Axioplan 2ie microscope

Full-scale images of the samples before and after tests were acquired with a Dino-Lite^® Edge AM7915MZTL microscope (AnMo Electronics Corporation, Taipei, Taiwan) with an Open Cap N3C-O (9.5 mm length), varying Dino-Lite^® lighting levels (Flexible LED Control) that were controlled through the DinoCapture 2.0 software (Almere, The Netherlands), at a magnification of approximately ×20 (scale is 2 mm).
Detailed images of the samples’ surfaces were acquired using an Axioplan 2ie microscope (Zeiss, Germany), equipped with an incident halogen light illuminator (tungsten light source, HAL 100) and coupled with a DXM1200F digital camera and ACT-1 control software (Nikon, Japan). Micrographs were captured within a few days of exposure to CO₂ and again after 35 weeks, using reflected (incident) light in brightfield, darkfield, and cross-polarized modes, and transmitted light in cross-polarized mode, at varied magnifications (×50, ×100, ×200, and ×500).

Throughout the paper (except in Figure 2F–G’’’ and Figure 2—figure supplement 2A–E), live tissues were used for imaging. Live imaging of the Drosophila imaginal tissues was performed as previously described (Xu et al., 2019 (link)). Briefly, freshly dissected wing or eye imaginal discs from third instar larvae were pipetted into a ~40 μl droplet of Schneider’s Drosophila Medium supplemented with 10% fetal bovine serum and mounted on a glass slide. To support the tissue, spherical glass beads (Cospheric, Product ID: SLGMS-2.5) of ~50 μm in diameter were placed under the cover slip. The mounted samples were immediately imaged on Zeiss LSM 880 or LSM 800 confocal microscopes. Throughout the paper, apical tissue views were shown as maximum projections of the most apical optical sections (0.75 μm/section, four to five sections) generated using Image J; for basal views, single sections ~10.5 μm below the apical surface were shown. Widefield fluorescence imaging of live wing imaginal discs was done using a Zeiss Axioplan 2ie microscope with an Orca ER camera and Zeiss AxioVision software.