Pre-session information

Shipping information

Our address for sending dry shippers in advance:

MVLS Urquhart Stores,
University of Glasgow,
464 Bearsden Road,
Glasgow,
G61 1QH

If you are coming to SCMI in person for your session, you are also welcome to bring your specimens on the day.

Travelling to SCMI and Parking

SCMI is located in the Centre for Virus Research (CVR) Sir Michael Stoker Building in the University of Glasgow Garscube Campus

 

Travelling to SCMI by Public Transport

 

Travelling to Garscube Campus by car

The main vehicular access to the Garscube Campus is via Switchback Road. If using a navigation app/Sat Nav please use G61 1BD as the postcode; this will take you directly to the Campus Gatehouse where staff will be able to direct you to the Sir Michael Stoker Building.  If you plan to travel by car please contact our facility manager James Streetley in advance and he will arrange a parking space for you.

 

Pixel size and camera set-up

It is worth thinking about how you want the microscope and camera to be configured in advance of your session. You can always discuss this with the staff setting up your session, but having one or two ideas will help speed it up.

Linear or counting

The DE-64 can run in two modes; linear or counting. Counting has a higher DQE but we have to hardware bin the camera by a factor of two. This means that for a given pixel size, a linear image is 4 times larger. So the trade-off is field of view vs. quality. Counting is also slower than linear.

Pixel size (or magification)

Small pixel sizes (high magnifications) are needed to achieve high resolution reconstructions. However, that doesn't mean the smallest pixel size is appropriate for every project. If your complex is sparse or large then the small field of view at high magnifications may not give you very many particles (also a reason to consider linear mode). Equally, if you expect the resolution of your final reconstruction to be limited by some other reason, then it does not make sense to use a very small pixel size. It would be better to collect more particles at a larger pixel size.

Common pixel sizes in linear
Magnification

Pixel size / Å

40,000x

1.499

50,000x

1.177

60,000x

0.997
Common pixel sizes in counting
MagnificationPixel size / Å
100,000x

1.209

120,000x

1.015

150,000x

0.829

200,000x

0.598

 

Training Resources

Video Tutorials and Seminars

Getting Started in Cryo-EM  A series of tutorials from Professor Grant Jensen at Caltech

LMB Cryo-EM course MRC Laboratory for Molecular Biology 2017 Cryo-EM course.

 

 

Useful Publications

An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology.

Rebecca F. Thompson, Matt Walker, C. Alistair Siebert, Stephen P. Muench, Neil A. Ranson

http://dx.doi.org/10.1016/j.ymeth.2016.02.017

 

A primer to Single-Particle Cryo-Electron Microscopy.

Yifang Chen, Nikolous Grigorieff, Pawel A. Penczek, Thomas Walz

http://dx.doi.org/10.1016/j.cell.2015.03.050

 

 

Session Timings

CRYO ARM 300 data collection sessions run for 48 or 72 hours. The user is only requested to be on site for the first working day of the session. An approximate session is outlined below, although this is dependent upon the requested microscope configuration and the specimen.

 

UK time24h time 
9.30am 09:30 Arrive at SCMI
9.30am - 10am 09:30 - 10:00 Load samples
10am - 12pm 10:00 - 12:00 Set up imaging conditions, screen grids, take test images
12pm - 1pm 12:00 - 13:00 Gain Reference (if required)
1pm - 2pm 13:00 - 14:00 Collect grid square montages
2pm - 3pm 14:00 - 15:00 Select targets for acquisition
3pm 15:00 Run session, set up data transfer. Adjust settings if necessary