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In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue

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Abstract

High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

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Acknowledgments

Authors Karsten König and Martin Weinigel are empoyees of the Jenlab company, which produces the multiphoton microscope used in this study. However no financial support was received from this company and the company had no influence on the experiments performed. The Friedhelm-Frees-Foundation (http://www.klinik.uni-mainz.de/friedhelmfreesstiftung/) supported this study with a Grant of 10,000 Euro.

Author information

Authors and Affiliations

  1. University Medical Centre, Johannes Gutenberg-University, Langenbeckstraße 1, 55101, Mainz, Germany

    Sven R. Kantelhardt, Darius Kalasauskas, Ella Kim & Alf Giese

  2. Department for Biophotonics and Laser Technology, Saarland University, Campus A5.1, 66123, Saarbrücken, Germany

    Karsten König & Aisada Uchugonova

  3. JenLab GmbH, Schillerstrasse 1, 07745, Jena, Germany

    Karsten König & Martin Weinigel

Authors
  1. Sven R. Kantelhardt
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  2. Darius Kalasauskas
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  3. Karsten König
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  4. Ella Kim
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  5. Martin Weinigel
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  6. Aisada Uchugonova
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  7. Alf Giese
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Corresponding author

Correspondence to Sven R. Kantelhardt.

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Conflict of interest

Authors professor Dr. Karsten König and Dr. Martin Weinigel work with the JenLab GmbH, that produces the MPT used in this study. Apart from this the authors declare that they have no conflict of interest.

Additional information

This work is part of the doctoral thesis of Darius Kalasauskas.

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Supplementary material 1 (PPT 5798 kb)

Supplementary material 2 (PPT 1860 kb)

Supplementary fig. 1—The figure shows the exact locations where the specimen shown in figs. 2A, B were taken from. A corresponds to the surface of the murine cortex, whereas B is taken from a deeper portion within the murine cortex (luxol fast blue stain (LFB) identifying myelin fibers, the white bar on the inlets A and B corresponds to a distance of 50 µm).

Supplementary fig. 2—Panel A shows a human transitional meningioma WHO°I and B brain metastasis of an undifferentiated adenocarcinoma of a breast cancer. The left column shows fluorescence intensity images (images marked with a1, b1; excitation 760nm), while second column (a2, b2) shows the corresponding FLIM images using a continuous color-coding (red stands for a short fluorescent lifetime of 500 ps and blue for long fluorescent lifetimes of up to 3000ps). The white bars correspond to a distance of 50 µm. The right column (a3, b3) shows the corresponding conventional histology (HE).

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Kantelhardt, S.R., Kalasauskas, D., König, K. et al. In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue. J Neurooncol 127, 473–482 (2016). https://doi.org/10.1007/s11060-016-2062-8

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  • DOI: https://doi.org/10.1007/s11060-016-2062-8

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