On Some Current Challenges in High-Resolution Optical Bioimaging

Abstract

In this Perspective we propose our current point of view and a suggestive roadmap on the field of high-resolution optical microscopy dedicated to bioimaging. Motivated by biological applications, researchers have indeed devised an impressive amount of strategies to address the diverse constraints of imaging and studying biological matter down to the molecular scale, making this interdisciplinary research field a vibrant forum for creativity. Throughout the discussion, we highlight several striking recent successes in this quest. We also identify some next challenges still ahead to apprehend biological questions in increasingly complex living organisms for integrative studies in a minimally invasive manner.

Figure 1

Super-resolution fluorescence microscopy of nuclear pore complexes with different modalities: STED microscopy (a), compared with confocal microscopy; STORM microscopy (b); DNA PAINT microscopy (c, d). Comparison to electron microscopy using negative staining. Republished with permission of Company of Biologist Ltd. from ref (22). Copyright 2012 The Company of Biologists, permission conveyed through Copyright Clearance Center, Inc.; Adapted with permission from ref (23). Copyright 2019 John Wiley and Sons, https://creativecommons.org/licenses/by/4.0/; Reprinted with permission from ref (24). Copyright 2013 Elsevier.

Figure 2

Living cell (mouse embryonic fibroblast) observed with (a) fluorescence imaging (mitochondrial labeling), (b) transmission intensity, (c) quantitative phase, (d) phase contrast (simulated from (c)), and (e) differential interference contrast (simulated from (c)). The setup used is described in ref (37).

Figure 3

Near-infrared bioimaging with nanoparticles. (a, b) The different near-infrared (NIR) region biological transparency windows. (c) NIR in vivo imaging of colon cancer mouse model tumors injected with rare-earth nanoparticles. (c, d) NIRII single particle tracking of single wall carbon nanotubes (SWCNTs) in complex environments (d: schematics) allow the measurement of super-resolved maps of the extracellular space in live brain slices. Reprinted with permission from (52). Copyright 2009 Springer Customer Service Center GmbH: Springer Nature; Adapted with permission from ref (53). Copyright 2021 Springer Nature, https://creativecommons.org/licenses/by/4.0/; Reprinted with permission from ref (56). Copyright 2019 Springer Customer Service Center GmbH: Springer Nature; Reprinted with permission from ref (60). Copyright 2017 Springer Customer Service Center GmbH: Springer Nature.