Reports

Biostruct-X Workshop on Cryo X-ray Imaging for Biological Applications
ALBA, 22nd March 2013
The aim of the workshop was bringing together experts and users of all levels to exchange expertise and knowledge to strengthen the technique potentials and help disseminating its possibilities. Along these lines, two sessions were organized. The morning session was dedicated mainly to sample preparation and correlative microscopy (fluorescence microscopy/electron microscopy/X-ray microscopy) while the afternoon session was dedicated to data processing.
Procedures on how to prepare successfully a sample to be able to collect good data were explained in detail. Sample preparation requires depositing or growing the cells in appropriate microscopy grids and vitrification using plunge-freezing or high pressure freezing. To increase the probability of having a successful sample preparation for the X-ray imaging experiment, samples have to be checked before and after vitrification. An overview on different research topics was given by scientists who have used X-ray imaging correlatively to fluorescence and electron microscopy to answer specific scientific questions.
The data processing session started with an overview on X-ray image formation where the problem of the depth of focus was introduced as well as a strategy to deal with it. Then a detailed description of the different reconstruction software packages that are available from the electron microscopy community was presented by the creators of these packages. After all talks and during the breaks, very fruitful and lively discussions took place between the users and experts.
There were 46 registered participants coming from France (3), UK (10), Germany (5), Israel (2), USA (5), Taiwan (4) and Spain (17). We invited 3 experts on software reconstruction from the Electron Microscopy community that have created packages that are now used as well in X-ray image processing with the idea of comparing them and make them known to the community: David Mastronarde (Colorado University, USA), creator of IMOD package, Bernard Heymann (LSBR/NIAMS/NIH, USA), creator of BSoft, Sergio Marco (Institut Curie, France), creator of TomoJ. These experts are keen to work on further developments of their packages to incorporate the special needs of X-ray microscopy like the depth of focus problem.
The feedback from the participants clearly shows that starting users are requesting a hands-on session on data reconstruction using a specific software package to learn how to deal with their data when they have little experience in tomographic reconstruction; while expert users would have preferred a 2-days meeting to increase the discussion between participants. More specific meetings, such as a hands-on on data reconstruction, could follow this one to help starting users. A yearly meeting such as the one held at ALBA should be contemplated during all the length of the Biostruct-X project to strengthen the use of X-ray imaging in the structural biologist community.
A mailing list named “3dtomox” (3dtomox@cnb.csic.es) has been created to favor the exchange of knowledge between XI users. Anyone can subscribe to this list and an e-mail has been sent to all the participants of the meeting to make them aware that this list exists and can be used: https://listas.csic.es/cnb/wws/info/3dtomox
Further information about the workshop including the program can be found here

 

One day workshop on Soft X-ray microscopy sample preparation and pre-characterization
Oxford, 7th December 2012

The one-day workshop at the Oxford Particle Imaging Centre of the Wellcome Trust Centre for Human Genetics followed a week of exciting meetings near Oxford, all concerning Soft X-ray cryo-microscopy. Since the five participants of the one day workshop had attended the preceeding 2 day INSTRUCT funded meeting at the Diamond Light Source in Harwell discussing aspects of Soft X-ray cryo-imaging of biological cells, the theory during the workshop could be kept to a minimum to allow as much hands-on experience as possible. The workshop included presentation of sample preparation on special HZB-2 grids for the HZB TXM at the beamline U41-XM of the BESSY II electron storage ring in Berlin, recently established technology for pre-characterization of frozen-hydrated samples on BESSY grids and hands-on practice at three stations. The participants were enthusiastic about the workshop, but all agreed that the schedule was ambitious for such a compact workshop and future workshops should be longer. The workshop was sponsored by BioStruct-X and Carl Zeiss Ltd.

 
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BioStruct-X is part of the Capacities Specific Programme Research Infrastructures, Project Number 283570
BioStruct-X contact: biostructx(at)embl-hamburg.de, I. Custic, ph: +49 - 40 89902 124, fax: +49 - 40 89902 149

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