Spectrally resolved multiphoton microscopy for the identification of biomarkers (Conference Presentation)

Björn-Ole Meyer; Dominik Marti; Peter E. Andersen

doi:10.1117/12.2508311

4 March 2019 Spectrally resolved multiphoton microscopy for the identification of biomarkers (Conference Presentation)

Björn-Ole Meyer, Dominik Marti, Peter E. Andersen

Author Affiliations +

Proceedings Volume 10882, Multiphoton Microscopy in the Biomedical Sciences XIX; 1088214 (2019) https://doi.org/10.1117/12.2508311
Event: SPIE BiOS, 2019, San Francisco, California, United States

Abstract

We have developed a system capable of resolving Two-Photon Fluorescence Emission (TPFE) and Second Harmonic Generation (SHG) signals with high spectral resolution for the characterization of biomarkers. In Multiphoton Microscopy, those biomarkers are TPFE and SHG signals that carry valuable information on morphological and functional biological features, such as the presence of Green Fluorescent Protein (GFP) in a Zebrafish during the building of organs, or the ratio of pyridine nucleotide (NAD(P)H) and flavin adenine dinucleotide (FAD) in the classification of cancerous tissue. For this purpose, separation of different signals into channels is typically achieved by the use of optical filters. In doing so, signal spectra can be unknown or overlapping, creating a crosstalk in between the channels. Previously the choice of such filters relied on prior knowledge or heuristic testing. Our system allows for the study of biomarkers due to spectrally resolved imaging. It therefore enables the appropriate selection of channels, tailored to the application, when building fast diagnostic systems. Additionally, knowledge of the spectra can be used to avoid the crosstalk in between channels or compensate for it computationally. To demonstrate the capabilities of the system, we recorded spectrally resolved images of tissue and cell samples. Structural and functional biological features were identified and their spectra could be evaluated. Thus, appropriate filter setups for diagnostic imaging can be suggested and confirmed by means of integration over defined virtual channels.

Conference Presentation

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Björn-Ole Meyer, Dominik Marti, and Peter E. Andersen "Spectrally resolved multiphoton microscopy for the identification of biomarkers (Conference Presentation)", Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 1088214 (4 March 2019); https://doi.org/10.1117/12.2508311

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KEYWORDS

Multiphoton microscopy

Diagnostics

Green fluorescent protein

Imaging systems

Second-harmonic generation

Tissues

Harmonic generation

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