Biology Reference
In-Depth Information
CHAPTER
Fission Yeast
24
Judite Costa
*
,{
, Chuanhai Fu
{
, Viktoriya Syrovatkina
*
and Phong T. Tran
*
,{
Imaging Individual Spindle
Microtubule Dynamics in
*
Cell and Developmental Biology, University of Pennsylvania, Philadelphia, Pennsylvania USA
{
Cell Biology, Institut Curie, UMR 144 CNRS, Paris, France
{
Biochemistry, University of Hong Kong, Pokfulam, Hong Kong
CHAPTER OUTLINE
Introduction ............................................................................................................ 386
24.1 Methods ........................................................................................................ 387
24.1.1 PDMS Slide Spacer ................................................................... 387
24.1.2 Slide Assembly ......................................................................... 388
24.1.3 Live-cell Imaging....................................................................... 390
24.1.4 Data Analysis ............................................................................ 391
Conclusion ............................................................................................................. 393
Acknowledgments ................................................................................................... 393
References ............................................................................................................. 394
Abstract
Microtubules exhibit dynamic instability, stochastically switching between infre-
quent phases of growth and shrinkage. In the cell, microtubule dynamic instability
is further modulated by microtubule-associated proteins and motors, which are spe-
cifically tuned to cell cycle stages. For example, mitotic microtubules are more dy-
namic than interphase microtubules. The different parameters of microtubule
dynamics can be measured from length versus time data, which are generally
obtained from time-lapse acquisition using the optical microscope. The typical max-
imum resolution of the optical microscope is
300 nm. This scale represents
a challenge for imaging fission yeast microtubule dynamics specifically during early
mitosis, where the bipolar mitotic spindle contains many short dynamic microtubules
of
l
/2 or
1-
m
m scale. Here, we present a novel method to image short fission yeast mitotic
