Tosheva Kalina L, Yuan Yue, Matos Pereira Pedro, Culley Siân, Henriques Ricardo
MRC Laboratory for Molecular Cell Biology, University College London, London, United Kingdom.
ITQB-NOVA, Oeiras, Portugal.
J Phys D Appl Phys. 2020 Apr 15;53(16):163001. doi: 10.1088/1361-6463/ab6b95. Epub 2020 Feb 14.
Super-resolution microscopy (SRM) enables non-invasive, molecule-specific imaging of the internal structure and dynamics of cells with sub-diffraction limit spatial resolution. One of its major limitations is the requirement for high-intensity illumination, generating considerable cellular phototoxicity. This factor considerably limits the capacity for live-cell observations, particularly for extended periods of time. Here, we give an overview of new developments in hardware, software and probe chemistry aiming to reduce phototoxicity. Additionally, we discuss how the choice of biological model and sample environment impacts the capacity for live-cell observations.
超分辨率显微镜(SRM)能够以亚衍射极限空间分辨率对细胞内部结构和动态进行非侵入性、分子特异性成像。其主要局限性之一是需要高强度照明,这会产生相当大的细胞光毒性。这一因素极大地限制了活细胞观察的能力,尤其是长时间观察的能力。在此,我们概述了旨在降低光毒性的硬件、软件和探针化学方面的新进展。此外,我们还讨论了生物模型和样品环境的选择如何影响活细胞观察的能力。