# Advancing Multi-Color EM via Direct Detector-enabled 4D-STEM

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA, SAN DIEGO · 2021 · $353,911

## Abstract

PROJECT SUMMARY
This focused technology development project will advance into first practice a transformative new imaging
technology for multicolor electron microscopy (ColorEM), enabling new capabilities for high resolution and 3
dimensional (3D) localization and differentiation of different molecular complexes in cells and tissues. With this
technology, we will substantively improve upon presently practiced, multiple labeling strategies involving
antibody labelling approaches detected using colloidal gold, quantum dots, etc.. We will further provide a more
accessible, higher throughput, and dose efficient alternative to spectroscopic methods, like energy dispersive
x-ray (EDX) and electron energy loss spectroscopy (EELS) / energy filtered transmission electron
microscopy (EFTEM). At the heart of this effort, we will exploit the performance characteristics of a next-
generation, ultra-high speed direct detection device (DDD), now being optimized and extended (by us with an
industry partner) for use with scanning transmission electron microscopy (STEM), a broadly deployed
capability on modern EM columns. The optimized application of this new pixelated STEM detector
technology, in combination with an extended palette of elemental probes and advanced software analysis,
will allow for unprecedented new capabilities for performing atomic number (Z) contrast imaging – a
process we call ColorSTEM. To bring this new integrated methodology into first practice, we will build on
recent feasibility studies and surmount remaining technical hurdles to 1) make multi-labelled EM specimens
with optimized elemental probe combinations; 2) troubleshoot the first use of a potentially transformative new
sensor technology and work out the methodology for employing it for Z contrast imaging; and 3) devise the
associated analytical/computational methodology for processing the very large resultant data sets to enable
unambiguous differentiation of molecular complexes selectively marked with elemental probes.

## Key facts

- **NIH application ID:** 10248396
- **Project number:** 5R01GM138780-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA, SAN DIEGO
- **Principal Investigator:** Mark H Ellisman
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $353,911
- **Award type:** 5
- **Project period:** 2020-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10248396

## Citation

> US National Institutes of Health, RePORTER application 10248396, Advancing Multi-Color EM via Direct Detector-enabled 4D-STEM (5R01GM138780-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10248396. Licensed CC0.

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