AbuArish_CPPS325_March07_2025_Lecture 1_Upload (1) Flashcards
What is the focus of Lecture 1?
Fluorescence imaging concepts
What is the topic of Lecture 2?
Lipid modulation of CFTR distribution under health conditions
What does Lecture 3 cover?
CFTR behaviours modulation by proteins
What is discussed in Lecture 4?
CFTR behaviours in cystic fibrosis
What is introduced in Lecture 5?
Introduction to Integrated stress response
What is the subject of Lecture 6?
Epithelial stress response under health & Cystic Fibrosis conditions
What is the definition of a fluorescent molecule?
EGFP
What is the resolution limit in fluorescence confocal microscopy?
Defined by the focal plane and point spread function
What is the wavelength range for fluorescence?
300–800 nm
What characterizes the fluorescence of a molecule?
Its emission spectrum
What is vibrational relaxation time?
10^-14 s
What is the typical lifetime of a photon in fluorescence?
1-100 ns
What are the criteria for a molecule to be a fluorophore?
- Efficiently absorbs a photon
- Efficiently emits a photon with longer wavelength
- Emitted photon is within the visible light spectrum
What are examples of organic fluorescent dyes?
- DAPI
- Rhodamine
- Alexa dyes
- Cyanine dyes
What is the source of the green fluorescent protein (GFP)?
Aequorea Victoria jellyfish
What are the excitation and emission wavelengths for EGFP?
Ex. 488 nm, Em. 509 nm
What are color-shifted genetic derivatives of EGFP?
- EBP (blue)
- ECFP (cyan)
- EYFP (yellow)
- mOrange (orange)
- mCherry (red)
- mRaspberry (far red)
Who developed fluorescence confocal microscopy?
Marvin Minsky, 1957
What is photobleaching?
Photo-induced alteration of a fluorophore into a non-fluorescent molecule
What can reduce photobleaching?
Decreasing the amount of oxygen in solution
What does STED stand for?
Stimulated Emission Depletion
What is a key advantage of STED over confocal microscopy?
Provides much higher spatial resolution (50 nm in x-y)
What is a disadvantage of STED microscopy?
Requires very high laser power leading to more photobleaching
How does STED improve spatial resolution compared to confocal microscopy?
By 5-fold
