# Visualizing cellular ultrastructure using light microscopy in hematology

> **NIH NIH R21** · YALE UNIVERSITY · 2021 · $244,148

## Abstract

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.

## Key facts

- **NIH application ID:** 10316778
- **Project number:** 1R21DK130299-01
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Diane S Krause
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $244,148
- **Award type:** 1
- **Project period:** 2021-08-25 → 2023-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10316778

## Citation

> US National Institutes of Health, RePORTER application 10316778, Visualizing cellular ultrastructure using light microscopy in hematology (1R21DK130299-01). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10316778. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*