Instantaneous, high-resolution 3D imaging using multifocus microscopy (MFM) and applications in functional neuronal imaging
Sara Abrahamsson
(Rockefeller University)
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Date: June 18, 2015
Description:
Multifocus microscopy (MFM) is an optical imaging technique that delivers instant 3D data at high spatial and temporal resolution. Diffractive Fourier optics are used to multiplex and refocus light, forming a 3D image consisting of an instantaneously formed focal stack of 2D images. MFM optics can be appended to a commercial microscope in a “black box” at the camera port, to capture 3D movies of quickly moving systems at the native resolution of the imaging system to which it is attached. MFM can also be combined with super-resolution methods to image beyond 200nm resolution. Systems can be tailored to fit different imaging volumes, covering for example seven or nine image planes to study mRNA diffusion inside a cell nucleus, or 25 focal planes or more to study cell division or neuronal activity in a developing embryo.
In the Bargmann lab at the Rockefeller University, MFM is applied in functional neuronal imaging in the nematode C. elegans – a millimeter-sized worm that with its compact but versatile nervous system of 302 neurons is a common model organism in neurobiology. The genetically expressed calcium-indicator dye GCaMP is used to visualize neuronal activity in the worm. Using MFM it is possible to image entire 3D clusters of neurons in living animals with single neuron resolution, and to perform unbiased imaging screenings of entire circuits to identify neuronal function during for example olfactory stimulation.
Further Information:
Sara came to UCSF in 2004 from the Royal Institute of Technology, Stockholm, Sweden to spend six months on an optical design project with Professor Mats Gustafsson. Mats tricked her into staying in the lab to do a Ph.D. and later to follow him to HHMI Janelia Research Campus. During this time, Sara developed two diffractive Fourier optics systems for live bio-microscopy in extended sample volumes: Extended Focus (EF)1 and multifocus microscopy (MFM)2,3. These techniques have since been applied to a wide variety of fast live-imaging projects in biological research.
Currently, Sara is a Leon Levy postdoctoral fellow in the laboratory of Cori Bargmann at the Rockefeller University, where she applies MFM to functional neuronal imaging in extended sample volumes. Sara also spends a lot of her time in the nanofabrication clean rooms at CNF, Cornell University, Ithaca, NY and NIST, MD, using multilevel, deep UV-lithography to fab the various diffractive Fourier optics devices she designs and builds in the labs of collaborators in the US and Europe. Sara spends her summers at the Marine Biological Laboratory in Woods Hole on Cape Cod, developing 3D polarization microscopy methods with the Oldenbourg lab.
Created: Tuesday, June 23rd, 2015