Course detail

Microscopic Imaging Technology

FEKT-FMZTAcad. year: 2010/2011

The course will be a detailed overview of the principle and practice of light microscopy. The emphasis of the course will be on the correct and appropriate use of the light microscope. Course covers optical microscope theory and also advanced optical and imaging techniques.

Language of instruction

Czech

Number of ECTS credits

6

Mode of study

Not applicable.

Guarantor

doc. Ing. Radim Kolář, Ph.D.

Department

Department of Biomedical Engineering (UBMI)

Learning outcomes of the course unit

Practical knowledge of physical principle, microscopy design concept and microscopic image processing in the field of light microscopy.

Prerequisites

The subject knowledge on the Bachelor´s degree level is requested.

Co-requisites

Not applicable.

Planned learning activities and teaching methods

Teaching methods depend on the type of course unit as specified in the article 7 of BUT Rules for Studies and Examinations.

Assesment methods and criteria linked to learning outcomes

Requirements for completion of a course are specified by a regulation issued by the lecturer responsible for the course and updated for every year.

Course curriculum

Fundamentals of wave and geometry optic.
Eye as an optical system.
Quantitative description of the optical system.
Basic microscopy design concept.
CCD and CMOS sensors.
Dark field microscopy.
Phase contrast microscopy.
Stereomicroscopy.
Nomarsky differential interference contrast microscopy.
Hoffman modulation contrast microscopy.
Fluorescence microscopy.
Laser scanning (confocal) microscopy.
2-photon and multi-photon microscopy.
Optical coherent microscopy.
Next advanced microscopic techniques - TIRFM, FRAP, FRET, STED.
Application of light microscopic principles in ophthalmology, dermatology, endoscopy.
Basic techniques in microscopic image processing.

Work placements

Not applicable.

Aims

Knowledge of the light microscopy and its medical and biologic applications. Practical knowledge about microscopic image acquisition and processing.

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

Limitations of controlled teaching and its procedures are specified by a regulation issued by the lecturer responsible for the course and updated for every year.

Recommended optional programme components

Not applicable.

Prerequisites and corequisites

Not applicable.

Basic literature

D. B. Murphy Fundamentals of light microscopy and electronic imaging, Wiley-Liss, 2001
P. Mouroulis Visual Instrumentation, McGraw-Hill, 1999

Recommended reading

Not applicable.

Classification of course in study plans

  • Programme BTBIO-F Master's

    branch F-BTB , 1. year of study, winter semester, compulsory

Type of course unit

 

Lecture

26 hours, optionally

Teacher / Lecturer

Mgr. Dušan Hemzal, Ph.D.

Syllabus

1. Wave and geometry optic - basic optical laws and phenomenon (interference, aberration, diffraction, polarization), optical components. Interaction of light with tissue - absorption, diffraction, attenuation, fluorescence, phosphorescence, autofluorescence.

2. Eye as an optical system, which participate on imaging process. Eye anatomy. Some rules connected to vision process (scotopic/photopic vision, Weber - Fencher law, Stiles - Crawford effect, darkness adaptation)

3. Description of the optical system. Quantitative evaluation of these systems (optical transfer function, modulation transfer function, Strehl ratio, wavefront aberration)

4. Basic microscopy design concept. Description and properties of particular components - holder, eyepiece, lens, condenser, light sources. Examples of microscopes.

5. Analog and digital microscopy. Light detection - CCD and CMOS sensors and their properties (signal-to-noise ratio, spatial resolution, temporal resolution). Videomicroscopy.

6. Upright and inverted microscopy - differences. Dark field microscopy - principle, design, applications. Phase contrast microscopy - physical and mathematic description, design, application.

7. Stereomicroscopy - principle, design, image processing. Nomarsky differential interference contrast (DIC) microscopy, Hoffman modulation contrast (HMC) microscope.

8. Fluorescence microscopy - description of fluorescence, fluorescence dyes, principles, microscope design.

9. Laser scanning microscopy, laser scanning confocal microscopy - principles, spatial resolution. Fluorescence scanning microscopy, 2-photon and mulit-photon microscopy.

10. Optical coherent microscopy and tomography - phenomenon of light interference for tomographic imaging. Systems working in temporal and frequency domain. Applications.

11. Next advances microscopic techniques - Total Internal Reflection Fluorescence Microscopy (TIRFM), Fluorescence Recovery After Photobleaching (FRAP), Fluorescence Resonance Energy Transfer (FRET), Stimulated Emission Depletion (STED), holographic microscopy.

12. Preparation of microscopic samples. Live cell imaging - heart cell contractility. Application of light microscopic principles in ophthalmology, dermatology, endoscopy.

13. Basic techniques in microscopic image processing - disparity maps in steremicroscopy, deconvolution, formation of focus image from different focus images sequence.

Laboratory exercise

26 hours, compulsory

Teacher / Lecturer

Ing. Jan Odstrčilík, Ph.D.

Syllabus

1. Introductory laboratory - introduction to laboratory equipments, introduction to image acquisition and analysis software NIS - Elements.
2. Image acquisition of blood smear by microscope Nikon Eclipse E200 and digital camera Pixelink A662. Image analysis of blood cells in NIS- Elements software.
3. Phase contrast- image acquisition of various scenes by microscope Nikon E80-i and digital camera DS Fi-1 using phase contrast - zoom 40x a 100x (with immersion oil). Image analysis in NIS- Elements software.
4. Stereomicroscopy - acquisition of large particles by stereomicroscope Nikon SMZ 1500 and digital camera DS Fi-1. Sejmutí scény s drobnými částicemi a vyhodnocení počtu těchto částic v programu NIS- Elements.
5. Image acquisition of heart muscle by microscope Nikon Eclipse E200 and digital camera Pixelink A662. Image analysis of blood cells in NIS- Elements software.
6. Measurement of the heart cell contractility - demonstration laboratory.

The other activities

13 hours, compulsory

Teacher / Lecturer

doc. Ing. Radim Kolář, Ph.D.

Syllabus

Individual works, consultation with lecturer, solution and elaboration, classroom project presentation/defense.