Air-drying of cells enables visualization of antiparallel microtubule overlaps in the spindle midzone

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. Method name: Air-drying method, Keywords: Anaphase, Antiparallel microtubule overlap, Immunofluorescence staining, Microtubule, Midzone, Mitosis, Telophase