Primary Faculty

Mats Gustoffson

New Forms of Biological Light Microscopy

Research Description

Structured illumination microscopy

The light microscope is a ubiquitous tool in biology, but its spatial resolution is limited by diffraction. Several of our projects use a simple trick to circumvent the diffraction limit: structured illumination microscopy (SIM). SIM works by illuminating the specimen by a pattern of illumination light, which causes otherwise unobservable high-resolution information to be come observable in the form of low-resolution moiré fringes.

Nonlinear structured illumination microscopy

Even much higher resolution – in theory, unlimited resolution – is possible by exploiting nonlinear phenomena to generate harmonics of the physical illumination intensity pattern. We have demonstrated this concept in a simple form, where the nonlinearity stemmed from saturation of the excited state. We are now developing other forms, more suitable for biology, in which the nonlinearity is instead generated through photo-switching of fluorescent molecules.

Rapid live structured illumination microscopy

The ability to view living specimens is one of the great strengths of the light microscope. To allow us to apply the resolving power of structured illumination to living specimens, we are developing ways to increase the acquisition speed greatly, to the point where only negligible sample motion can take place during each data set.

Imaging rapid 3D processes

Even at conventional resolution, it is sometimes impossible to acquire data fast enough to follow a dynamic process in three dimensions. We are working on different tricks to allow a microscope to image a 3D volume instantaneously, without going through the normal process of acquiring a series of images at different focus.

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Current Projects

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Selected Publications

A New Approach to Extended Focus for High-Speed, High-Resolution Biological Microscopy. S. Abrarahmson, S. Usawa, and M.G.L. Gustafsson: Proc. SPIE 6090, 60600N-1-8 (2006). Download PDF

Nonlinear Structured-Illumination Microscopy: Wide-Field Fluorescence Imaging with Theoretically Unlimited Resolution. M.G.L. Gustafsson: Proc. Nat. Acad. Sci. USA. 102 13081–6 (2005). Download PDF

Phase-Retrieved Pupil Functions in Wide-Field Fluorescence Microscopy. B. Hanser, M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: J. Microsc., 216, 32–48 (2004). Download PDF

Phase Retrieval for High Numerical Aperture Optical Systems. B. Hanser, M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: Opt. Lett. 28, 801–3 (2003). Download PDF

Application of Phase Retrieved Pupil Functions in Wide-Field Microscopy. B. Hanser, M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: Proc. SPIE 4621, 40–6 (2002). Download PDF

Computational Adaptive Optics for Live Three-dimensional Biological Imaging. Z. Kam, B. Hanser, M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: Proc. Nat. Acad. Sci. USA 98, 3790–5 (2001). Download PDF

Phase Retrieval of Wide-Field Microscopy Point Spread Functions. B. Hanser, M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: Proc. SPIE 4261, 60–8 (2001).

Surpassing the Lateral Resolution Limit by a Factor of Two using Structured Illumination Microscopy. M.G.L. Gustafsson: J. Microsc. 198, 82–7 (2000). Download PDF - Download Cover Page

Doubling the Lateral Resolution of Wide-field Fluorescence Microscopy using Structured Illumination. M.G.L. Gustafsson, D. A. Agard, and J. W. Sedat: Proc. SPIE 3919, 141–50 (2000). Download PDF

Extended Resolution Fluorescence Microscopy. M.G.L. Gustafsson: Curr. Opin. Struct. Biol. 9, 627–34 (1999). Download PDF

I5M: 3D Widefield Light Microscopy with Better Than 100 nm Axial Resolution. M.G.L. Gustafsson, D.A. Agard, and J.W. Sedat: J. Microsc. 195, 10–16 (1999). Download PDF

3D Widefield Microscopy Using Two Objective Lenses: Experimental Verification of Improved Axial Resolution. M.G.L. Gustafsson, D.A. Agard, and J.W. Sedat: Proc. SPIE 2655, 62-6 (1996).

Sevenfold Improvement of Axial Resolution in 3D Widefield Microscopy Using Two Objective Lenses. M.G.L. Gustafsson, D.A. Agard, and J.W. Sedat: Proc. SPIE 2412, 147-56 (1995). Download PDF

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Contact Information

Email mats@msg.ucsf.edu
Phone (415) 514-4385

Mailing Address
Dept. of Physiology and Program in Bioengineering
University of California, San Francisco
Room BH-303, UCSF Box 2532
1700 4th Street
San Francisco, CA 94143-2532

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