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CZECH TECHNICAL UNIVERSITY IN PRAGUE
STUDY PLANS
2023/2024
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Methods of Imaging Tissue Cultures and Biological Structures

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Code Completion Credits Range Language
F7DIMZKS ZK 20P+8C Czech
Garant předmětu:
Roman Matějka
Lecturer:
Roman Matějka
Tutor:
Daniel Hadraba, Roman Matějka
Supervisor:
Department of Biomedical Technology
Synopsis:

Methods of cell culture and tissue imaging, problems of imaging and contrast enhancement, light and epi-light microscopy, fluorescent labeling, primary and secondary antibodies, conjugates, permeabilization and blocking of samples against non-specific components. Methods of fluorescence and confocal microscopy, imaging of complex structures.

Requirements:

As a standard, teaching takes place in contact form and the course ends with an oral exam, which is preceded by written preparation. If the number of students is less than 5, teaching can take place in the form of guided self-study with regular consultations. Furthermore, it is required to prepare a written study by the student on a given topic in the field. The condition for admission to the exam is the completion of two laboratory exercises (evidenced by a protocol signed by the student, the head of the exercise and the guarantor of the course). These protocols will be archived.

Syllabus of lectures:

Syllabus of lectures:

1. Light, epi-light microscopy,

2. Problematic tissue imaging, sample preparation

3. Fluorescence and phosphorescence

4. Fluorescence labeling of antibodies, blocking and permeabilization

5. Primary and secondary antibodies, conjugates

6. Imaging of intra and extracellular proteins

7. Confocal microscopy - LASER scanning

8. Confocal microscopy - CARV spinning disk

9. Multifot emission, second harmonic generation

10. SHG methods and use for protein imaging

Syllabus of tutorials:

Syllabus of exercises (block form of teaching after lessons):

1. Imaging of native biological structures, phase contrast, sample fixation, membrane permeabilization, blocking of non-specific bonds, preparation of primary and secondary antibodies, staining of samples, staining of nuclei

2. Preparation of widefield fluorescence microscope, setting of excitation, exposure parameters, scanning, preparation of confocal microscope, setting of excitation parameters, pinhole, scanning of Z-stack and multitile sample

Study Objective:

Methods of cell culture and tissue imaging, problems of imaging and contrast enhancement, light and epi-light microscopy, fluorescent labeling, primary and secondary antibodies, conjugates, permeabilization and blocking of samples against non-specific components. Methods of fluorescence and confocal microscopy, imaging of complex structures.

Study materials:

[1] Amelinckx,S., D. van Dyck, J. van Landuyt, G. van Tendeloo. Handbook of Microscopy: Applications in Materials Science, Solid-State Physics and Chemistry Applications.VCH Verlagsgesellschaft mbH, 1997.

[2] Lanza, R., Langer, R., Vacanti, J., Principles of Tissue Engineering, ed. 3rd Edition , Elsevier Academic Press, 2007, ISBN 978-0123706157

Note:
Time-table for winter semester 2023/2024:
Time-table is not available yet
Time-table for summer semester 2023/2024:
Time-table is not available yet
The course is a part of the following study plans:
Data valid to 2024-04-21
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