Study of lamellar bodies and pulmonary surfactant protein A in isolated alveolar type II cells by confocal laser scanning immunofluorescence microscopy technology (CAT#: STEM-MIT-0118-LJX)

Introduction

Rat type II alveolar epithelial cells were isolated from lung tissue. Alveoli are hemispherical vesicles made of a single layer of epithelial cells. The bronchi in the lung are branched many times into numerous bronchioles, whose ends are enlarged into sacs surrounded by many small protruding sacs called alveoli. Small alveolar cells, also known as type I alveolar cells, do not proliferate. Large alveolar cells, also known as type II alveolar cells, secrete a surface active substance to reduce alveolar surface tension.
Lung surfactant associated protein A is the first protein found and strongly expressed in alveolar type II epithelial cells with the most abundant signal. It is one of the important components of lung surfactant, and its function, synthesis and secretion regulation are complex, which is of great clinical significance.

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Principle Applications Procedure Materials Notes Related Products

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:
Type II cells isolated from rat lungs

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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