Correlative single-cell X-ray tomography and X-ray fluorescence imaging

Authors: LIN, ZIHAN - Brookhaven National Laboratory; ZHANG, XIAO - Brookhaven National Laboratory; NANDI, PURBASHA - Brookhaven National Laboratory; LIN, YUEWEI - Brookhaven National Laboratory; WANG, LIGUO - Brookhaven National Laboratory; CHU, YONG - Brookhaven National Laboratory; Paape, Timothy - Tim; YANG, YANG - Brookhaven National Laboratory; XIAO, XIANGHUI - Brookhaven National Laboratory; LIU, QUN - Brookhaven National Laboratory

Submitted to: bioRxiv
Publication Type: Other
Publication Acceptance Date: 8/4/2023
Publication Date: 8/4/2023
Citation: Lin, Z., Zhang, X., Nandi, P., Lin, Y., Wang, L., Chu, Y., Paape, T.D., Yang, Y., Xiao, X., Liu, Q. 2023. Correlative single-cell X-ray tomography and X-ray fluorescence imaging. bioRxiv. https://doi.org/10.1101/2023.08.03.551868.
DOI: https://doi.org/10.1101/2023.08.03.551868

Interpretive Summary: X-ray tomography can provide images of internal structures using three-dimensional images. X-ray fluorescence is able to quantify metal ion distributions in animal or plant tissues, can also be applied to single cells such as bacteria. However X-rays can damage living cells and tissues. The method presented here can be used to compile images using a combination of x-ray fluorescence and x-ray tomography with significantly reduced damage to cells, at high resolution.

Technical Abstract: X-ray tomography and x-ray fluorescence imaging are two non-invasive imaging techniques to study cellular structures and chemical element distributions, respectively. However, correlative X-ray tomography and fluorescence imaging for the same cell has yet to be routinely realized due to challenges in sample preparation and X-ray radiation damage. Here we report an integrated experimental and computational workflow for achieving correlative multi-modality X-ray imaging of a single cell. The method consists of the preparation of radiation-resistant single-cell samples using live-cell imaging-assisted chemical fixation and freeze-drying procedures, targeting and labeling cells for correlative x-ray tomography and x-ray fluorescence measurement, and computational reconstruction of the correlative and multi-modality images. With X-ray tomography, cellular structures including the overall structure and intracellular organelles are visualized, while X-ray fluorescence imaging reveals the distribution of multiple chemical elements within the same cell. Our correlative method demonstrates the feasibility and broad applicability of using X-rays to understand cellular structures and the roles of multiple chemical elements and related proteins in signaling and other biological processes.