Ivanov Ivan E, Yeh Li-Hao, Perez-Bermejo Juan A, Byrum Janie R, Kim James Y S, Leonetti Manuel D, Mehta Shalin B
Chan Zuckerberg Biohub, 499 Illinois St, San Francisco, CA 94158, USA.
Gladstone Institutes, 1650 Owens St, San Francisco, CA 94158, USA.
Biomed Opt Express. 2022 Apr 27;13(5):3102-3119. doi: 10.1364/BOE.455770. eCollection 2022 May 1.
The spatial and angular organization of biological macromolecules is a key determinant, as well as informative readout, of their function. Correlative imaging of the dynamic spatio-angular architecture of cells and organelles is valuable, but remains challenging with current methods. Correlative imaging of spatio-angular dynamics requires fast polarization-, depth-, and wavelength-diverse measurement of intrinsic optical properties and fluorescent labels. We report a multimodal instant polarization microscope (miPolScope) that combines a broadband polarization-resolved detector, automation, and reconstruction algorithms to enable label-free imaging of phase, retardance, and orientation, multiplexed with fluorescence imaging of concentration, anisotropy, and orientation of molecules at diffraction-limited resolution and high speed. miPolScope enabled multimodal imaging of myofibril architecture and contractile activity of beating cardiomyocytes, cell and organelle architecture of live HEK293T and U2OS cells, and density and anisotropy of white and grey matter of mouse brain tissue across the visible spectrum. We anticipate these developments in joint quantitative imaging of density and anisotropy to enable new studies in tissue pathology, mechanobiology, and imaging-based screens.
生物大分子的空间和角度组织是其功能的关键决定因素及信息读出方式。对细胞和细胞器的动态时空角度结构进行相关成像很有价值，但用当前方法仍具有挑战性。时空角度动力学的相关成像需要对固有光学特性和荧光标记进行快速的偏振、深度和波长多样的测量。我们报告了一种多模态即时偏振显微镜（miPolScope），它结合了宽带偏振分辨探测器、自动化和重建算法，能够在衍射极限分辨率和高速下，对相位、延迟和取向进行无标记成像，并与分子浓度、各向异性和取向的荧光成像进行多路复用。miPolScope实现了对肌原纤维结构以及跳动心肌细胞收缩活动、活HEK293T和U2OS细胞的细胞和细胞器结构，以及小鼠脑组织白质和灰质在可见光谱范围内的密度和各向异性的多模态成像。我们预计这些在密度和各向异性联合定量成像方面的进展将推动组织病理学、力学生物学和基于成像的筛选等新研究。
