Hannover 2016 – scientific programme
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Q: Fachverband Quantenoptik und Photonik
Q 32: Biophotonics
Q 32.2: Talk
Wednesday, March 2, 2016, 11:15–11:30, f342
Extending the applicability of Scanning Laser Optical Tomography using antibody staining and nonlinear contrast mechanisms — •Lena Nolte1, Nadine Tinne1, Georgios Antonopoulos1, Marko Heidrich1, Jennifer Schulze2, Kristin Schwanke2, Robert Zweigerdt2, Athanasia Warnecke2, Alexander Heisterkamp3, Tammo Ripken1, and Heiko Meyer1 — 1Laser Zentrum Hannover, Germany — 2Medizinische Hochschule Hannover, Germany — 3Leibniz Universität Hannover, Germany
Scanning laser optical tomography (SLOT) enables three dimensional visualization of large samples up to a magnitude of several centimeters using absorption and autofluorescence as intrinsic contrast mechanisms. However, this intrinsic contrast is sometimes not strong enough to image significant details inside the sample. One challenge is the visualization of hair cells and neurofilaments inside the human cochlea. For this reason, we developed a protocol for decalcification, antibody staining and optical clearing to image the cochlea in toto using SLOT.
Optical clearing is an efficient way to look into thick and turbid samples, but also prohibits the application of in vitro studies. Using near-infrared light, the scattering coefficient of the sample is lower and imaging of non-cleared samples can be improved. Therefore, we integrated a fs-pulsed laser source into the SLOT to enable the generation of two-photon fluorescence inside the sample. This way, living cell aggregates, with a diameter up to hundreds of micrometers, can be studied with respect to their three-dimensional structure without optical clearing.