Course detail

Light Microscopy

FSI-9MIKAcad. year: 2020/2021

The course explains principles of light microscopy in terms of geometrical and wave optics. Imaging of extended objects is analysed in details using the formalism of pupil and transfer functions. Fundamentals of advanced microscopis methods are explained as well as the directions of their development (confocal microscopy, fluorescence microscopy, interference microscopy, nonlinear phenomena, Raman scattering, stimulated light emission).

Learning outcomes of the course unit

Knowledge of fundamentals of light microscopy and competence to apply them in practice.


Wave and Fourier optics, geometrical optics.


Not applicable.

Recommended optional programme components

Not applicable.

Recommended or required reading
L. C. Martin: The Theory of the Microscope.. Blackie, London 1966
D. B. Murphy: Fundamentals of light microscopy and electronic imaging. Wiley-Liss, Hoboken 2001. (EN)
M. Born, E. Wolf: Principles of Optics. 7th (expanded) ed.. Cambridge University Press, Cambridge 1999
Born, M., Wolf, E.: Principles of Optics. 7th (expanded) ed.. Cambridge University Press, Cambridge 1999 (EN)
J. W. Goodman: Introduction to Fourier Optics. 2nd ed.. McGraw-Hill, New York 1996
A. R. Hibbs: Confocal Microscopy for Biologists. Springer, 2004. (EN)
E. Keprt: Teorie optických přístrojů 2, Teorie a konstrukce mikroskopu, SPN, Praha 1966.
T. Wilson: Confocal Microscopy. Academic Press, London 1990 (EN)

Planned learning activities and teaching methods

The course is taught through lectures explaining the basic principles and theory of the discipline. The lecture is completed by advanced optical demonstrations and experiments with light microscopes.

Assesment methods and criteria linked to learning outcomes

Examination is oral. Detailed knowledge of explained matter is proved.

Language of instruction

Czech, English

Work placements

Not applicable.


Understanding of principles of light microscopy and microscopic imaging, knowledge of conventional and advanced microscopic techniques.

Specification of controlled education, way of implementation and compensation for absences

Presence at lectures is not obligatory, but recommended.

Classification of course in study plans

  • Programme D-MAT-P Doctoral, 1. year of study, summer semester, 0 credits, recommended

  • Programme D4F-P Doctoral

    branch D-FMI , 1. year of study, summer semester, 0 credits, recommended

Type of course unit



20 hours, optionally

Teacher / Lecturer


1. Light as the electromagnetic wave, coherence of light, Hopkins' formula.
2. Fundamentals of geometric optics, optical abberations, elementary parts of the microscope.
3. Historical overview.
4. Wave description of imaging, the Abbe theory, the impulse response.
5. Coherent and optical transfer function.
6. Basic techniques of conventional microscopy, bright and dark field.
7. Phase contrast.
8. Polarized light.
9. Confocal microscopy.
10. Interference microscopy.
11. Fluorescence microscopy, modern techniques of microscopy.