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Published on: April 7, 2017
Air-drying of cells enables visualization of antiparallel microtubule overlaps in the spindle midzone
Aya Ifuji1, Takahisa Kuga1, Yuji Nakayama1
1Department of Biochemistry & Molecular Biology, Kyoto Pharmaceutical University, Kyoto 607-8414, Japan.
Abstract:
Immunofluorescence staining is used extensively to examine various types of cellular events. However, even when an antibody can detect its epitopes in western blotting, it sometimes fails to detect its epitopes when used for immunofluorescence staining. One example is the antiparallel microtubule overlaps in the anaphase and telophase spindle midzone, which functions as a signaling scaffold for cleavage furrow specification. It has been believed that it cannot be visualized by immunofluorescence staining due to the highly dense structure of microtubule overlaps (Ifuji et al., 2017). Here, we show a simple method for visualization of antiparallel microtubule overlaps in the anaphase and telophase spindle midzone. •Air-drying cells before fixation enables visualization of antiparallel microtubule overlaps in the anaphase and telophase spindle midzone, which cannot be visualized by the conventional method.•Simple method that requires minimal usage of equipment.•Commonly used anti-tubulin antibodies can be used in this method.
Insights
A new air-drying method allows visualization of antiparallel microtubule overlaps in the spindle midzone, crucial for cell division. This simple technique overcomes limitations of conventional immunofluorescence staining.
Area of Science:
- Cell Biology
- Cytoskeletal Dynamics
Background:
- Immunofluorescence staining is vital for cellular event analysis.
- Conventional methods struggle to visualize dense structures like antiparallel microtubule overlaps in the spindle midzone.
Purpose of the Study:
- To develop a simple and effective method for visualizing antiparallel microtubule overlaps in the anaphase and telophase spindle midzone.
- To overcome the limitations of existing immunofluorescence techniques for dense microtubule structures.
Main Methods:
- A novel air-drying technique applied to cells before fixation.
- Utilizing commonly available anti-tubulin antibodies.
Main Results:
- Successfully visualized antiparallel microtubule overlaps in the anaphase and telophase spindle midzone.
- The air-drying method enables visualization where conventional methods failed due to dense microtubule structures.
Conclusions:
- Air-drying cells prior to fixation is a simple yet effective method for visualizing previously inaccessible antiparallel microtubule overlaps.
- This technique enhances the study of the spindle midzone's role in cleavage furrow specification.
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