Laser Scanning Multiphoton Microscopes
Future developments will include exploring performance
improvements for even higher imaging resolution and depth by considering adaptive optics techniques. Further improvements in computing power and intelligent algorithms will provide the workflow necessary for high-throughput application such as digital (imaging) pathology. Te modular platform allows reasonably straightforward additions for expansion with other optical modalities to enhance the utility.
Acknowledgements We are thankful for support and feedback from the
following collaborators: Frank Maigler, Stella Gänger and Prof. Katharina Zimmermann, HBC Biberach; Bettina Sailer, Tomas Kellerer, and Prof. Oliver Hayden, Transla- TUM - TUM, Munich; Amelie Erben and Dr. Stefanie Sud- hop, Center for Applied Tissue Engineering and Regenerative Medicine (CANTER), HM, Munich; Prof. Christophe Lamy, LAMYLAB - University of Geneva; Dr. Kim Ferrari and Prof. Bruno Weber, Neuroscience Center UZH-Zurich; Dr. William Lackington, Empa, St Gallen; and Spirochrome Ltd, Stein am Rhein, for providing samples, images, and dedicating time and resources to the experiments mentioned.
Figure 7: Multiphoton maximum intensity projections of 3D z-stacks of tissue- engineered constructs. A) Two-photon fluorescence of a 3D nano-printed artifi- cial lung tissue composed of GM10-RoseBengal (green) seeded with A549 lung cells and stained for actin (red) and the cell nuclei (blue). Engineered by Amelie Erben, supervised by Dr. Stefanie Sudhop at the Center for Tissue Engineering, Munich. B–D) Combination of label-free SHG images originating from collagen ((magenta) (B), blue (C, D)) and two-photon florescence images of stained cells grown in the collagen scaffold. B) Primary bone cells stained for actin (yellow) and the cell nuclei (blue). C) Primary bone cells stained for actin (red) and fibrino- gen (yellow). D) Top view of a label-free collagen scaffold, SHG signal (blue). Engineered by Dr. William Lackington, Empa, St. Gallen. Scale bars = 50 μm.
complementary microscopy diagnostics. In this paper we dem- onstrated SHG, widefield, two-photon, and FLIM imaging together as applied to applications in tissue engineering, drug delivery, neuroscience, and cancer research. In addition, we showed multiple setup configurations, including live-animal and upright arrangements, and imaged many different sample types. Despite the enormous flexibility, the microscope deliv- ers excellent performance, mechanical stability, and the optical performance necessary for quality fast-imaging.
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