Xm10 monochrome camera

Microscopy was performed using an Olympus BX61 VS120-S5 Virtual Slide Scanning System with UPlanSApo 40×/0.95 objective, Olympus XM10 monochrome camera or Allied vision Pike F-505C color camera, and Olympus VS-ASW FL 2.7 imaging software. Confocal images were acquired using an Olympus FV1000 Confocal Microscope with UPlanFLN 40×/1.30 oil objective and Olympus FV-ASW version 4.2 imaging software. Perilipin-1 quantification was performed by thresholding the image to exclude non-specific staining and then calculating the area of each lipid deposit encircled by perilipin-1 (excluding those in the epi- or perimysium) using MetaMorph software (Molecular Devices). The total area encircled by perilipin-1 was calculated for all lipid deposits across the muscle and expressed relative to the total cross-sectional area. PDGFRα and AnxA2 positive area was calculated using CellSens software (Olympus) by thresholding to remove non-specific staining and calculating the total positive area (again, excluding any epi- or perimysial staining) relative to the entire muscle cross-section.

For immunhistochemical staining of HAP1 cells, culture media was aspirated and cells were washed once with 1× PBS. Cells were fixed with ice-cold 4% paraformaldehyde in 1× PBS containing 15% sucrose for 20 min at room temperature (RT) and subsequently permeabilized with 0.1% Triton X-100 in 1× PBS for 3 min at 4 °C. For F-actin staining, Phalloidin-iFluor 594 (Abcam, Cambridge, United Kingdom) was diluted 1 : 1000 in 1% bovine serum albumin (Carl Roth, Karlsruhe, Germany) in 1× PBS and for staining of cell nuclei 0.5 μg mL⁻¹ DAPI (Carl Roth) was added. Cells were incubated for 1 h at RT, washed with 1× PBS and coverslips were mounted on microscope slides with Shandon Immu-Mount mounting media (Thermo Fisher Scientific).
Bright-field images of printed HAP1 and HEK293H cells were captured with a CKX53 inverted microscope with integrated Phase Contrast (iPC) and a XM10 monochrome camera (Olympus, Shinjuku, Japan). Fluorescence images were acquired with an IX83 inverted imaging system with a DP80 camera (Olympus) and a 4-channel high-specification LED System (Judges Scientific, London, United Kingdom). Olympus cellSense software was used with both microscopes, and adjustments of brightness and contrast were carried out with ImageJ (NIH, Bethesda, MD, USA).

Images were acquired on the Olympus BX61 VS120‐S5 Virtual Slide Scanning System Microscope with UPlanSApo 20x/0.75 and 40x/0.95 objectives, Olympus XM10 monochrome camera, and Olympus VS‐ASW FL 2.7 imaging software. Analysis was performed using Olympus CellSens 1.13 and FIJI ImageJ software.