TY - CHAP
T1 - Automation in Single-Particle Electron Microscopy. Connecting the Pieces
AU - Lyumkis, Dmitry
AU - Moeller, Arne
AU - Cheng, Anchi
AU - Herold, Amber
AU - Hou, Eric
AU - Irving, Christopher
AU - Jacovetty, Erica L.
AU - Lau, Pick Wei
AU - Mulder, Anke M.
AU - Pulokas, James
AU - Quispe, Joel D.
AU - Voss, Neil R.
AU - Potter, Clinton S.
AU - Carragher, Bridget
N1 - Funding Information:
We acknowledge primary support from the National Institutes of Health (NIH) through the National Center for Research Resources (NCRR) P41 program (Grants RR17573 and RR023093). We are also grateful for the support provided to Arne Moeller by the Joint Center for Innovation in Membrane Protein Production for Structure Determination (Grant RFA-RM-08-019) and to Pick-Wei Lau by the American Heart Association. We ask for forgiveness for any omissions, errors, or oversights that are almost inevitable in a review of this wide scope.
PY - 2010
Y1 - 2010
N2 - Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the size of collected data sets, comprehensive algorithms for image processing, sophisticated tools for quality assessment, and an influx of interested scientists from outside the field who might lack the skills of experienced microscopists. This situation places automated techniques in high demand. In this chapter, we provide a generic definition of and discuss some of the most important advances in automated approaches to specimen preparation, grid handling, robotic screening, microscope calibrations, data acquisition, image processing, and computational infrastructure. Each section describes the general problem and then provides examples of how that problem has been addressed through automation, highlighting available processing packages, and sometimes describing the particular approach at the National Resource for Automated Molecular Microscopy (NRAMM). We contrast the more familiar manual procedures with automated approaches, emphasizing breakthroughs as well as current limitations. Finally, we speculate on future directions and improvements in automated technologies. Our overall goal is to present automation as more than simply a tool to save time. Rather, we aim to illustrate that automation is a comprehensive and versatile strategy that can deliver biological information on an unprecedented scale beyond the scope available with classical manual approaches.
AB - Throughout the history of single-particle electron microscopy (EM), automated technologies have seen varying degrees of emphasis and development, usually depending upon the contemporary demands of the field. We are currently faced with increasingly sophisticated devices for specimen preparation, vast increases in the size of collected data sets, comprehensive algorithms for image processing, sophisticated tools for quality assessment, and an influx of interested scientists from outside the field who might lack the skills of experienced microscopists. This situation places automated techniques in high demand. In this chapter, we provide a generic definition of and discuss some of the most important advances in automated approaches to specimen preparation, grid handling, robotic screening, microscope calibrations, data acquisition, image processing, and computational infrastructure. Each section describes the general problem and then provides examples of how that problem has been addressed through automation, highlighting available processing packages, and sometimes describing the particular approach at the National Resource for Automated Molecular Microscopy (NRAMM). We contrast the more familiar manual procedures with automated approaches, emphasizing breakthroughs as well as current limitations. Finally, we speculate on future directions and improvements in automated technologies. Our overall goal is to present automation as more than simply a tool to save time. Rather, we aim to illustrate that automation is a comprehensive and versatile strategy that can deliver biological information on an unprecedented scale beyond the scope available with classical manual approaches.
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U2 - 10.1016/S0076-6879(10)83015-0
DO - 10.1016/S0076-6879(10)83015-0
M3 - Chapter
C2 - 20888480
AN - SCOPUS:77957264753
T3 - Methods in Enzymology
SP - 291
EP - 338
BT - Methods in Enzymology
PB - Academic Press Inc.
ER -