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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：
10316778
负责人：
Diane S Krause
金额：
$ 24.41万
依托单位：
YALE UNIVERSITY
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 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10316778
关键词：
3-Dimensional Address Algorithmic Analysis Antibodies Architecture Archives Biological Biomedical Research Blood Blood Cells Blood Platelets Bone Marrow Bone Marrow Cells Cells Clinical Pathology Collaborations Color Communities Complex Complex Analysis Confocal Microscopy Consumption Data Set Development Diagnostics Research Dimensions Dyes Electron Microscopy Environment Fluorescence Fluorescence Microscopy Goals Head Hematological Disease Hematology Hematopoietic Histologic Hour Human Image Imaging technology Immobilization Immunofluorescence Immunologic Individual Investigation Isotropy Kidney Diseases Knowledge Letters Light Light Microscope Location Mammalian Cell Marrow Megakaryocytes Methodology Microscope Microscopy Molecular Organelles Pathology Proteins Protocols documentation Research Research Personnel Resolution Sampling Slide Stains Subcellular structure Swelling Techniques Technology Time Tissues Training Urologic Diseases Vision Visualization cell type cellular imaging design high resolution imaging imaging approach imaging capabilities improved insight interest light microscopy macrophage nanoscale neglect new technology novel peripheral blood preservation protein distribution response technology development three dimensional structure tool tool development

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/文摘