TWO-PHOTON IMAGING OF NAD(P)H IN TUMOR SPHEROIDS

Project: Research project

Project Details

Description

We propose to use two-photon laser scanning microscopy (TPLSM) to measure
NADH and NADPH (NAD(P)H) fluorescence in the multicell tumor spheroid as an
indication of the development of anoxia during PDT, a novel laser-based
cancer therapy which is currently undergoing clinical trials worldwide. To
demonstrate that TPLSM of NAD(P)H can be used to monitor anoxia in the
multicell tumor spheroid, it will be necessary to demonstrate that NAD(P)H
fluorescence can be correlated to cellular anoxia ina the subpopulations of
cells which comprise a spheroid. In Aim 1, we will measure athe
fluorescence of normal and EMTS/Ro cell monolayers equilibrated at various
oxygen partial pressures. Additionally, the fraction of fluorescence
originating from the mitochondria and the fraction of fluorescence which is
dependent on the oxygen partial pressure will be determined. Multicell
tumor spheroids equilibratfed at various oxygen partial pressures will then
be used to establish and control the extent of anoxia in this model tumor
system. In Aim 2, TPLSM will be correlated to anoxia in the spheroid as
measured with an oxygen sensitive microelectrode with a 5 mum tip diameter
using information obtained from Aim 1 to determine the extent of the anoxic
region from NAD(P)H fluorescence images. Studies conducted as part of Aim
3 will then monitor the onset and progression of anoxia during FDT
conducted at several optimal fluence raises known to cause anomia of
various extent. Finally, Aim 4 will explore potential analytical methods
to utilize NAD(P)H fluorescence as a direct non-invasive diagnostic
indicator of therapeutic efficacy in this model tumor system in vitro as
well as tumors in vivo. The development of this technique would be
extremely useful in treatment planning and monitoring response to treatment
interventions such as radiation and photodynamic therapy of cancer.
StatusFinished
Effective start/end date8/5/968/4/99

Funding

  • National Cancer Institute
  • National Cancer Institute
  • National Cancer Institute

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