A novel tape-free sample preparation method for human osteochondral cryosections for high throughput hyperspectral imaging

Xiwei Fan; Bogdan Donose; Michael W M Jones; Daryl Howard; Jari Torniainen; Karl Bertling; Xiao Guo; Cameron M Kewish; Kah Meng Lee; Antonia Rujia Sun; Aleksandar Rakic; Ross Crawford; Isaac O Afara; Indira Prasadam

doi:10.1007/s00418-024-02338-1

A novel tape-free sample preparation method for human osteochondral cryosections for high throughput hyperspectral imaging

Histochem Cell Biol. 2024 Dec 2;163(1):16. doi: 10.1007/s00418-024-02338-1.

Authors

Xiwei Fan^{1

2

3}, Bogdan Donose⁴, Michael W M Jones^{5

6}, Daryl Howard⁷, Jari Torniainen⁴, Karl Bertling⁴, Xiao Guo⁴, Cameron M Kewish^{7

8}, Kah Meng Lee⁵, Antonia Rujia Sun^{2

3}, Aleksandar Rakic⁴, Ross Crawford^{2

9}, Isaac O Afara^#^{10

11}, Indira Prasadam^#^{12

13}

Affiliations

¹ Department of Orthopaedic Surgery, The Second Xiangya Hospital of Central South University, Changsha, China.
² School of Mechanical, Medical & Process Engineering, Queensland University of Technology, 60 Musk Ave/Cnr. Blamey St, Kelvin Grove, Brisbane, QLD, 4059, Australia.
³ Centre for Biomedical Technologies, Queensland University of Technology, 60 Musk Ave/Cnr. Blamey St, Kelvin Grove, Brisbane, QLD, 4059, Australia.
⁴ School of Electrical Engineering and Computer Science, Faculty of Engineering, Architecture and Information Technology, University of Queensland, Brisbane, Australia.
⁵ Central Analytical Research Facility, Queensland University of Technology, Brisbane, Australia.
⁶ School of Chemistry and Physics, Queensland University of Technology, Brisbane, Australia.
⁷ Australian Nuclear Science and Technology Organisation, Australian Synchrotron, Melbourne, Australia.
⁸ Department of Chemistry and Physics, La Trobe University, Bundoora, Australia.
⁹ The Prince Charles Hospital, Brisbane, Australia.
¹⁰ School of Electrical Engineering and Computer Science, Faculty of Engineering, Architecture and Information Technology, University of Queensland, Brisbane, Australia. [email protected].
¹¹ Department of Technical Physics, University of Eastern Finland, Kuopio, Finland. [email protected].
¹² School of Mechanical, Medical & Process Engineering, Queensland University of Technology, 60 Musk Ave/Cnr. Blamey St, Kelvin Grove, Brisbane, QLD, 4059, Australia. [email protected].
¹³ Centre for Biomedical Technologies, Queensland University of Technology, 60 Musk Ave/Cnr. Blamey St, Kelvin Grove, Brisbane, QLD, 4059, Australia. [email protected].

^# Contributed equally.

PMID: 39621099
DOI: 10.1007/s00418-024-02338-1

Abstract

Understanding the osteochondral junction, where non-mineralised cartilage and mineralised bone converge, is crucial for joint health. Current sample preparation techniques are insufficient for detailed spatial hyperspectral imaging analysis. Using the enhanced Kawamoto method, we used the super cryo embedding medium's temperature-dependent properties to transfer high-quality tissue samples onto slides for spatial imaging analysis. We transferred osteochondral samples using a tape-free system and successfully tested them in hematoxylin and eosin (HE), Safranin-O, nanomechanical assessments and nano-Fourier transform infrared (FTIR) mapping. This protocol elucidates the structural and elemental gradients, mechanical characteristics and distinctive biochemical layering, making it a useful tool for analysing biochemical properties' co-distribution in healthy and diseased situations.

Keywords: Cryosectioning; Molecular imaging; Nano-mechanics; Osteochondral unit; Trace element; X-ray ptychography.

MeSH terms

Cartilage / chemistry
Cartilage / metabolism
Cryoultramicrotomy
High-Throughput Screening Assays
Humans
Hyperspectral Imaging*
Spectroscopy, Fourier Transform Infrared

Abstract

MeSH terms

Grants and funding