Applications of Soft X-ray Tomography for the direct observation of native cellular events

DOI: 10.1093/micmic/ozad067.586

Authors: Chidinma Okolo (Diamond Light Source) , Archana Jadhav (Diamond Light Source) , Thomas Fish (Diamond Light Source) , Kamal Nahas (Dimond Light Source, Diamond Light Source) , Amy Watts (Diamond Light Source) , Maria Harkiolaki (Diamond Light Source)
Co-authored by industrial partner: No

Type: Conference Paper
Conference: Microscopy and Microanalysis 2023
Peer Reviewed: No

State: Published (Approved)
Published: August 2023

Abstract: Cryo-Soft X-ray Tomography (cryoSXT) is an emerging 3D imaging tool for the investigation of the cellular world under near physiological states in 3D and to a resolution of tens of nanometers. Imaging can take place over substantial fields of views (10–16 μ which can be sequentially stitched to form 3D mosaics of hundreds of microns in samples that only need to be snap frozen and therefore remain representative of the structures within fully hydrated living cells [1]. As cryoSXT exploits the natural absorption of a specified range soft X-rays by carbonaceous biological matter there is no requirement for chemical staining or otherwise processing of samples minimising therefore the risk for artefact formation [2]. As the methodology has matured over the past few years, the need has arisen for well mapped processes in sample preparation, data collection and data analyses as well as correlation with other imaging methods to allow us to harness the power of more than one technique for any one research endeavor. Concurrently the need to standardize and quantify the quality and content of cryoSXT data collected at different locations and over different experiments necessitates the design of archiving and reporting protocols that will democratise access to the accumulated data for the benefit of the broader scientific community and researchers that are not necessarily technique experts. Here, the theory and current implementations of cryoSXT will be presented alongside the latest developments in the field and the research that has been enabled through its use. Data from the UK synchrotron beamline B24 will be used to exemplify applications including correlative 3D imaging (Figure 1) and method automation (Figure 2) that highlights the pressing need for cryoSXT availability in biomedical research. New, user-friendly decision matrices developed at B24 that aim to both simplify and standardize the use of this method will be also presented and the landscape which lies ahead will be explored.

Subject Areas: Technique Development, Biology and Bio-materials


Instruments: B24-Cryo Soft X-ray Tomography

Added On: 23/08/2023 14:26

Discipline Tags:

Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags:

Imaging Tomography Cryo-soft X-ray Tomography (Cryo-SXT)