A new method for vitrifying samples for cryoEM.
Almost every aspect of cryo electron microscopy (cryoEM) has been automated over the last few decades. One of the challenges that remains to be addressed is the robust and reliable preparation of vitrified specimens of suitable ice thickness. We present results from a new device for preparing vitrified samples. The successful use of the device is coupled to a new "self-blotting" grid that we have developed to provide a method for spreading a sample to a thin film without the use of externally applied filter paper. This new approach has the advantage of using small amounts of protein material, resulting in large areas of ice of a well defined thickness containing evenly distributed single particles. We believe that these methods will in the future result in a system for vitrifying grids that is completely automated.
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Ivan
I
Razinkov
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Venkat
V
Dandey
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Hui
H
Wei
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Zhening
Z
Zhang
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
David
D
Melnekoff
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
William J
WJ
Rice
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Christoph
C
Wigge
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Clinton S
CS
Potter
Simons Electron Microscopy Center, The National Resource for Automated Molecular Microscopy, New York Structural Biology Center.
Bridget
B
Carragher
D020285:Cryoelectron Microscopy / Q000295:instrumentation* / Q000379:methods; D007053:Ice / Q000032:analysis; D007091:Image Processing, Computer-Assisted; D013048:Specimen Handling / Q000295:instrumentation* / Q000379:methods; D058989:Vitrification
D007053:Ice
Automation; CryoTEM; Nanowires; Self-blotting grids