Uplanfln 40 1.30 oil objective

Microscopy was performed using an Olympus BX61 VS120-S5 Virtual Slide Scanning System with UPlanSApo 40×/0.95 objective, Olympus XM10 monochrome camera, and Olympus VS-ASW FL 2.7 imaging software. Additionally, 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. Live cell culture imaging was performed using an Olympus IX81 with LUCPlan FLN 20×/0.45 objective, Olympus XM10 monochrome camera and Olympus CellSens 1.13 software. Analysis and quantification were performed using Olympus CellSens 1.13, MetaMorph Premier 7.7.0.0, Adobe Photoshop CS6, and ImageJ software.

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.