Artikel
Multiphoton excited fluorescent microscopy detection of residual tumour and invasive glioma cells
Multiphotonen angeregte Fluoreszenzmikroskopie zur Darstellung von Resttumor und invasiven Gliomzellen
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Autoren
Veröffentlicht: | 4. Mai 2005 |
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Gliederung
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Objective
Upcoming optical technologies for non-invasive analysis of normal brain and brain tumour tissue offer novel perspectives for the intraoperative detection of residual tumour. Multiphoton excited in vivo fluorescent microscopy is a laser-based technology that allows subcellular resolution of native tissues in situ. Using experimental gliomas and specimens from human brain tumours, we have evaluated the potential of this technology as an adjunct to controlling the extent of resection.
Methods
Invasive and non-invasive experimental gliomas were generated in nude mice and ex vivo specimens were analysed by multiphoton microscopy. Corresponding tissue blocks were fixed and processed for conventional histology. Biopsies of human brain tumours were obtained during resection of glial tumours and biopsy sites were documented using neuronavigation. The native tissue blocks were analyzed by multiphoton microscopy and the microanatomy of specimens was correlated with MRI findings and conventional histology.
Results
Multiphoton excited fluorescent microscopy of ex vivo brain tumour specimens demonstrated visualisation of the cellular composition of solid tumour allowing the discrimination of individual tumour cells, tumour cell clusters and vasculature. At the cellular level a clear distinction of elements of normal brain and invasive protrusions of the tumour or single invading cells could be made. Subcellular structures of tumour cells such as nucleoli could be visualised up to 100 µm into solid tissue. In addition to visualisation of tumour anatomy laser excitation demonstrated cellular clusters within tumour areas that showed brisk fluorescence. A similar phenomenon was observed in individual invasive cells. These data suggest that distinct areas of a tumour contain cells producing excitable endogenous biomolecules that can be identified by specific laser wave lengths.
Conclusions
Optical technologies allow non-contact/non-invasive analysis of native tissues reaching a sub micron resolution. Multiphoton microscopy allows a discrimination of different cell types, neurons, glia, or tumour cells and visualisation of organelles. In addition, selective excitation of endogenous biomolecules offers means of imaging cellular metabolism or cellular function. Optical technologies integrated into neurosurgical equipment may potentially provide a structural and possibly functional analysis of the resection margins during surgery for glial tumours.