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Autofluorescence imaging of human RPE cell granules by structured illumination microscopy (CAT#: STEM-MIT-0413-LJX)

Introduction

Retinal pigment epithelium, located between the choroid and photoreceptor extracellular segments, is a transport channel for ions, water, nutrients and metabolic end products between the lower retinal cavity and choroid blood vessels. Retinal pigment epithelium is involved in retinol circulation, phagocytosis of exfoliated photoreceptor extracellular segments to maintain photoreceptor cell excitability, and secretes a variety of growth factors to help maintain the structural integrity of choroidal vascular endothelial cells and photoreceptor cells.




Principle

The structured illumination microscopy (SIM) applies a pattern lighting field (different from the traditional wide-field lighting) to the samples to improve the spatial resolution of the optical microscope and has advantages for the observation of living cells. In this method, the spatial frequency of the illumination pattern is mixed with the spatial frequency of the sample feature, converting the high frequency feature into a lower frequency detectable by the microscope. The periodic lighting pattern (Moire fringes, Moire fringes) is generated by the interference of multiple light sources in the axial (Z), lateral (X-Y) or both directions, and the high-resolution image is reconstructed based on the acquisition of multiple illumination images in different phases and directions. Since the illumination mode itself is also limited by optical diffraction, SIM can only double the spatial resolution by combining two information sources with limited diffraction, achieving resolutions of 100 nm and 300 nm in the X-Y and Z-axis directions, respectively.

Applications

Applied to the research of cell physiology, cell dynamics and other subcellular level

Procedure

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

Materials

• Sample Type:
Human RPE cell granules

Notes

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