Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching.

Abstract:

:Protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (5-ALA) is increasingly used as a fluorescent marker for fluorescence-guided resection of malignant gliomas. Understanding how the properties of the excitation light source and PpIX fluorescence interact with the surgical microscope is critical for effective use of the fluorescence-guided tumor resection technique. In this study, we performed a detailed assessment of the intensity of the emitted blue light and white light and the light beam profile of clinical grade operating microscopes used for PpIX visualization. These measurements revealed both recognized fluorescence photobleaching limitations and unrecognized limitations that may alter quantitative observations of PpIX fluorescence obtained with the operating microscope with potential impact on research and clinical uses. We also evaluated the optical properties of a photostable fluorescent standard with an excitation-emission profile similar to PpIX. In addition, we measured the time-dependent dynamics of 5-ALA-induced PpIX fluorescence in an animal glioma model. Finally, we developed a ratiometric method for quantification of the PpIX fluorescence that uses the photostable fluorescent standard to normalize PpIX fluorescence intensity. This method increases accuracy and allows reproducible and direct comparability of the measurements from multiple samples.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Belykh E,Miller EJ,Patel AA,Bozkurt B,Yağmurlu K,Robinson TR,Nakaji P,Spetzler RF,Lawton MT,Nelson LY,Seibel EJ,Preul MC

doi

10.1038/s41598-018-30247-6

subject

Has Abstract

pub_date

2018-08-22 00:00:00

pages

12543

issue

1

issn

2045-2322

pii

10.1038/s41598-018-30247-6

journal_volume

8

pub_type

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