[abstract] IMAGING LIVING NEURONS (IN VITRO) USING HYPERBARIC MICROSCOPY METHODS.

Rubicon Research Repository/Manakin Repository

[abstract] IMAGING LIVING NEURONS (IN VITRO) USING HYPERBARIC MICROSCOPY METHODS.

Show full item record


Title: [abstract] IMAGING LIVING NEURONS (IN VITRO) USING HYPERBARIC MICROSCOPY METHODS.
Author: Dean, JB; D’Agostino, DP
Abstract: INTRODUCTION: The limiting factor in conducting cellular physiological studies under hyperbaric conditions is the requirement to adapt cutting edge research tools for use inside a sealed pressure vessel. To date, we have successfully adapted electrophysiology and microscopy equipment for research on cellular mechanisms of CNS oxygen toxicity. This presentation summarizes our recent revisions to our hyperbaric microscope, which have improved its ease of use for in vitro experiments in rat brain tissue slices during helium compression and decompression. MATERIALS AND METHODS: Optical components, cameras, light guides, manipulators, breadboards and miscellaneous components were purchased primarily from Nikon, Roper Scientific, and Edmund Optics and assembled together as a custom-built, fixed-stage upright microscope. The microscope and its supporting equipment fit inside a refurbished Bethlehem hyperbaric chamber (Reimers Systems Inc., Hyperbaric Clearing House) measuring 18” x 37” with a MWP= 150 psig. Three moveable equipment sleds support the microscope, tissue slice bath, and micromanipulators. RESULTS: The integrated microscopy system has been used successfully at ≤6 ATA helium. The microscope has multiple applications, including the following: low-magnification transmitted illumination and low angle illumination for pseudo-darkfield imaging, both of which aid the investigator in precise placement of various recording probes (≤3) into the brain slice; low- and high-magnification imaging of neurons in slices labeled with fluorogenic probes that measuring intracellular levels of superoxide, nitric oxide, and pH in real time; and IR-video imaging with Hoffman Modulation Contrast optics for visually selecting individual neurons for electrophysiological recording. Currently, the microscope is being used to study the effects of HBO2 on superoxide production in neurons. CONCLUSIONS: It is possible to conduct imaging experiments that require high-power magnification under hyperbaric conditions. This novel research tool has proven useful for ONR-supported studies of the cellular mechanisms of CNS oxygen toxicity and other baro-related problems. (ONR N000140710890; DoD-ONR DURIP N000140210643)
Description: Abstract of the Undersea & Hyperbaric Medical Society 2008 Annual Scientific Meeting June 26-28, 2008 Salt Lake City Marriott Downtown, Salt Lake City, Utah.
URI: http://archive.rubicon-foundation.org/7890
Date: 2008

Files in this item

Files Size Format View
abstract.txt 193bytes Text file View/Open

This item appears in the following Collection(s)

  • UHMS Meeting Abstracts
    This is a collection of the published abstracts from the Undersea and Hyperbaric Medical Society (UHMS) annual meetings.

Show full item record

Browse

My Account