Single-Shot Optical Sectioning Using Polarised Illumination Coded Structured Illumination Microscopy (picoSIM)

Student thesis: Doctoral ThesisDoctor of Philosophy


The conventional epi-uorescent wide-eld microscope features a uniform illumination of an extended sample region. A problem arises with this setup since light from outof-focus uorophores is also detected. The result is poor quality in the nal image, as out-of-focus structures appear blurred; furthermore, their emission light contributes to the background and leads to a reduction in image contrast.Removing out-of-focus light yields an optically sectioned image: a thin slice of a thick sample that only contains in-focus information. Taking a stack of such sectioned images allows for a three-dimensional (3D) view of the specimen. Structured illumination microscopy for sectioning (sSIM) is a method to obtain optically sectioned data, similar to that obtained from the widely used confocal microscope. However, sSIM suffers from a limited acquisition rate, as at least three individual raw images are needed to reconstruct one sectioned slice.
The technique of polarised illumination coded structured illumination microscopy
(picoSIM) combines optical sectioning with high temporal resolution. In picoSIM the
individual light patterns needed to acquire the raw sSIM images are encoded in the polarisation of the illumination light. This enables the simultaneous acquisition of the data needed for the sSIM reconstruction, allowing optical sectioning with high acquisition rates. This thesis describes the theory of picoSIM and presents experimental results.
Date of AwardNov 2013
Original languageEnglish
Awarding Institution
  • King's College London
SupervisorRainer Heintzmann (Supervisor) & Elisabeth Ehler (Supervisor)

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