Journal: Microvascular research

Article Title: A novel method for measuring hydraulic conductivity at the human blood-nerve barrier in vitro

doi: 10.1016/j.mvr.2016.08.005

Figure Lengend Snippet: Photographs of the hydraulic conductivity system, primarily consisting of a diffusion chamber connected to hydrostatic pressure reservoir, circulating water bath and bubble tracker apparatus utilizing digital time-lapse photography are shown. Figure 1a shows the experimental set-up without tubing connecting the hydrostatic reservoirs to the diffusion chambers. Figure 1b shows the set-up with the water bath disconnected and tubing from the hydrostatic pressure reservoir attached, while Figure 1c has the light box removed to clearly demonstrate the slotted plate containing precision bore glass tubes used to detect bubble displacement. Figure 1d is a flow diagram depicting the movement of medium through the hydraulic conductivity system with Figure 1e illustrating the modified EasyMount Ussing diffusion chamber with sealed fill ports (X). Black arrows depict the movement of medium into the chamber from the hydrostatic pressure reservoir and out to the glass tubing of the bubble tracker device, while white arrows indicate water flux across the Snapwell insert membranes with cultured confluent cells. Figure 1f is a schematic representation of the assembled bubble tracker device with bubbles (white circles) inserted into glass tubes and camera suspended overhead to record bubble displacement with a light box to illuminate the slotted plate and eliminate external light. Black arrows indicate fluid movement. Key: (1) adjustable height, 6-channel, hydrostatic pressure reservoir with 10 cc syringes, (2) aluminum breadboard, (3) 6-channel EasyMount stand, (4) EasyMount Ussing diffusion chamber with Snapwell inserts, (5) circulating, heated water bath, (6) slotted plate for holding 6 precision bore glass tubes, (7) light box, (8) camera mount, and (9) digital camera.

Article Snippet: 12 mm Snapwell cell culture inserts with 0.4 µm pore polyester membranes (Corning Costar; Kennebunk, ME) were placed in modified EasyMount Ussing diffusion chambers housed in a 6-Channel EasyMount stand (Physiologic Instruments, Inc.; San Diego, CA). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window caption a7 caption a8 Hydraulic conductivity measurement apparatus Photographs of the hydraulic conductivity system, primarily consisting of a diffusion chamber connected to hydrostatic pressure reservoir, circulating water bath and bubble tracker apparatus utilizing digital time-lapse photography are shown.

Techniques: Diffusion-based Assay, Modification, Cell Culture