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A Centrifuge Polarizing Microscope (CPM) That Enables the Visualization of Intracellular Structures Under High
Makoto Goda1,2, Akatsuki Kimura3,4,5
1Institute of Photonics Medicine, Hamamatsu University School of Medicine, Hamamatsu, Japan.
Abstract:
The mitotic spindle consists of aligned filaments of dynamic microtubules that faithfully segregate mitotic chromosomes. This view of the mitotic spindle was initially established by detecting weak birefringence of the aligned filaments, which was realized by developing polarized light microscopy (Inoué S, Chromosoma 5:487-500. https://doi.org/10.1007/bf01271498 , 1953). Inoué et al. developed a centrifuge polarizing microscope (CPM) (Inoué S et al., J Microsc 201:341-356. https://doi.org/10.1046/j.1365-2818.2001.00850.x , 2001; Inoué S et al., Centrifuge microscope capable of realizing polarized light observation (US Patent: US1907803A), 1999) to further understand the structural and physical basis of the functional positioning of centrosome-based structures (including the mitotic spindle) inside living cells. This chapter describes the procedure for observing biological specimens using CPM by focusing on Caenorhabditis elegans embryos.
Insights
Centrifuge polarizing microscopy (CPM) visualizes dynamic microtubules in the mitotic spindle, crucial for chromosome segregation. This method allows detailed observation of centrosome-based structures within living cells, exemplified using Caenorhabditis elegans embryos.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biophysics
Background:
- The mitotic spindle's structure and function are vital for accurate chromosome segregation during cell division.
- Polarized light microscopy has been instrumental in visualizing the aligned microtubule filaments of the mitotic spindle.
- Understanding the physical basis of centrosome-based structures in living cells requires advanced imaging techniques.
Purpose of the Study:
- To describe the procedure for observing biological specimens using Centrifuge Polarizing Microscopy (CPM).
- To highlight the utility of CPM in studying the structural and physical basis of centrosome-based structures.
- To demonstrate CPM application using Caenorhabditis elegans embryos.
Main Methods:
- Utilizing a Centrifuge Polarizing Microscope (CPM) for live-cell imaging.
- Observing the birefringence of dynamic microtubule filaments within the mitotic spindle.
- Applying the technique to Caenorhabditis elegans embryos as a model system.
Main Results:
- CPM enables visualization of the dynamic microtubules comprising the mitotic spindle.
- The method allows for the study of structural and physical properties of cellular components in vivo.
- Successful application of CPM to Caenorhabditis elegans embryos provides insights into spindle organization.
Conclusions:
- CPM is a powerful tool for investigating the mechanics of cell division and microtubule dynamics.
- The technique offers a unique approach to understanding the positioning of centrosome-based structures in live cells.
- This chapter provides a practical guide for researchers interested in applying CPM to their studies.
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