Prime 10

An Olympus BX51WI upright microscope and a water-immersion LUMPlan FL/IR 20×/0.50 W objective were used. Excitation (740 nm) was provided by a Prairie View Ultima multiphoton laser scan unit powered by a Millennia Prime 10 W diode laser source pumping a Tsunami Ti: sapphire laser (Spectra-Physics, Mountain View, CA, USA). Blood plasma was labeled by i.v. tetramethylrhodamine isothiocyanate dextran (155 kDa) in physiological saline (5 % wt/ vol). All microvessels in an imaging volume (500 × 500 × 300 μm) were scanned at each study point, measuring the diameter and blood flow velocity in each vessel (3–20 μm Ø). Tetramethylrhodamine fluorescence was band pass filtered at 560–600 nm and NADH autofluorescence at 425–475 nm. Imaging data processing and analysis were carried out using the NIH ImageJ.

An Olympus BX51WI upright microscope and a water-immersion LUMPlan FL/IR 20×/0.50 W objective were used. Excitation (740 nm) was provided by a Prairie View Ultima multiphoton laser scan unit powered by a Millennia Prime 10 W diode laser source pumping a Tsunami Ti: sapphire laser (Spectra-Physics, Mountain View, CA, USA). Blood plasma was labeled by i.v. injection of tetramethylrhodamine isothiocyanate dextran (155 kDa) in physiological saline (5 % wt/ vol). All microvessels in an imaging volume (500 × 500 × 300 μm) were scanned at each study point, measuring the diameter and blood flow velocity in each vessel (3–20 μm Ø). Tetramethylrhodamine fluorescence was band pass filtered at 560–600 nm and NADH autofluorescence at 425–475 nm. Imaging data processing and analysis were carried out using the NIH ImageJ processing package.