Detachable Nozzle Piezoelectric Dispense System for Cryo-TEM Sample Preparation

用于冷冻 TEM 样品制备的可拆卸喷嘴压电分配系统

• 批准号: 8590991
• 负责人: Peter Kahn
• 金额: $ 14.98万
• 依托单位: ENGINEERING ARTS, LLC
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2015-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8590991
• 关键词: Arts; Binding; Biological; Biotechnology; Buffers; Caliber; Communities; Consumption; Cucumber; Custom; Development; Diagnostic; Disease; Electron Microscopy; Electronics; Engineering; Ethane; Event; Flow Cytometry; Foundations; Freezing; Grant; Head; Ice; Image; Lead; Legal patent; Letters; Licensing; Life; Lipids; Liquid substance; Methods; Microscopy; Microtubules; Molecular; Molecular Models; Nanotechnology; Nanotubes; Necrosis; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Preparation; Process; Productivity; Pump; Research; Research Infrastructure; Research Institute; Resolution; Resources; Sales; Sampling; Scientist; Services; Specimen; Structure; Syringes; System; Testing; Thick; Time; Transmission Electron Microscopy; Validation; Viral Proteins; Virus; Work; abstracting; aqueous; base; college; commercial application; commercialization; design; improved; innovation; insight; instrument; instrumentation; interest; macromolecule; molecular modeling; nanoimaging; nanomaterials; nanoparticle; open source; prototype; public health relevance; scale up; success; technological innovation; technology development; time use; tool

DESCRIPTION (provided by applicant): Project Summary / Abstract Engineering Arts (EA) proposes the development of a detachable nozzle piezoelectric dispense system for use in Cryo-TEM (Cold - Transmission Electron Microscopy) sample preparation instrumentation. Cryo-TEM is an important tool for studying the structure of biological macromolecules in their native aqueous state. The challenging and laborious process of preparing 'thin-ice' frozen aqueous samples for Cryo-TEM has remained essentially unchanged for decades. Typically a single aqueous biological sample (~3¿l) is manually applied onto a thin porous Electron Microscopy (EM) grid which is then blotted to remove excess sample and then plunged into liquid-ethane for flash freezing. This process hopefully leaves enough regions on the grid with the proper vitrified 'thin-ice', typically 50 to 200nm thickness, required for EM imaging. The detachable piezoelectric nozzle dispense system proposed here is a key innovation that will lead to productivity, quality and capability breakthroughs for Cryo-TEM workflow. The following key benifits for Cryo-TEM process will be realized: 1. Apply many different samples (or sample conditions) to each Cryo-TEM grid. 2. Increase process control and yields of acceptable thin-ice over current standard blotting methods. 3. Lower starting sample volume requirements and consumption per grid. 4. Allow time-resolved study of mixing / binding events of multiple samples. EA will design, fabricate and optimize a multichannel detachable nozzle piezoelectric dispense system that is capable of dispensing droplets (less than 50 picoliters) of several different biological samples (or sample conditions) onto each TEM grid. A three channel detachable nozzle piezoelectric dispense system retrofitted onto a custom Cryo-TEM sample preparation instrument (Spotiton instrument platform) at NRAMM (National Resource for Automated Molecular Microscopy, Scripps, La Jolla CA.) for testing, optimization and validation.

描述（由申请人提供）：项目摘要/摘要工程艺术（EA）提出开发用于低温透射电子显微镜（Cryo-TEM）样品制备仪器的可拆卸喷嘴压电分配系统。低温透射电镜是研究生物大分子水溶液结构的重要工具。为Cryo-TEM制备“薄冰”冷冻水溶液样品的挑战性和费力的过程几十年来基本上保持不变。通常，将单个含水生物样品（约3升）手动施加到薄的多孔电子显微镜（EM）网格上，然后将其吸干以去除多余的样品，然后将其浸入液体乙烷中进行快速冷冻。这个过程有希望在网格上留下足够的区域，具有适当的玻璃化“薄冰”，通常为50至200 nm的厚度，用于EM成像。本文提出的可拆卸压电喷嘴点胶系统是一项关键创新，将为Cryo-TEM工作流程带来生产率、质量和能力的突破。Cryo-TEM工艺将实现以下主要优点：1。将许多不同的样品（或样品条件）应用于每个Cryo-TEM网格。2.与现有标准印迹方法相比，提高过程控制和可接受的薄冰产量。3.更低的起始样品体积要求和每个网格的消耗。4.允许对多个样品的混合/结合事件进行时间分辨研究。EA将设计，制造和优化多通道可拆卸喷嘴压电分配系统，该系统能够将几种不同生物样品（或样品条件）的液滴（小于50皮升）分配到每个TEM网格上。在NRAMM（National Resource for Automated Molecular Microscopy，Scripps，拉霍亚CA.）用于测试、优化和验证。