Publication
Article Metrics
Citations
Online attention
Cryo-structured illumination microscopic data collection from cryogenically preserved cells
Authors:
Nina
Vyas
(Diamond Light Source)
,
Nina
Perry
(Diamond Light Source)
,
Chidinma A.
Okolo
(Diamond Light Source)
,
Ilias
Kounatidis
(Diamond Light Source)
,
Thomas M.
Fish
(Diamond Light Source)
,
Kamal L.
Nahas
(Dimond Light Source; University of Cambridge)
,
Archana
Jadhav
(Diamond Light Source)
,
Mohamed A.
Koronfel
(Diamond Light Source)
,
Johannes
Groen
(ALBA Synchrotron)
,
Eva
Pereiro
(ALBA Synchrotron)
,
Ian M.
Dobbie
(University of Oxford)
,
Maria
Harkiolaki
(Diamond Light Source)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Journal Of Visualized Experiments
State:
Published (Approved)
Published:
May 2021
Diamond Proposal Number(s):
25512

Abstract: Three-dimensional (3D) structured illumination microscopy (SIM) allows imaging of fluorescently labelled cellular structures at higher resolution than conventional fluorescence microscopy. This super-resolution (SR) technique enables visualization of molecular processes in whole cells and has the potential to be used in conjunction with electron microscopy and X-ray tomography to correlate structural and functional information. A SIM microscope for cryogenically preserved samples (cryoSIM) has recently been commissioned at the correlative cryo-imaging beamline B24 at the UK synchrotron. It was designed specifically for 3D imaging of biological samples at cryogenic temperatures in a manner compatible with subsequent imaging of the same samples by X-ray microscopy methods such as cryo-soft X-ray tomography. This video article provides detailed methods and protocols for successful imaging using the cryoSIM. In addition to instructions on the operation of the cryoSIM microscope, recommendations have been included regarding the choice of samples, fluorophores, and parameter settings. The protocol is demonstrated in U2OS cell samples whose mitochondria and tubulin have been fluorescently labelled.
Subject Areas:
Biology and Bio-materials,
Technique Development
Instruments:
B24-Cryo Soft X-ray Tomography
Added On:
21/06/2021 09:19
Discipline Tags:
Technique Development - Life Sciences & Biotech
Structural biology
Life Sciences & Biotech
Technical Tags:
Microscopy
X-ray Microscopy
Cryo X-ray Microscopy