Fig. 3: Quantification of intravascular and interstitial calcium concentration in the bone marrow.
a Merged Rhod-5N, AF488 and SHG signals and ratiometric imaging in the BM, demonstrated by a single image plane from a z-stack. Rhod-5N and AF488 as cell-impermeable dyes labeled vasculature and were sequestered in the interstitial space. The BM cells thus appeared as dark objects. b Calcium concentration calibration curves at pH = 7.0 and pH = 7.4 were obtained in vitro to convert the measured ratios to absolute calcium concentration (N = 3 independent experiments). The dashed black line is a fitting curve for the two data sets together. c Quantifications of intravascular and interstitial Rhod-5N/AF488 ratios in vivo as well as serum Rhod-5N/AF488 ratios measured in vitro (n = 25 BM cavities, N = 10 mice, N = 8 in vitro samples). d Corresponding [Ca2+] converted from Rhod-5N/AF488 rations in c using the calcium calibration curve (p = 0.001 between vessels and interstitium). e [Ca2+]e near arterioles/sinusoids (n = 8 BM cavities) and near endosteum (n = 25 BM cavities). c–e Two-sided Mann–Whitney test. Each data point represents a subregion from a BM cavity. Box and whiskers represent the median, 25 and 75 percentiles, and the 10–90% data range. Source data are provided as a Source Data file.
