1. Overall: The National Resource for Automated Molecular Microscopy Abstract The overall mission of the National Resource for Automated Molecular Microscopy (NRAMM) is to develop, test and apply technology aimed at automating and streamlining cryo-electron microscopy (cryoEM) for structural biology. Our goal from the outset was to provide a pipeline for molecular microscopy that puts it on a par with other structural techniques like X-ray crystallography, so that, once suitable samples are in hand, getting to a high resolution 3D map is a straightforward and rapid undertaking. This one time dream is now obtainable for well-behaved samples as a result of many innovations and improvements in hardware and software, including most recently the advent of a new generation of cameras that directly detect electrons and record high frame rate movies allowing for correction of sample movement during image exposure. There is still work to be done however in ensuring that this method can be applied to the most challenging and biologically interesting samples and serve a very large influx of new practitioners of this method. Our Technology Research and Development (TRD) Projects for the next five years are designed to meet the challenges. Our goals are to develop novel approaches and promote their widespread use, as well as to integrate complex technologies into an efficient and effective method. To achieve these goals, we will focus on three Technological Research and Development Projects that encompass both completely new approaches as well as dynamic evolution of our current technologies. In TRD#1, we will seek to address the critical need to improve and automate vitrified sample preparation; develop methods for time resolved vitrification; and greatly accelerate the throughput of negatively stained sample screening. In TRD#2, we will continue to optimize the performance of both high and mid-range instruments, particularly with regard to integrating and assessing the value of several major accessories; we will develop a high-throughput data acquisition pipeline to support negative stain screening; and we will continue to develop enabling tools for data assessment, processing, analysis and reconstruction. Finally, in TRD#3 we will embark on the development of a new automated and streamlined pipeline for support of in-situ molecular microscopy, that is visualizing molecular structures inside cells. The technological themes at the heart of this proposal support our mission of providing an automated and streamlined pipeline for molecular microscopy that provides data of the highest possible quality and promotes the accessibility of the method to the wider scientific community. This mission is driven by close interactions and feedback from Driving Biological projects (DBPs), and further tested and validated by Collaborative and Service Projects (CSPs). We will also continue to work to maintain excellence in the areas of training and dissemination to promote the broadest possible use of these technologies.

Public Health Relevance

Electron microscopy (EM) has become established as an essential tool for studying macromolecular machines that are central to cellular function and thus novel developments in this area have a basic and fundamental relevance for both the healthy and diseased states. This project will develop novel technologies and increase the pace and reach of EM structural studies driven by fundamental research efforts in basic science, drug and vaccine development.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Flicker, Paula F
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
New York Structural Biology Center
New York
United States
Zip Code
Singh, Appu K; McGoldrick, Luke L; Sobolevsky, Alexander I (2018) Structure and gating mechanism of the transient receptor potential channel TRPV3. Nat Struct Mol Biol 25:805-813
Kang, Jin Young; Mooney, Rachel Anne; Nedialkov, Yuri et al. (2018) Structural Basis for Transcript Elongation Control by NusG Family Universal Regulators. Cell 173:1650-1662.e14
Benoit, Matthieu P M H; Asenjo, Ana B; Sosa, Hernando (2018) Cryo-EM reveals the structural basis of microtubule depolymerization by kinesin-13s. Nat Commun 9:1662
Sun, Yadong; Zhang, Yixiao; Hamilton, Keith et al. (2018) Molecular basis for the recognition of the human AAUAAA polyadenylation signal. Proc Natl Acad Sci U S A 115:E1419-E1428
Wei, Hui; Dandey, Venkata P; Zhang, Zhening et al. (2018) Optimizing ""self-wicking"" nanowire grids. J Struct Biol 202:170-174
Piasecka, Anna; Czapinska, Honorata; Vielberg, Marie-Theres et al. (2018) The Y. bercovieri Anbu crystal structure sheds light on the evolution of highly (pseudo)symmetric multimers. J Mol Biol 430:611-627
Noble, Alex J; Dandey, Venkata P; Wei, Hui et al. (2018) Routine single particle CryoEM sample and grid characterization by tomography. Elife 7:
Singh, Appu K; McGoldrick, Luke L; Twomey, Edward C et al. (2018) Mechanism of calmodulin inactivation of the calcium-selective TRP channel TRPV6. Sci Adv 4:eaau6088
Dingens, Adam S; Acharya, Priyamvada; Haddox, Hugh K et al. (2018) Complete functional mapping of infection- and vaccine-elicited antibodies against the fusion peptide of HIV. PLoS Pathog 14:e1007159
Lin, Wei; Das, Kalyan; Degen, David et al. (2018) Structural Basis of Transcription Inhibition by Fidaxomicin (Lipiarmycin A3). Mol Cell 70:60-71.e15

Showing the most recent 10 out of 159 publications