Live cell microscopy: From image to insight

Author:

Cuny Andreas P.12ORCID,Schlottmann Fabian P.3ORCID,Ewald Jennifer C.3ORCID,Pelet Serge4ORCID,Schmoller Kurt M.56ORCID

Affiliation:

1. Department of Biosystems Science and Engineering, ETH Zurich, 4058 Basel, Switzerland

2. Swiss Institute of Bioinformatics, 4058 Basel, Switzerland

3. Interfaculty Institute of Cell Biology, University of Tuebingen, 72076 Tuebingen, Germany

4. Department of Fundamental Microbiology, University of Lausanne, 1015 Lausanne, Switzerland

5. Institute of Functional Epigenetics, Helmholtz Zentrum München, 85764 Neuherberg, Germany

6. German Center for Diabetes (DZD), 85764 Neuherberg, Germany

Abstract

Live-cell microscopy is a powerful tool that can reveal cellular behavior as well as the underlying molecular processes. A key advantage of microscopy is that by visualizing biological processes, it can provide direct insights. Nevertheless, live-cell imaging can be technically challenging and prone to artifacts. For a successful experiment, many careful decisions are required at all steps from hardware selection to downstream image analysis. Facing these questions can be particularly intimidating due to the requirement for expertise in multiple disciplines, ranging from optics, biophysics, and programming to cell biology. In this review, we aim to summarize the key points that need to be considered when setting up and analyzing a live-cell imaging experiment. While we put a particular focus on yeast, many of the concepts discussed are applicable also to other organisms. In addition, we discuss reporting and data sharing strategies that we think are critical to improve reproducibility in the field.

Funder

German Research Foundation

Human Frontiers Science Program

Helmholtz Gesellschaft

Swiss National Science Foundation

Publisher

AIP Publishing

Subject

General Medicine

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