100 w highpressure mercury burner

The PDLCs were characterized by brightfield and fluorescence microscopy with either a BX50 Upright Microscope (ACH 60X/0.80 ∞/0.17 objective) or an IX70 Inverted Microscope (ACH 60X/0.80 ∞/0.17 objective) with a 100 W High-Pressure Mercury Burner, Olympus (Waltham, MA, USA). Diluted samples of PDLCs were pipetted onto 25 × 75 × 1 mm microscope slides and placed under a glass coverslip (Fisher Scientific, Waltham, MA, USA) in preparation for imaging. The appropriate filters were used to separate DiO and DiD fluorescence. Microscopy pictures were acquired by a Rolera Bolt CMOS QImaging Camera (Surrey, BC, Canada). Images for both the PDLCs and the DBCO blocked negative control groups were acquired from three independent samples (n = 3) using ten images per sample.

Vessel formation was assessed at the aforementioned time points. Fluorescent images were captured utilizing an Olympus IX81 equipped with Disc Spinning Unit and a 100 W high-pressure mercury burner (Olympus America, Center Valley, PA), a Hamamatsu Orca II CCD camera (Hamamatsu Photonics, K.K., Hamamatsu City, Japan), and Metamorph Premier software (Molecular Devices, Sunnyvale, CA). Imaged beads were chosen at random provided that vessels emanating from a given bead did not form anastomoses with vessels from adjacent beads. Images from at least 30 beads per condition were captured over three separate trials at low magnification (4×) for each independent experiment and processed using the Angiogenesis Tube Formation module in Metamorph Premier (Molecular Devices). Each image was segmented and analyzed based on any tube-like pattern that falls within a specified minimum and maximum width of each segment above a contrast threshold. The total network length, the number of branch points, and number of segments were quantified.