A novel correlative 3D microscopy platform for studying cells in their native state

Authors: Ilias Kounatidis (Diamond Light Source)
Co-authored by industrial partner: No

Type: Diamond Annual Review Highlight

State: Published (Approved)
Published: July 2021
Diamond Proposal Number(s): 21046 , 18314

Abstract: Researchers have developed a new technique for studying cells in their native state. The goal was to obtain high-quality imaging data from cells without the need for sectioning or chemical fixation. The new method avoids any treatment that would disturb cell structure, so that no artefacts (errors) are introduced into the images. To demonstrate this novel correlative microscopy platform’s effectiveness, the team studied the early stages of cell infection by reoviruses. Although the specific viruses have been studied extensively, there is a debate regarding the method of infection. This research focused on the way that the virus escapes from vesicles, a required step for replication. At beamline B24, using a correlative imaging approach by combining soft X-ray tomography with super resolution microscopy in cryogenic conditions, the team tracked the infection mechanism. The results revealed that the virus had already escaped from the host vesicles two hours after infection, with the vesicles preserving their circular shape, suggesting a gentle, pore-based exit mechanism for the virus. Reoviruses are valuable tools that could be engineered to express proteins and have the potential to be used in vaccines. Knowing the infection mechanism will facilitate their handling and manipulation for biomedical purposes. The new imaging platform has also been used to validate anticancer compounds, study cell structure during development and investigate clearance of human pathogenic microorganism by immune host cells. The work is the outcome of a collective effort between Diamond Light Source and research groups and facilities across Europe, including the University of Oxford, Heidelberg University Hospital, the Université de Nantes and CryoCapCell.

Journal Keywords: Correlative imaging; Structured illumination microscopy; X-ray tomography; Reovirus biology; Correlative light/X-ray tomography (CLXT)

Subject Areas: Technique Development, Biology and Bio-materials

Instruments: B24-Cryo Soft X-ray Tomography

Added On: 12/11/2021 10:54

Discipline Tags:

Pathogens Infectious Diseases Health & Wellbeing Technique Development - Life Sciences & Biotech Life Sciences & Biotech

Technical Tags:

Microscopy X-ray Microscopy Correlative Light X-ray Microscopy (CLXM)