Long-term time-lapse multimodal microscopy for tracking cell dynamics in live tissue
2011
High speed intravital microscopy has emerged as an essential tool for studying cellular dynamics in live tissue. A
limitation of this technique, however, is that the timescale that a sample can be continuously imaged is limited by
practical considerations to several hours. Long term observation of live tissue is of great interest for a variety of research
areas. We present methods for observing long term cellular dynamics in live tissue based on three-dimensional
registration of time-lapse intravital microscopy images.
For these experiments we utilized a custom multimodal microscope that allows simultaneous and co-registered
acquisition of optical coherence (OCM) and multiphoton (MPM) microscopy images. OCM allows the structure of a
sample to be visualized based on backscattered light while MPM excited fluorescence allows individual cells and cell
function to be visualized. The OCM images of tissue structure are used to register data sets taken at different time
points. The transformations of the OCM images are applied to MPM images to determine the migration of cell
populations. This method of image registration is applied to in vivo tracking of bone-marrow derived GFP-labeled stem
cells in mouse skin following bone marrow transplants from GFP donors into species-matched wildtype hosts. The use
of three-dimensional image registration of time-lapse microscopy images enables tracking these cells after local
cutaneous injury, and for investigating the role of skin stem cells in wound healing.
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