Analyzing maize meiotic chromosomes by super-resolution structured illumination microscopy (CAT#: STEM-MIT-0412-LJX)

Introduction

The success of meiosis depends on intricate coordination of a series of unique cellular processes to ensure proper chromosome segregation. Many proteins involved in these cellular events are directly or indirectly associated with chromosomes, especially those required for homologous recombination. These meiotic processes have been explored extensively by conventional light microscopy. However, many features of interest, such as chromatin organization, recombination nodules, or the synaptonemal complex are beyond the resolution of conventional wide-field microscopy. Moreover, in most sample preparation techniques for light microscopy, meiotic cells are squashed, which destroys the spatial organization of the nucleus. The service provides a protocol to analyze maize meiotic chromosomes by three-dimensional structured illumination microscopy (3D-SIM), a recently developed high-resolution microscopy technique. This protocol can be used to examine protein localizations at a high resolution level by immunofluorescence.

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Principle Applications Procedure Materials Notes Related Products

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:
Maize meiotic chromosomes

Notes

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

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