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Depth-Resolved Optical Nanoscopy

Objective

A fluorescence microscope for the noninvasive imaging of the structure and dynamics in thick 3D systems with 3D sub-diffraction-limited resolution and at extended penetration depths would have a broad spectrum of applications in both the physical and biological sciences. Conventional fluorescence microscopy techniques, such as confocal and two-photon microscopy, lack the spatial resolution required for measurements at the nanoscale, whereas state-of-the-art 3D super-resolution optical microscopy methods offer limited penetration depth (<5 microns). In the proposed project, I aim to develop a new microscope that combines molecular photoswitches, fluorescence self-interference and light-sheet microscopy to optically section a thin layer deep in thick semitransparent samples with 3D sub-diffraction-limited resolution. I will then test it for potential physical and biological applications of single molecule detection. The first specific aim is to develop a depth-resolved self-interference photoswitching nanoscope employing novel high-throughput and sub-diffraction-limit fluorescence interferometry. The second specific aim is to map with 3D nanometer resolution the motion of photoswitchable fluorescent probes deep in soft materials. This study will open up new possibilities for precise measurements of the heterogeneity and mechanical properties of the nanoenvironments of soft matter at extended depths and may ultimately assist in developing superior biomaterials and nanomedical therapeutics. The third specific aim is to in situ image with 3D nanometer resolution the muscle thick filament system in fixed Caenorhabditis elegans nematodes at both the ventral and dorsal ends (separated by ~80 microns). This demonstration, if successful, will provide unique 3D super-resolution in situ image data and may assist in developing imaging protocols for in situ nanoscopy of structural changes of the C. elegans body-wall muscle system due to sarcomeric muscle diseases.

Field of science

  • /natural sciences/biological sciences
  • /natural sciences/physical sciences/optics/microscopy/super resolution microscopy

Call for proposal

FP7-PEOPLE-2011-CIG
See other projects for this call

Funding Scheme

MC-CIG - Support for training and career development of researcher (CIG)

Coordinator

BEN-GURION UNIVERSITY OF THE NEGEV
Address
.
84105 Beer Sheva
Israel
Activity type
Higher or Secondary Education Establishments
EU contribution
€ 100 000
Administrative Contact
Daphna Tripto (Ms.)