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

Figure 2.

Figure 2

Intravital dorsal skin window chamber bioluminescence and fluorescence imaging. (a) Modified skin window chamber implant where front plate flange was shorted to 20 mm (top photos) and custom imaging stage insert (bottom photos; 11 cm diameter insert with a 5.5 cm × 3 cm cutout for the animal to lay flat). A pin protruding perpendicular to the custom imaging stage insert (red arrow) fits into the window chamber implant (blue arrow) to further lock the animal in place during imaging. (b) Representative bioluminescence images of B16F10 κB5-FLuc GFP tumors in skin window chamber-bearing animals using the macro-imaging system (macro, scale bar represents 1000 µm) and the microscopy system at 2X (5 min acquisition, open filter, scale bar represents 1000 µm) and 20X (10 min acquisition, open filter, scale bar represents 100 µm) where the red circle on the 2X image indicates a magnified area. (c) Linearity analysis between the macro-imaging system (IVIS) and micro-imaging system (Nikon); the correlation was measured at 1 (p < 0.0001) using n = 5 images where data were expressed as mean ± s.e.m. (error bars fall within the data point). Representative in vivo microscopic imaging over time of B16F10 κB5-FLuc GFP reporter tumor growth wherein GFP monitors B16F10 tumor growth and BLI monitors FLuc bioluminescence indicative of transcriptional activation of NF-κB at (d) 2X magnification (GFP cube, 500 ms; BLI, 5 min acquisition, open filter, scale bar represents 1000 μm, red box is corresponding area imaged at 20X) and (e) 20X magnification (GFP cube, 20 ms; BLI, 10 min acquisition, open filter, scale bar represents 100 μm).