Presentation Topics (and example questions) for Part 2: Optical coherence
tomography, Optical projection tomography, Confocal imaging, Super-
resolution techniques.
1. Describe the working principle of a time domain Optical Coherence Tomography (TD-OCT)
system. Compare this to a spectral domain OCT system and discuss the advantages and
disadvantages of each system. List possible applications of OCT.
Example questions:
▫ What are suitable samples for an OCT system, what limits its applicability?
▫ Which noise sources are present in typical OCT systems and what mitigation options
exist?
▫ Discuss factors limiting the depth-resolution of an OCT system; contrast with factors
that limit lateral resolution.
2. Describe how Optical Projection Tomography (OPT) imaging works, and 3D image
reconstruction via the Fourier Slice Theorem. Discuss it’s general advantages and
limitations, using a particular system (commercial or from the literature) as an example.
Example questions:
▫ Compare/contrast the OPT with confocal microscopy (with a focus on the capabilities
of OPT, as distinct from 1.a).
▫ What limits the resolution of the OPT system you chose to use as an example? How
could the design be improved to improve resolution?
▫ Justify whether or not it is possible to perform OPT of a continually changing sample.
3. Describe the working principle of a confocal microscope, in comparison with the widefield
fluorescence microscope. Discuss the definition of resolution in the context of confocal
microscopy and how the pinhole can influence the resolution.
Example questions:
▫ How do we decide the pinhole aperture for our experiment? What is the
recommended setting? Why?
▫ What techniques have been developed in widefield microscopy to improve resolution
beyond the diffraction limit?
4. “In confocal microscopy, there is a certain trade-off between spatial resolution, speed,
contrast, and depth penetration”. Outline the different aspects of this topic, clarifying in which
terms they are interconnected and how they can be controlled/varied experimentally.
Discuss one or two examples of hardware advance that improved the imaging performance.
Example questions:
▫ In biological applications, the sample viability (integrity) has great relevance. How
does this requirement influence the choice of the imaging technique and/or of the
imaging parameters?
▫ Can you guess how Optical Coherence Tomography generally compares with
Confocal Laser Scanning Microscopy in relation to the properties mentioned above?
