Fluorescence methods
in life sciences
Ctirad Hofr
lablifeB.org
1. What is the main topic of lectures?
2. Where can be knowledge used?
3. How will be course organizes?
Overview
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Why biophysical and
spectroscopy methods?
• We can quantify phenomena and compare
different biological systems
Why fluorescence?
• We can see things that would have been
hidden
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5 4
Literature
• Lakowicz J.R.: Principles of Fluorescence
Spectroscopy. Third Edition, Springer +
Business Media, New York, 2006.
Grafika z knihy Principles o Fluorescence byla pro účely této přednášky laskavě poskytnuta
profesorem J.R. Lakowitzem.
Poděkování
Fluorescence methods in life sciences
Practical applications of theoretical knowldege
Fluorescently labeled DNA Labeled protein separation
Biniding of DNA and proteinu
Anizotropie fluorescence
EmiseEmiseEmise
detektor
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−
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2II
II
r
ΙΙ
ΙΙpolarizátor
analyzátor
Where can you use obtained
knowledge?
• Practical fluorescence spectroscopy
• When you have small amount of your sample
• Everyday laboratory tasks (gel electrophoresis,
microscopy)
• Practicals C7235 !
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Multicolor Detection:
Image
Stain Target Color
DAPI Nucleii Blue
BODIPY® FL phallacidin F-actin Green
MitoTracker® Red CMXRos Mitochondria Orange 1/7
Flow Cytometry
Single cell detection
Cell sorting based on size and
fluorescence marker presence.
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Fluorescence at unexpected places
Is fluorescence intrinsic (natural) or extrinsic (synthetic)?
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Task = case studies
• Four tasks
• Points obtained for tasks contribute to exam
test score
• If you pass all tasks, you can apply for oral
exam
• 48 h for task solution
Practicals
23.-25.5.2017
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What does shine at the discoteque?
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Home work
• Narrating 0:40 do 3:00
Introduction to Fluorescence
https://www.thermofisher.com/cz/en/home/supp
ort/tutorials.html#vid1
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