Estimating Microtubule Distributions from 2D Immunofluorescence Microscopy Images Reveals Differences among Human Cultured Cell Lines

Jonathan A Coles,Jieyue Li,Aabid Shariff,Emma Lundberg,Robert F Murphy,Gustavo K Rohde,Mikaela Wiking

doi:10.1371/journal.pone.0050292

Abstract

Microtubules are filamentous structures that are involved in several important cellular processes, including cell division, cellular structure and mechanics, and intracellular transportation. Little is known about potential differences in microtubule distributions within and across cell lines. Here we describe a method to estimate information pertaining to 3D microtubule distributions from 2D fluorescence images. Our method allows for quantitative comparisons of microtubule distribution parameters (number of microtubules, mean length) between different cell lines. Among eleven cell lines compared, some showed differences that could be accounted for by differences in the total amount of tubulin per cell while others showed statistically significant differences in the balance between number and length of microtubules. We also observed that some cell lines that visually appear different in their microtubule distributions are quite similar when the model parameters are considered. The method is expected to be generally useful for comparing microtubule distributions between cell lines and for a given cell line after various perturbations. The results are also expected to enable analysis of the differences in gene expression underlying the observed differences in microtubule distributions among cell types.

Highlights

Microtubules play an indispensable role in subcellular processes such as cell movement, cell division and intracellular transportation
While limited information is available about variation in microtubule distributions [1,2], information on those distributions in intact cells for different cell lines has not been readily available
Since the images we analyze in this paper are only 2D slices, we developed an approach to estimate an approximate 3D shape of a cell and nucleus from a 2D slice

Summary

Introduction

Microtubules play an indispensable role in subcellular processes such as cell movement, cell division and intracellular transportation. These processes are known to play a role in other biological phenomena such as wound healing, and cancer metastasis. Extracting information about the organization of microtubules in different cell lines could potentially shed light on the roles of microtubule associated proteins in that organization. Most microtubule studies have focused on dynamics and interactions with drugs and microtubule associated proteins [3,4,5,6]. We believe that the ability to obtain reliable estimates of the overall organization of microtubules in whole cells could allow quantification of their dependency on different pertubagens, drugs, mechanical stimuli, etc

Methods

Results

Conclusion