title: Real-time tissue offset correction system for intravital multiphoton microscopy.
abstract: The development of multi-photon intravital microscopy, in particular two-photon microscopy (2PM), has been a breakthrough technique for deep-tissue imaging of dynamic cell behavior inside live organisms and has substantially advanced the field of immunology. However, intravital time-lapse imaging over prolonged time periods is complicated by slow tissue drifts caused by vital activity, leading to shifting fields of views and making the acquired image sequence partially or completely unanalyzable. To solve this issue, we have established a system that performs continuous drift offset correction in real time using fine pattern matching during 2PM acquisition. We incorporated an extensive use of graphical processing unit (GPU) for high-speed computing required for real time correction during data acquisition. This allowed us to perform prolonged acquisitions and increase the proportion of analyzable datasets to nearly 100% in lymphoid and non-lymphoid tissues. Considering the straightforward implementation of our newly developed system, we anticipate that it will be applicable for other users interested in improving the quality of live imaging data acquisition.
authors: Albert Einstein Center for Fundamental Physics, Laboratory for High Energy Physics (LHEP), University of Bern, Sidlerstrasse 5, 3012 Bern, Switzerland. Electronic address: mykhailo.vladymyrov@lhep.unibe.ch.
authors: Mykhailo
authors: M
authors: Vladymyrov
authors: Theodor Kocher Institute, University of Bern, Freiestrasse 1, 3012 Bern, Switzerland.
authors: Jun
authors: J
authors: Abe
authors: Theodor Kocher Institute, University of Bern, Freiestrasse 1, 3012 Bern, Switzerland.
authors: Federica
authors: F
authors: Moalli
authors: Theodor Kocher Institute, University of Bern, Freiestrasse 1, 3012 Bern, Switzerland.
authors: Jens V
authors: JV
authors: Stein
authors: Albert Einstein Center for Fundamental Physics, Laboratory for High Energy Physics (LHEP), University of Bern, Sidlerstrasse 5, 3012 Bern, Switzerland.
authors: Akitaka
authors: A
authors: Ariga
mesh_terms: D019264:Adoptive Transfer; D059925:Anatomic Landmarks; D000818:Animals; D002465:Cell Movement; D003199:Computer Systems; D007091:Image Processing, Computer-Assisted; D000069416:Intravital Microscopy / Q000295:instrumentation*; D051379:Mice; D008810:Mice, Inbred C57BL; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D012867:Skin / Q000000981:diagnostic imaging*; D013601:T-Lymphocytes / Q000166:cytology*
keywords: GPU accelerated data processing; Intravital microscopy; Real-time image processing; Tissue drift