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  • Open Access

Noninvasive, Label-Free Imaging of Freely Moving C. elegans Nematodes

Rongbiao Tang1,*, Feixiang Wang2, Ke Li2, and Yanan Fu2

  • *Contact author: tangme8688258@126.com
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PRX Life 3, 043019 – Published 17 December, 2025

DOI: https://doi.org/10.1103/j9bg-ntxw

Abstract

Noninvasive, label-free imaging of freely moving Caenorhabditis elegans nematodes offers a powerful but technically demanding approach for behavioral and morphological studies. Key challenges arise from their microscopic size, rapid and intricate locomotion patterns, uniform soft-tissue composition with low intrinsic contrast, and the stringent requirements for imaging systems capable of simultaneously achieving high spatiotemporal resolution and exceptional soft-tissue sensitivity. To overcome these challenges, we employed ultrafast phase-contrast x-ray imaging (PCXI) and phase-contrast computed tomography (PCCT). PCXI was conducted at a detector acquisition speed of 500 fps, while PCCT leveraged a synchronized system comprising a fast-acquisition detector (4000 fps), a rapid-rotation stage (1800∘/s), and a precision electromechanical shutter (3-ms actuation). This configuration enabled complete three-dimensional (3D) tomographic acquisition (400 projections over 180∘) in just 100 ms. Our integrated imaging platform delivers dual capabilities: PCXI enables real-time monitoring of multiple nematodes' natural behaviors, including dynamic irradiation avoidance responses, while PCCT offers comprehensive whole-organism morphometric analysis—quantifying body length, thickness, and volume—all under physiologically natural conditions without the need for anesthesia or contrast agent staining. By synergistically integrating ultrafast PCXI for dynamic two-dimensional functional analysis with PCCT for comprehensive 3D morphological characterization, this multimodal platform establishes an imaging paradigm that significantly advances the study of fast-moving microscopic organisms.

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PRX Life 3, 043019– Published 17 December, 2025
  • Received 9 July 2025
  • Accepted 3 November 2025
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Published by the American Physical Society under the terms of the Creative Commons Attribution 4.0 International license. Further distribution of this work must maintain attribution to the author(s) and the published article's title, journal citation, and DOI.

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