# Single-cell structural biology with intracellular electron crystallography

## TL;DR

IncelluloED is a new method that allows high-resolution protein structure determination from a single cell using electron diffraction, reducing the need for large sample volumes.

## Contribution

IncelluloED introduces a novel method for high-resolution structural biology using a single intracellular crystal and 3D electron diffraction.

## Key findings

- IncelluloED achieved a 1.9 Å resolution structure from a single cell's microcrystal.
- The method requires a much smaller sample volume compared to serial X-ray crystallography.
- IncelluloED uses widely available cryo-EM tools, making high-resolution structural studies accessible to home laboratories.

## Abstract

Intracellular crystallization is an emerging approach in structural biology that bypasses the need for protein purification. In 2024, the InCellCryst pipeline was introduced for structural studies of intracellular crystals by serial X-ray crystallography. Serial crystallography requires the exposure of tens of thousands of cells containing intracellular crystals, precluding high-resolution studies on proteins that crystallize only in a few cells. Here we introduce IncelluloED, a method that combines intracellular crystallization with in situ 3D electron diffraction in cells and achieves high-resolution structures from just one crystal inside one cell. Experiments on a microcrystal of the HEX-1 protein from Magnaporthe grisea, grown inside an insect cell, give a structure at 1.9 Å resolution from a volume of ~1.6 µm3 as compared to 1.8 Å resolution achieved by serial X-ray crystallography from a combined volume exceeding eleven million µm3. IncelluloED uses widely available cryo-EM tools and brings high-resolution structural biology into home laboratories while also advancing a vision for a “single-cell structural laboratory”.

IncelluloED combines intracellular crystallization with 3D electron diffraction, enabling high-resolution structural studies of a single protein crystal inside a cell using cryo-EM tools, and paving the way toward a single-cell structural laboratory.