MultiFocus Polarization Microscope (MF-PolScope) for 3D polarization imaging of up to 25 focal planes simultaneously

Sara Abrahamsson1, Molly McQuilken2, Shalin B. Mehta3, Amitabh Verma3, Johannes Larsch1, Rob Ilic1, Rainer Heintzmann4,
Cornelia I. Bargmann1, Amy S. Gladfelter2, and Rudolf Oldenbourg3

1The Rockefeller University, New York, New York, USA
2Department of Biological Sciences, Dartmouth College, Hanover, NH 03755
3Cellular Dynamics Program, Marine Biological Laboratory, Woods Hole, MA 02543
4 Friedrich-Schiller-Universität Jena, Helmholtzweg 4, 07743 Jena, Germany

We have developed an imaging system for 3D time-lapse polarization microscopy of living biological samples. Polarization imaging reveals the position, alignment and orientation of submicroscopic features in label-free as well as fluorescently labeled specimens. Optical anisotropies are calculated from a series of images where the sample is illuminated by light of different polarization states. Due to the number of images necessary to collect both multiple polarization states and multiple focal planes, 3D polarization imaging is most often prohibitively slow. Our MF-PolScope system employs multifocus optics to form an instantaneous 3D image of up to 25 simultaneous focal-planes. We describe this optical system and show examples of 3D multi-focus polarization imaging of biological samples, including a protein assembly study in budding yeast cells.

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