Applications of Light Sheet Microscopy in Microdevices
- 1Department of Bioengineering and Aerospace Engineering, Universidad Carlos III de Madrid, Spain
- 2Experimental Medicine and Surgery Unit, Instituto de Investigación Sanitaria Gregorio Marañón, Spain
- 3Experimental Medicine and Surgery Unit, Hospital General Universitario Gregorio Marañón, Spain
Light sheet fluorescence microscopy (LSFM) has been present in cell biology laboratories for quite some time, mainly as custom-made systems, with imaging applications ranging from single cells (in the micrometer scale) to small organisms (in the millimeter scale). Such microscopes distinguish themselves for having very low phototoxicity levels and high spatial and temporal resolution, properties that make them ideal for a large range of applications, including the study of dynamic processes in cell biology. In particular, cellular motion has proven to be essential in biological processes such as tumor metastasis and tissue development. Experimental setups make extensive use of microdevices (bioMEMS) that provide better control over the substrate environment than traditional cell culture experiments. For example, to mimic in vivo conditions, experiment biochemical dynamics, and trap, move or count cells. Microdevices provide higher degrees of empirical complexity but, so far, most have been designed to be imaged through wide-field or confocal microscopes. However, the properties of LSFM render it ideal for 3D characterization of active cells.
When working with microdevices, confocal microscopy is more widespread than LSFM even though it suffers from higher phototoxicity and slower acquisition speeds. Although it is sometimes possible to illuminate with a light sheet microdevices designed for confocal microscopes, these bioMEMS must be redesigned to exploit the full potential of LSFM. These properties are essential to image phenomena such as motion and traction, differentiation, and diffusion of molecules more frequently on a wider scale.
The use of microdevices for LSFM has extended beyond cell tracking studies into experiments regarding cytometry, spheroid cultures and lab-on-a-chip automation. Due to light sheet microscopy being in its early stages, a setup of these characteristics demands some degree of optical expertise; and designing three-dimensional microdevices requires facilities, ingenuity and experience in microfabrication. In this paper, we explore different approaches where light sheet microscopy can achieve single-cell and subcellular resolution within microdevices, and provide a few pointers on how these experiments may be improved.
Keywords: light-sheet fluorescence microscopy, SPIM, Microdevices, Microfluidics, Cellular Imaging
Received: 27 Sep 2018;
Accepted: 09 Jan 2019.
Edited by:Stéphane Pagès, Wyss Center for Bio and Neuroengineering, Switzerland
Reviewed by:Daniel A. Peterson, Rosalind Franklin University of Medicine and Science, United States
Giancarlo Ruocco, Istituto italiano di Tecnologia (IIT), Center for Life NanoScience, Italy
Copyright: © 2019 Albert-Smet, Marcos-Vidal, Vaquero, Desco, Muñoz-Barrutia and Ripoll. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Prof. Arrate Muñoz-Barrutia, Universidad Carlos III de Madrid, Department of Bioengineering and Aerospace Engineering, Getafe, 28903, Madrid, Spain, email@example.com
Prof. Jorge Ripoll, Universidad Carlos III de Madrid, Department of Bioengineering and Aerospace Engineering, Getafe, 28903, Madrid, Spain, firstname.lastname@example.org