Automatic Counting of Intra-Cellular Ribonucleo-Protein Aggregates in Saccharomyces cerevisiae Using a Textural Approach

Microsc Microanal. 2019 Feb;25(1):164-179. doi: 10.1017/S1431927619000084. Epub 2019 Feb 13.

Abstract

In the context of microbiology, recent studies show the importance of ribonucleo-protein aggregates (RNPs) for the understanding of mechanisms involved in cell responses to specific environmental conditions. The assembly and disassembly of aggregates is a dynamic process, the characterization of the stage of their evolution can be performed by the evaluation of their number. The aim of this study is to propose a method to automatically determine the count of RNPs. We show that the determination of a precise count is an issue by itself and hence, we propose three textural approaches: a classical point of view using Haralick features, a frequency point of view with generalized Fourier descriptors, and a structural point of view with Zernike moment descriptors (ZMD). These parameters are then used as inputs for a supervised classification in order to determine the most relevant. An experiment using a specific Saccharomyces cerevisiae strain presenting a fusion between a protein found in RNPs (PAB1) and the green fluorescent protein was performed to benchmark this approach. The fluorescence was observed with two-photon fluorescence microscopy. Results show that the textural approach, by mixing ZMD with Haralick features, allows for the characterization of the number of RNPs.

Keywords: Generalized Fourier descriptors; Haralick; Zernike; ribonucleo-protein aggregates; sub-cellular structures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytoplasm*
  • Cytoplasmic Granules / metabolism
  • Green Fluorescent Proteins
  • Microscopy, Fluorescence / methods*
  • Models, Biological
  • Poly(A)-Binding Proteins / isolation & purification
  • Protein Aggregates*
  • Ribonucleoproteins / isolation & purification*
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / isolation & purification

Substances

  • Poly(A)-Binding Proteins
  • Protein Aggregates
  • Ribonucleoproteins
  • Saccharomyces cerevisiae Proteins
  • pab1 protein, S cerevisiae
  • Green Fluorescent Proteins