly and a less mobile peripheral network of tubules and cisternae that change in position, shape and size but that also have immobile, fixed, sites (IFSs). IFSs occur in junctions, at vertexes and at blind endings of tubules as well as at the edges and the surface of cisternae. They are regularly arranged in helicoidal rows and may be knot- or ring-like in structure. They become enlarged by treatment with oryzalin but not with colchicine. They persist for long times (for more than 30 min); together with pulling forces, the surface tension and other factors, they determine the configuration and motion of the peripheral network. New polygons of the network are mainly formed by the development of new tubules that become joined with other parts of the network. Polygons disappear by contraction and fusion of tubules.




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:
Onion bulb epidermis cells

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
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