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. 2022 Mar 2;414(18):5483–5492. doi: 10.1007/s00216-022-03944-5

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — (ai–aiii) Pre-subcellular biopsy images of skeletal muscle fibres, corresponding to BF (ai), myonuclei stained with DAPI (aii), and IF staining of MTCOI (aiii). (bi–biii) Post-biopsy images, again BF (bi), DAPI (bii), and MTCOI IF staining (biii). Evidence of successful biopsies is shown in BF (bi), but a reduction in MTCOI fluorescence was not shown post-biopsy (biii). The relative position of the biopsy to myonuclei is shown relative to DAPI staining, which indicates that biopsies taken are from the intermyofibrillar region and not from the perinuclear region. The biopsy sites in all images (pre- and post-biopsy) were delineated within an orange box. A topographical scan of the area shown within the orange box (32 × 32 µm) further supported the successful biopsy as shown by an indentation in the skeletal muscle fibre, measuring approx. 7 µm in diameter, consistent with the biopsy site observed in BF (c)