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. 2021 Feb 26;10(3):499. doi: 10.3390/cells10030499

Figure 7.

Figure 7

Intravital bioluminescence and fluorescence microscopy of pancreas window chamber. Diagram with measurements and representative photograph of (a) pancreas window chamber and (b) imaging stage insert. (c) Intravital microscopy images of the pancreas using wild type C57BL/6 abdominal window chamber-bearing animals with Pan02 NF-κB-FLuc Dendra2 pancreatic tumors. Representative intravital image of NF-κB-FLuc bioluminescence and Dendra2 fluorescence of Pan02 pancreatic tumor progression over time at 10X objective (BLI, 10 min acquisition, open filter; Dendra2 green, GFP cube, 200 ms exposure; Dendra2 red, DsRed cube, 500 ms; scale bar represents 100 μm). (d) Representative fluorescence image of Pan02 tumor pre- and post-photoswitching (DAPI cube, 10 min exposure) at 10X objective (Dendra2 green, GFP cube, 200 ms exposure; Dendra2 red, DsRed cube, 500 ms; scale bar represents 100 μm). Quantification of NF-κB-FLuc bioluminescence counts of (e) total tumor and (f) photoconverted area of the tumor pre- and 24 h post-Dendra2 photoconversion (data are represented as mean ± SEM, n = 3 animals). (g) Representative intravital images of resolved bioluminescence of L-012 innate immune cell activity and Pan02 NF-κB-FLuc reporters (2X objective, sequential imaging of L-012 (20 min acquisition, open filter) and NF-κB-FLuc (10 min acquisition, open filter)), overlaid image of bioluminescent reporters (combined; pseudocolored red), and tumor Dendra2 (Dendra2 green, GFP cube, 2 s exposure); scale bar represents 1000 µm. (h) Representative intravital images of abdominal window chamber implanted at week 14 visualizing the pancreas of KPC-Luc animals at 2X (FLuc, 2 min acquisition, open filter; scale bar represents 1000 µm) and 10X (FLuc, 2 min acquisition, open filter; scale bar represents 100 µm); n = 2 animals. Pancreas tumor growth monitored by FLuc bioluminescence following D-luciferin injection. Vasculature imaged post i.v. injection of dextran-FITC (200,000 MW, 4 mg/mL, epifluorescence: GFP cube; 2X, 5 s exposure; 10X, 2 s exposure).