Next generation sample preparation for cryo-EM based on Spotiton
blot-free, high-speed plunging
reduce air-water interface effects and enable difficult samples
easy to use even for novice users
guided workflows enables simple set up, use and cleaning
record keeping for future repeatability
capture all relevant grid parameters and images
screen fewer grids at the microscope
select good grids based on visual images during freezing
virtually no manual handling
eliminate damage and loss via precise automated grid handling
safe storage and control of cryogens
automated drawer offers level sensing and temperature control
The sample preparation bottleneck...
Recent advances in cryo-EM have led to an explosion of interest in the technique for increasingly challenging structural biology projects. Producing quality frozen grids with vitreous ice of an appropriate thickness and evenly distributed particles remains one of the major bottlenecks in the structure determination process. Currently, highly skilled manual handling needs to be learnt to avoid physical grid damage. Poor traceability of storage locations can also lead to grid losses. Feedback on ice quality requires costly screening on an electron microscope, time better used for more productive data collection. chameleon’s aim is to enable automated, rapid progress to the optimum ice thickness for a range of samples, even those handled by researchers relatively new to cryo-EM.
The chameleon story...
chameleon is a commercial development of the successful Spotiton system, an idea conceived and brought to life by the groups of Bridget Carragher and Clint Potter at New York Structural Biology Centre (NYSBC). Realising the limitations of existing grid preparation practices, they set out to develop a robust and repeatable technique for specimen vitrification based on small volume dispensing using inkjet technology. The 'Spotition' (first published in 2012) as it became known, then underwent a series of iterative improvements in-house and became instrumental in an increasing number of high quality structures and cutting edge research papers on air water interface effects published by the team. chameleon distils out the advatages of Spotition and builds in our proven knowledge of process automation in the field of structural biology.
Self-wicking grids for cryo-EM
Key development references:
Spotiton: A prototype for an integrated inkjet dispense and vitrification system for cryo-TEM
Tilak Jain, Patrick Sheehan, John Crum, Bridget Carragher, Clinton S.Potter
A new method for vitrifying samples for cryoEM
Ivan Razinkov,Venkata P. Dandey, Hui Wei Zhening Zhang, David Melnekoff, William J.Rice, Christoph Wigge, Clinton S.Potter, Bridget Carragher
Spotiton: New features and applications
Venkata P. Dandey, Hui Weia Zhening Zhang, Yong Zi Tan, Priyamvada Acharya, Edward T. Eng, William J. Rice, Peter A.Kahn, Clinton S.Potter, Bridget Carragher
Optimizing “self-wicking” nanowire grids
Hui Wei, Venkata P. Dandey, Zhening Zhang, Ashleigh Raczkowski, Willam J. Rice, Bridget Carragher, Clinton S.Potter
Reducing effects of particle adsorption to the air-water interface in cryo-EM
Alex J. Noble, Hui Wei, Venkata P. Dandey, Zhening Zhang, Yong Zi Tan, Clinton S.Potter & Bridget Carragher,
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