AbuArish_CPPS325_March07_2025_Lecture 1_Upload (1)

Question 1

What is the focus of Lecture 1?

Answer 1

Fluorescence imaging concepts

Question 2

What is the topic of Lecture 2?

Answer 2

Lipid modulation of CFTR distribution under health conditions

Question 3

What does Lecture 3 cover?

Answer 3

CFTR behaviours modulation by proteins

Question 4

What is discussed in Lecture 4?

Answer 4

CFTR behaviours in cystic fibrosis

Question 5

What is introduced in Lecture 5?

Answer 5

Introduction to Integrated stress response

Question 6

What is the subject of Lecture 6?

Answer 6

Epithelial stress response under health & Cystic Fibrosis conditions

Question 7

What is the definition of a fluorescent molecule?

Answer 7

EGFP

Question 8

What is the resolution limit in fluorescence confocal microscopy?

Answer 8

Defined by the focal plane and point spread function

Question 9

What is the wavelength range for fluorescence?

Answer 9

300–800 nm

Question 10

What characterizes the fluorescence of a molecule?

Answer 10

Its emission spectrum

Question 11

What is vibrational relaxation time?

Answer 11

10^-14 s

Question 12

What is the typical lifetime of a photon in fluorescence?

Answer 12

1-100 ns

Question 13

What are the criteria for a molecule to be a fluorophore?

Answer 13

• Efficiently absorbs a photon
• Efficiently emits a photon with longer wavelength
• Emitted photon is within the visible light spectrum

Question 14

What are examples of organic fluorescent dyes?

Answer 14

• DAPI
• Rhodamine
• Alexa dyes
• Cyanine dyes

Question 15

What is the source of the green fluorescent protein (GFP)?

Answer 15

Aequorea Victoria jellyfish

Question 16

What are the excitation and emission wavelengths for EGFP?

Answer 16

Ex. 488 nm, Em. 509 nm

Question 17

What are color-shifted genetic derivatives of EGFP?

Answer 17

• EBP (blue)
• ECFP (cyan)
• EYFP (yellow)
• mOrange (orange)
• mCherry (red)
• mRaspberry (far red)

Question 18

Who developed fluorescence confocal microscopy?

Answer 18

Marvin Minsky, 1957

Question 19

What is photobleaching?

Answer 19

Photo-induced alteration of a fluorophore into a non-fluorescent molecule

Question 20

What can reduce photobleaching?

Answer 20

Decreasing the amount of oxygen in solution

Question 21

What does STED stand for?

Answer 21

Stimulated Emission Depletion

Question 22

What is a key advantage of STED over confocal microscopy?

Answer 22

Provides much higher spatial resolution (50 nm in x-y)

Question 23

What is a disadvantage of STED microscopy?

Answer 23

Requires very high laser power leading to more photobleaching

Question 24

How does STED improve spatial resolution compared to confocal microscopy?

Answer 24

By 5-fold

Question 25

What does fluorescence super-resolution imaging allow for?

Answer 25

Spatial distinction of molecules separated by 50 nm

Question 26

What is the focal volume in fluorescence confocal microscopy?

Answer 26

250 nm

Question 27

What defines the resolution in fluorescence microscopy?

Answer 27

Point spread function (PSF)