posted on 2025-08-01, 12:44authored bySL Burg, AL Washington, J Villanova, AJC Dennison, D McLoughlin, OO Mykhaylyk, P Vukusic, W Furnass, RAL Jones, AJ Parnell, JPA Fairclough
High resolution X-ray nano-tomography experiments are often limited to a few tens of micrometer size volumes due to detector size. It is possible, through the use of multiple overlapping tomography scans, to produce a large area scan which can encompass a sample in its entirety. Mounting and positioning regions to be scanned is highly challenging and normally requires focused ion beam approaches. In this work we have imaged intact beetle scale cells mounted on the tip of a needle using a micromanipulator stage. Here we show X-ray holotomography data for single ultra-white scales from the beetles Lepidiota stigma (L. stigma) and Cyphochilus which exhibit the most effective scattering of white light in the literature. The final thresholded matrices represent a scan area of 25 × 70 × 362.5 µm and 25 × 67.5 × 235µm while maintaining a pixel resolution of 25 nm. This tomographic approach allowed the internal structure of the scales to be captured completely intact and undistorted by the sectioning required for traditional microscopy techniques.
This is the final version. Available on open access from Nature Research via the DOI in this record
Code availability:
The ESRF High Speed Tomography in Python (PyHST2) software which was used to reconstruct the phase images is open source and can be found at: https://software.pan-data.eu/software/74/pyhst2. The current pipeline for processing the raw data prior to its use in the PyHST2 algorithm is a large collection of scripts in MATLAB, Python and GNU Octave which makes it difficult to bundle into a single tomography pipeline. However, the ESRF is currently working to convert all of the scripts to Python to create a completely open source pipeline, though additional computing power, such a high performance computing cluster will likely be necessary. All additional image processing was done using open source Python libraries; these have been noted at the appropriate stages in the text.