Fig. 3: Intravital short-term live imaging and long-term cell tracing of the posterior midgut. | Communications Biology

Fig. 3: Intravital short-term live imaging and long-term cell tracing of the posterior midgut.

From: Intravital imaging strategy FlyVAB reveals the dependence of Drosophila enteroblast differentiation on the local physiology

Fig. 3

a Schematic illustration of the adult Drosophila midgut. The epithelium mainly consists of four types of cells: intestinal stem cells (ISCs), enteroendocrine cells (EEs), enteroblasts (EBs), and enterocytes (ECs). ISCs and EBs were both labelled by esg-Gal4, and they were distinguished by the presence of NRE-GFP in the EBs. EEs were labelled by pros-Gal4, pros-QF2, and Rab3-Gal4. ECs were identified by the polyploid nuclei. b Visualization of the ISC lineage cells using FlyVAB. ISCs (tdTomato+, NRE-GFP-), EBs (tdTomato+, NRE-GFP+), EEs (mcherry+) and ECs (H2B-RFP, polyploid nuclei) visible on a laser scanning confocal microscope or widefield fluorescence microscope. c Determination of the visualized region in the midgut by photobleaching. The midgut was barely visible on the FlyVAB device after photobleaching. After dissection and fixation, the bleached region (BR, solid line) was located in the posterior midgut (PMG, dashed line). Genotype: esg-Gal4 10×UAS-myr:tdTomato. See also Supplementary Fig. 4. d Time-lapse imaging of calcium signal. e A total of 354 time points was acquired in 10 min. See also Supplementary Movie 3. f Serial imaging of the same region re-identified by a pattern of 7 EEs over 10 days. A pair of newly formed EEs (the dashed square is enlarged in the lower right corner) was identified. Scale bars are 10 μm in b, d, and e, 100 μm in c.

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