Fig. 4
From: High-resolution spatiotemporal pHe and pO2 imaging in head and neck and oesophageal carcinoma cells
Spatial resolution of cell types based on extracellular acidification measured using STO2p-Q. a 7.5 × 105 cells (FaDu, Kyse30, Cal33, Cal27, HNEpC, HFF) were seeded into 4-well silicon inserts, as shown on the left. Heat maps represent pH after 3 and 20 min, measured using STO2p-Q. b Different cell numbers of FaDu, Kyse30, Cal33, and Cal27 were seeded into 4-well silicon μ-inserts (k, thousand, e.g. 6.1k = 6100 cells). Heat maps represent pH after 10 and 60 min. HFF and Kyse30 (c) or FaDu (d) cells expressing mCherry were seeded into a 6-well plate to create different 2D-landscapes. pH heat maps and microscopic images were overlaid, and the borders of cell layers outlined in white (Kyse30/FaDu) and blue (HFF) based on phase contrast images (BF) and mCherry signal. Heat maps represent pO2 after 2 and 20 min (c) or 3 and 20 min (d). Scale bars, 5 mm. d HFF and FaDu cells were seeded into a 6-well plate to create different 2D-landscapes. pO2 heat maps and microscopic images were overlaid, and the borders of cell layers outlined in white/black (FaDu) and blue (HFF) based on phase contrast (BF) and mCherry images. Heat maps represent pO2 after 3 and 20 min. Scale bars, 5 mm. HFF cells were seeded on Kyse30 (e) or FaDu (f) colonies expressing mCherry growing in 6-well plates. pH heat maps and microscopic images were overlaid, and the Kyse30/FaDu colonies outlined in black or white based on phase contrast images (BF, bight field) and mCherry signal. Heat maps represent pH after 5 and 15 min. Scale bars, 2.5 mm. Noise filter was applied with a smoothing factor of 2 for a to d. For all measurements, representative images are shown