SBIR-STTR Award

Development and Commercialization of a Sample Preparation System for Time Resolved Cryo-Electron Microscopy Company Name: Hummingbird Precision Machine Co., dba Hummingbird Scientific Principal Investigator: Norman Salmon Summary: Cryo-electron microscopy (cryo-EM) is a powerful technique for imaging biological molecules in which samples are preserved in a frozen, hydrated state before imaging using an electron beam. The technique provides an alternative to X-ray crystallography for structural characterization of biological samples that are not crystallizable or do not crystallize in their native state. While cryo- EM sample preparation technique has become more efficient in recent years with the development of automatic sample vitrification systems to perform grid preparation, the field of cryo-EM is hindered by the inability to perform time-resolved experiments on a sufficiently fast time scale to observe the progression of many biological reactions due to the inherent lag of traditional blotting methods. The Frank group at Columbia University has developed a lab- prototype of a system to enable this missing capability and enables millisecond time control over the reaction time of sampling biological reactions in the grid preparation process. The purpose of this Fast-Track STTR proposal is to develop the time-resolved system built by the Frank group into a commercial production unit and launch it as a product. Initial Phase I milestones in the proposed work will focus on building a prototype with added control of reaction parameters to determine necessary control variables in the system and to optimize sample grid preparation and time resolution. Further work in Phase II of this Fast-Track proposal will create a streamlined, user-friendly production unit which will maintain the necessary controls as determined in initial milestones, while optimizing manufacturing time and reducing overall production costs. The resulting system will add another dimension to the field of cryo-EM, providing users with the capability to capture transient intermediates in reaction progression studies and further clarifying proposed reaction mechanisms with time resolution.

Public Health Relevance Statement:
Project Narrative Project Title: Development and Commercialization of a Sample Preparation System for Time Resolved Cryo-Electron Microscopy Company Name: Hummingbird Precision Machine Co., dba Hummingbird Scientific Principal Investigator: Norman Salmon Narrative: Particle structure and relative population determination are key components in biological research, informing predictions of reaction mechanisms in biological processes, whereas reaction mechanisms, give a greater understanding of different biological pathways that direct organismal functions. The proposal develops a time-resolved cryo-electron microscopy method and hardware that brings new possibilities to structural biological research by allowing researchers to study the evolution of population distributions as reactions progresses. It also enables studies into the structure of any intermediate states that are too fast-lived to be captured by traditional pipetting-blotting cryo-electron microcopy sample preparation techniques, all in the native, hydrated state of the molecule.

Project Terms:
Biological; Biological Process; biological research; biological systems; commercialization; Cone; Consumption; cost; Cost efficiency; Cryoelectron Microscopy; Data; design; Development; Dimensions; Documentation; Electron Beam; Electrons; Environmental Monitoring; Ethane; Evaluation; Evolution; experimental study; Freezing; Gases; Government; Humidity; Hydration status; Image; Imaging Techniques; improved; Kinetics; Liquid substance; Measures; Methods; Microfluidics; millisecond; Molecular Structure; Names; operation; particle; Pathway interactions; Persons; Phase; Population; Population Distributions; Positioning Attribute; Preparation; preservation; pressure; Principal Investigator; Process; Production; prototype; Reaction; Reaction Time; Recipe; Reproducibility; Research; Research Personnel; Resolution; Running; Salmon; Sampling; Small Business Technology Transfer Research; software development; Speed; Standardization; Structure; System; Techniques; Temperature; temporal measurement; Testing; Time; touchscreen; Universities; user-friendly; Work; X-Ray Crystallography

Cryo-electron microscopy (cryo-EM) is a powerful technique for imaging biological molecules inwhich samples are preserved in a frozen, hydrated state before imaging using an electron beam.The technique provides an alternative to X-ray crystallography for structural characterization ofbiological samples that are not crystallizable or do not crystallize in their native state. While cryo-EM sample preparation technique has become more efficient in recent years with thedevelopment of automatic sample vitrification systems to perform grid preparation, the field ofcryo-EM is hindered by the inability to perform time-resolved experiments on a sufficiently fasttime scale to observe the progression of many biological reactions due to the inherent lag oftraditional blotting methods. The Frank group at Columbia University has developed a lab-prototype of a system to enable this missing capability and enables millisecond time control overthe reaction time of sampling biological reactions in the grid preparation process.The purpose of this Fast-Track STTR proposal is to develop the time-resolved system built by theFrank group into a commercial production unit and launch it as a product. Initial Phase Imilestones in the proposed work will focus on building a prototype with added control of reactionparameters to determine necessary control variables in the system and to optimize sample gridpreparation and time resolution. Further work in Phase II of this Fast-Track proposal will create astreamlined, user-friendly production unit which will maintain the necessary controls asdetermined in initial milestones, while optimizing manufacturing time and reducing overallproduction costs. The resulting system will add another dimension to the field of cryo-EM,providing users with the capability to capture transient intermediates in reaction progressionstudies and further clarifying proposed reaction mechanisms with time resolution.

Public Health Relevance Statement:
Particle structure and relative population determination are key components in biological research, informing predictions of reaction mechanisms in biological processes, whereas reaction mechanisms, give a greater understanding of different biological pathways that direct organismal functions. The proposal develops a time-resolved cryo-electron microscopy method and hardware that brings new possibilities to structural biological research by allowing researchers to study the evolution of population distributions as reactions progresses. It also enables studies into the structure of any intermediate states that are too fast-lived to be captured by traditional pipetting-blotting cryo-electron microcopy sample preparation techniques, all in the native, hydrated state of the molecule.

Project Terms:
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