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Updated: Nov 9, 2025

$Microcrystal Electron Diffraction of Small Molecules$

Microcrystal Electron Diffraction of Small Molecules

Published on: March 15, 2021

Depth-of-field extension in optical imaging for rapid crystal screening.

Chen Li¹, Changqin Ding¹, Minghe Li¹

¹Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, IN 47907, USA.

Acta Crystallographica. Section D, Structural Biology

|April 7, 2021

Summary

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Researchers enhanced the depth of field (DoF) 2.8-fold using a micro-retarder array (µRA) for faster nonlinear optical microscopy. This method improves protein crystal screening by detecting more crystals with fewer images.

Area of Science:

Optics and Photonics
Crystallography
Biophysics

Background:

Nonlinear optical microscopy is crucial for imaging protein crystals.
Current methods face limitations in depth of field (DoF), impacting screening efficiency.
Extending DoF can significantly improve throughput for crystal analysis.

Purpose of the Study:

To enhance the depth of field (DoF) of nonlinear optical microscopy.
To develop a method for rapid protein crystal screening.
To assess the effectiveness of a custom micro-retarder array (µRA) for DoF extension.

Main Methods:

Retrofitting a custom-designed micro-retarder array (µRA) into the optical path.
Utilizing second-harmonic generation (SHG) imaging for protein crystal analysis.

Keywords:

crystal screening depth-of-field extension high throughput

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Related Experiment Videos

Last Updated: Nov 9, 2025

$Microcrystal Electron Diffraction of Small Molecules$

Microcrystal Electron Diffraction of Small Molecules

Published on: March 15, 2021

$On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature$

On-Chip Crystallization and Large-Scale Serial Diffraction at Room Temperature

Published on: March 11, 2022

$Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples$

Synchrotron X-ray Microdiffraction and Fluorescence Imaging of Mineral and Rock Samples

Published on: June 19, 2018

Comparing screening efficiency with and without the enhanced DoF.

Main Results:

Achieved a 2.8-fold extension in the depth of field (DoF).
Increased the number of detected protein crystals during screening.
Reduced the number of `z-slices' required for comprehensive screening.
Experimental results showed excellent agreement with theoretical predictions for wavelength-dependence.

Conclusions:

The micro-retarder array (µRA) offers a simple and effective strategy for DoF enhancement.
This approach significantly increases the throughput of nonlinear optical imaging for protein crystal screening.
The method is broadly applicable to existing nonlinear optical imaging techniques.