Visualizing cellular ultrastructure using light microscopy in hematology

在血液学中使用光学显微镜观察细胞超微结构

基本信息

  • 批准号:
    10473885
  • 负责人:
  • 金额:
    $ 20.5万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2021
  • 资助国家:
    美国
  • 起止时间:
    2021-08-25 至 2024-07-31
  • 项目状态:
    已结题

项目摘要

Project Summary/Abstract Imaging is an important diagnostic and research tool in hematology. This R21 proposal, written in response to PAR-20-140 (Catalytic Tool and Technology Development in Kidney, Urologic, and Hematologic Diseases), is designed to optimize, validate, and deploy a groundbreaking light microscopy platform for visualization of proteins in their ultrastructural context in hematology. Visualizing specific proteins of interest in their ultrastructural (sub-organelle, ~sub-50 nm) context currently requires correlative light and electron microscopy (CLEM) which is extremely complicated, time-consuming and expensive and thereby inaccessible to most researchers in hematology. Pan Expansion Microscopy (PanExM) is a novel imaging approach that allows for imaging of proteins (and other biomolecules and organelles) in their ultrastructural context on a whole-cell basis with nanoscale (~10-30 nm) resolution using conventional (diffraction-limited) light microscopes. With PanExM, fixed cells are expanded up to 20-fold in the linear dimension (~8000 fold in volume) while maintaining subcellular organization, cellular protein content and antigenicity. Staining the preserved ultrastructure as well as the protein of interest in these samples in different colors and imaging them in three dimensions using standard light microscopy allows for visualization and analysis of what would otherwise be below the diffraction limit of light. Aim 1 is focused on developing and optimizing the methodology to expand individual blood and bone marrow cells adhered or immobilized on slides. Aim 2 is focused on optimizing expansion of bone marrow sections to assess hematopoietic cells in their microenvironment. Our highly synergistic research team is dedicated to training and dissemination of the latest technologies in hematology research. Application of this powerful new technology to blood and bone marrow cells will be transformative by enabling hematology investigators to quantify the location, distribution and interactions of molecules of interest in their sub-cellular context. By extending these high-resolution imaging capabilities from a handful to hundreds of research groups and increasing the throughput from weeks per data set to hours, PanExM will democratize the hematology research community’s access to 3D structural information and will drive discoveries across biomedical research ranging from fundamental cell biological findings to histological applications in clinical pathology.
项目总结/摘要 影像学是血液学中重要的诊断和研究工具。这份R21提案是为了回应 PAR-20-140(肾脏、泌尿系统和血液系统疾病的催化工具和技术开发), 旨在优化,验证和部署一个突破性的光学显微镜平台,用于可视化 蛋白质在血液学中的超微结构背景。可视化特定的感兴趣的蛋白质, 超微结构(亚细胞器,~亚50 nm)背景目前需要相关的光学和电子显微镜 (CLEM),其极其复杂、耗时且昂贵,因此大多数人无法使用。 血液学研究者泛扩展显微镜(PanExM)是一种新的成像方法,允许 在全细胞的基础上,对蛋白质(以及其他生物分子和细胞器)进行超微结构成像 使用常规(衍射极限)光学显微镜以纳米级(~10-30 nm)分辨率测量。通过PanExM, 固定的细胞在线性尺寸上扩增高达20倍(体积约8000倍),同时保持亚细胞 组织、细胞蛋白含量和抗原性。染色保存的超微结构以及蛋白质 并使用标准光对这些样品进行三维成像 显微镜允许观察和分析否则将低于光的衍射极限的东西。 目的1是发展和优化扩增个体血液和骨髓的方法学 细胞粘附或固定在载玻片上。目标2的重点是优化骨髓切片的扩增, 评估微环境中的造血细胞。我们高度协同的研究团队致力于 培训和传播血液学研究的最新技术。应用这一强大的新 血液和骨髓细胞的技术将通过使血液学研究者能够 量化目标分子在其亚细胞环境中的位置、分布和相互作用。通过 将这些高分辨率成像能力从少数扩展到数百个研究小组, 通过将每个数据集的处理量从数周增加到数小时,PanExM将使血液学研究民主化 社区对3D结构信息的访问,并将推动生物医学研究的发现, 从基本的细胞生物学发现到临床病理学中的组织学应用。

项目成果

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Diane S Krause其他文献

Diane S Krause的其他文献

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{{ truncateString('Diane S Krause', 18)}}的其他基金

Visualizing cellular ultrastructure using light microscopy in hematology
在血液学中使用光学显微镜观察细胞超微结构
  • 批准号:
    10316778
  • 财政年份:
    2021
  • 资助金额:
    $ 20.5万
  • 项目类别:
"Exploration of Human Parathyroid Cellular Organization and Function"
《人类甲状旁腺细胞组织与功能的探索》
  • 批准号:
    10044664
  • 财政年份:
    2020
  • 资助金额:
    $ 20.5万
  • 项目类别:
Megakaryocyte erythroid progenitor fate specification
巨核细胞红系祖细胞命运规范
  • 批准号:
    9764359
  • 财政年份:
    2017
  • 资助金额:
    $ 20.5万
  • 项目类别:
Megakaryocyte erythroid progenitor fate specification
巨核细胞红系祖细胞命运规范
  • 批准号:
    9363263
  • 财政年份:
    2017
  • 资助金额:
    $ 20.5万
  • 项目类别:
Megakaryocyte erythroid progenitor fate specification
巨核细胞红系祖细胞命运规范
  • 批准号:
    10001510
  • 财政年份:
    2017
  • 资助金额:
    $ 20.5万
  • 项目类别:
Administrative Core
行政核心
  • 批准号:
    10060456
  • 财政年份:
    2015
  • 资助金额:
    $ 20.5万
  • 项目类别:
Administrative Core
行政核心
  • 批准号:
    10677844
  • 财政年份:
    2015
  • 资助金额:
    $ 20.5万
  • 项目类别:
Administrative Core
行政核心
  • 批准号:
    10249340
  • 财政年份:
    2015
  • 资助金额:
    $ 20.5万
  • 项目类别:
Cell Preparation and Analysis Core
细胞制备和分析核心
  • 批准号:
    10677841
  • 财政年份:
    2015
  • 资助金额:
    $ 20.5万
  • 项目类别:
Administrative Core
行政核心
  • 批准号:
    10624200
  • 财政年份:
    2015
  • 资助金额:
    $ 20.5万
  • 项目类别:

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