“Probe, Sample, and Instrument (PSI)”: The Hat-Trick for Fluorescence Live Cell Imaging

Ludovic Galas,Damien Schapman,Hitoshi Komuro,Arnaud Lehner,Patrice Lerouge,Stéphane Leleu,Alexis Lebon,Thibault Gallavardin,Xavier Franck,Magalie Bénard

doi:10.3390/chemosensors6030040

Abstract

Cell Imaging Platforms (CIPs) are research infrastructures offering support to a number of scientific projects including the choice of adapted fluorescent probes for live cell imaging. What to detect in what type of sample and for how long is a major issue with fluorescent probes and, for this, the “hat-trick” “Probe–Sample–Instrument” (PSI) has to be considered. We propose here to deal with key points usually discussed in CIPs including the properties of fluorescent organic probes, the modality of cell labeling, and the best equipment to obtain appropriate spectral, spatial, and temporal resolution. New strategies in organic synthesis and click chemistry for accessing probes with enhanced photophysical characteristics and targeting abilities will also be addressed. Finally, methods for image processing will be described to optimize exploitation of fluorescence signals.

Highlights

Since the 1960s, the discovery of fluorescent proteins [1,2] has boosted live cell imaging studies and, technological developments for microscopy including simultaneous resolution improvement [3,4,5,6] and detector sensitivity [7,8]
There is a large diversity of chemical and biomolecular strategies to construct one hand,probes there is a large diversity of chemical biomolecular strategies to construct newOn fluorescent with enhanced properties and and specificities
(B) Cell labeling with the naphtyl analogue of epicocconone through 2P microscopy

Summary

Introduction

Since the 1960s, the discovery of fluorescent proteins [1,2] has boosted live cell imaging studies and, technological developments for microscopy including simultaneous resolution improvement [3,4,5,6] and detector sensitivity [7,8]. In the early 2000s, European and French (Infrastructure en Biologie, Santé et Agronomie, IBiSA) policies were established to identify emerging cell imaging platforms (CIPs) in which equipment, human resources, and skills are mutualized to facilitate broad access to advanced technologies. By taking advantage of the diversity of projects and biological models studied on the Norman IBiSA. CIP, the so-called PRIMACEN (Plate-forme de Recherche en IMAgerie CEllulaire de Normandie), Chemosensors 2018, 6, 40; doi:10.3390/chemosensors6030040 www.mdpi.com/journal/chemosensors. 2018, 6, 40 biological studied on the Norman IBiSA CIP, the so-called PRIMACEN That is necessary to consider when choosing a fluorescent probe we propose here the concepttriad of the triad that is necessary to consider for live cell imaging (Figure 1). When choosing a fluorescent probe for live cell imaging (Figure 1). There is a large diversity of chemical and biomolecular strategies to construct one hand,probes there is a large diversity of chemical biomolecular strategies to construct newOn fluorescent with enhanced properties and and specificities

Objectives

Methods

Findings

Conclusion