Freely tunable spectral detection for multiphoton microscopy

Hilmar Gugel; Ingo Böhm; Felix Neugart

doi:10.1117/12.2509641

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Abstract

Multiphoton microscopy enables imaging deep inside living specimens. High photon flux is required for the nonlinear excitation of fluorescent markers, which confines the excitation to the small volume of the focal spot. This results in intrinsic optical sectioning and enables non-descanned detection of the fluorescence signal. Although tunable spectral detection has been standard in confocal microscopy for many years, it is still common in multiphoton microscopy to manually change the filters in the detection beam path of the microscope to accommodate for different fluorescence markers. We revolutionized the non-descanned detection by implementing a freely tunable spectral detector for four detection channels. This enables the adaptation to new transgenic markers in seconds and separates strongly overlapping spectra without mathematical restoration. Here, we introduce the technological concept and show its application in imaging of biological specimens demonstrating the capability of the spectral detector.

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Hilmar Gugel, Ingo Böhm, and Felix Neugart "Freely tunable spectral detection for multiphoton microscopy", Proc. SPIE 10882, Multiphoton Microscopy in the Biomedical Sciences XIX, 108820S (25 February 2019); doi: 10.1117/12.2509641; https://doi.org/10.1117/12.2509641

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