Flow Cytometry for Filamentous Fungi

Abstract

Flow cytometry is a powerful high-throughput method, which enables a fast and multi-parameter analysis of single cells and particles. A plethora of different dyes for flow cytometry are available to label different parts of a cell in addition to in vivo markers like fluorescent proteins. Trichoderma species as well as other filamentous fungi show hyphal growth, which makes analysis in a flow cytometer difficult. Nevertheless, conidia can be readily analyzed in conventional flow cytometers. Many different applications can be envisaged. This protocol describes how conidia can be prepared for flow cytometry and the occurrence of genetic markers such as GFP can be measured. Furthermore, a guideline how to fix and stain cells is given.

Keywords: Conidia; FACS; Filamentous fungi; Fixation; Fluorescence; GFP; Sorting; Staining; Trichoderma reesei.