Abstract
X-ray imaging is a complementary method to electron and fluorescence microscopy for studying biological cells. In particular, scanning small-angle X-ray scattering provides overview images of whole cells in real space as well as local, high-resolution reciprocal space information, rendering it suitable to investigate subcellular nanostructures in unsliced cells. One persisting challenge in cell studies is achieving high throughput in reasonable times. To this end, afastscanning mode is used to image hundreds of cells in a single scan. A way ofdealing with the vast amount of data thus collected is suggested, including asegmentation procedure and three complementary kinds of analysis, i.e. characterization of the cell population as a whole, of single cells and of different parts of the same cell. The results show that short exposure times, which enable faster scans and reduce radiation damage, still yield information in agreement with longer exposure times.
Highlights
Imaging biological cells with a spatial resolution sufficient for identifying subcellular structures is a very challenging task, currently tackled mainly by three kinds of probes: electrons, visible-light fluorescence and X-rays
We demonstrate that fast scanning small-angle X-ray scattering (SAXS) experiments on a large number of mammalian cells are possible, thanks to the synchronization of the continuous movement of the sample stage with the data acquisition (Nicolas et al, 2017)
We achieve high throughput, as we examine roughly 800 cells, storing 8 895 234 scattering patterns in about 7 h
Summary
Imaging biological cells with a spatial resolution sufficient for identifying subcellular structures is a very challenging task, currently tackled mainly by three kinds of probes: electrons, visible-light fluorescence and X-rays. Electron microscopy (Koster & Klumperman, 2003; de Jonge et al, 2009) yields the best spatial resolution, resolving details down to the subnanometer range. It requires extensive sample preparation, typically including slicing and staining of the sample. Thanks to super-resolution techniques (Hell, 2007), fluorescence microscopy is widely used in labeled, intact cells (Fernandez-Suarez & Ting, 2008; Huang et al, 2010) and can resolve details on the order of tens of nanometers. Scanning small-angle X-ray scattering (SAXS) (Fratzl et al, 1997) is used on unsliced, unstained samples to obtain both real and reciprocal space information.
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