Analyzing self-assembled spindle dynamics in fission yeast meiosis using in vivo fluorescence imaging

Alberto Pineda-Santaella; Rebeca Martín-García; Alfonso Fernández-Álvarez

doi:10.1016/j.xpro.2023.102655

Analyzing self-assembled spindle dynamics in fission yeast meiosis using in vivo fluorescence imaging

STAR Protoc. 2023 Dec 15;4(4):102655. doi: 10.1016/j.xpro.2023.102655. Epub 2023 Oct 20.

Authors

Alberto Pineda-Santaella¹, Rebeca Martín-García¹, Alfonso Fernández-Álvarez²

Affiliations

¹ Instituto de Biología Funcional y Genómica (IBFG), Consejo Superior de Investigaciones Científicas (CSIC) and Universidad de Salamanca, Calle Zacarías González 2, 37007 Salamanca, Spain.
² Instituto de Biología Funcional y Genómica (IBFG), Consejo Superior de Investigaciones Científicas (CSIC) and Universidad de Salamanca, Calle Zacarías González 2, 37007 Salamanca, Spain. Electronic address: alfonso.fernandez.alvarez@csic.es.

Abstract

Chromosome segregation in female meiosis in many metazoans is mediated by acentrosomal spindles. The analysis of the dynamics of self-assembled spindles is a challenge due to the low availability of oocytes. Here, we present a protocol for analyzing self-assembled spindle dynamics in fission yeast meiosis using in vivo fluorescence imaging. We describe steps for starter culture preparation, meiosis induction, and sample preparation. We then detail procedures for acquisition and analysis of images of self-assembled spindles. For complete details on the use and execution of this protocol, please refer to Pineda-Santaella and Fernández-Álvarez (2019)¹ and Pineda-Santaella et al. (2021).².

Keywords: Cell Biology; Microscopy; Model Organisms.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Female
Humans
Meiosis
Microtubules
Optical Imaging
Schizosaccharomyces*
Spindle Apparatus*