M.Sc Thesis

M.Sc StudentShterman Doron
SubjectMultiple Moire' Structured Illumination Patterns for Spatial
Resolution Enhancement in Florescence Microscopy
DepartmentDepartment of Electrical and Computer Engineering
Supervisor PROF. Guy Bartal
Full Thesis textFull thesis text - English Version


Biological research nowadays is heavily dependent on the capabilities of modern microscopy, especially in terms of spatial and temporal resolution. Spatial resolution in conventional light microscopy is however limited by the well-known diffraction limit, as formulated by Ernst Abbe in 1873. Under this limitation standard optical microscopy can provide spatial separation of up to roughly 200 nm, which pose a significant limitation on modern Biological research.

Several ground breaking super-resolution techniques have evolved in the past decades based on fluorescence microscopy. Methods like stimulated emission depletion (STED) and photo-activated localization microscopy (PALM), can achieve sub 50 nm resolution yet involve scanning the sample thus require long acquisition times and \ or rely on high and damaging intensities, making them inadequate for dynamic in vitro studies. structured illumination microscopy (SIM), on the other hand, is a wide-field super-resolution technique with superior temporal capabilities, however conventionally has a resolution limit of roughly 100 nm, making it inferior to other super-resolution techniques.

In this dissertation we explore and demonstrate novel super-resolution microscopy concept, based on combination of structured illumination and total internal reflection techniques. We propose utilizing multi moiré illumination patterns and high index material in order to achieve sub 50 nm imaging while preserving SIM temporal resolution capability as well as its compatibility with living samples.

Following theoretical study, supported by an elaborated 2D simulation of the proposed optical scheme, we have successfully implemented and tested the multi moiré SIM, including a suitable reconstruction algorithm, proving its competence to produce up to 4-fold resolution enhancement beyond the diffraction limit.