11. labelfree FRET

Question 1

Absorption-emission

Answer 1

Question 2

What is the fluorescence lifetime ?

Answer 2

Question 3

What is the fluorescence lifetime ?

100 molecules

Answer 3

Question 4

What defines 𝜏௙௟௨௢ ?

Answer 4

Question 5

Why is the fluorescence lifetime interesting ?

Answer 5

Question 6

particular environment

Answer 6

Question 7

Application: mapping the complex of RNA polymerase

Answer 7

Question 8

How to measure tau?

Answer 8

Question 9

How to measure tau scheme?

Answer 9

Question 10

Example of a fluorescence decay

Answer 10

Question 11

(Frequency-domain lifetime measurements)

Answer 11

Question 12

What is FLIM

Answer 12

Question 13

Confocal fluorescence microscopy: principle

Answer 13

Question 14

Point excitation – point detection: how to image??

Answer 14

Question 15

Why is FLIM interesting ?

Answer 15

Question 16

Example: autofluorescence FLIM of stomach tissue

Answer 16

Question 17

Why is FLIM interesting ?

Answer 17

Question 18

Example: imaging pH in skin

Answer 18

Question 19

Why is FLIM interesting ?

Answer 19

Question 20

Example: differentiation state of stem cells

Answer 20

Question 21

Other examples: viscosity

Answer 21

Question 22

Other example: Imaging 9 different probes via FLIM

Answer 22

Question 23

everybody does FRET

Answer 23

Question 24

FRET principle 1

Answer 24

Question 25

FRET principle2

Answer 25

Question 26

What does *compatible mean?

Answer 26

Question 27

Take home message

Answer 27

Question 28

FRET Jablonski-wise

Answer 28

Question 29

What defines kT ?

Answer 29

Question 30

What can FRET be used for ?

Answer 30

Question 31

FRET is super-resolution

Answer 31

Question 32

Most common way

Answer 32

Question 33

Also popular: acceptor photobleaching FRET

Answer 33

Question 34

Also popular: acceptor photobleaching FRET

Answer 34

Question 35

Also popular: FRET-FLIM

Answer 35

Question 36

IN oligomerization via FLIM

Answer 36

Question 37

Advanced FRET: absolute distances

Answer 37

Question 38

Popular example: real-time PCR

Answer 38

Question 39

Example: estrogen biosensor

Answer 39

Question 40

Example: phosphatase sensor

Answer 40

Question 41

Example: Groel-Groes interaction

Answer 41

Question 42

Example: single-molecule FRET

Answer 42

Question 43

Why use a fluorescence microscope for measuring protein structures?

Answer 43

Question 44

Example: Multidrug resistance transporter LmrP

Answer 44

Question 45

Microdevices for screening membrane protein structure

Answer 45

Question 46

Microscopy setup @ BIOMED

Answer 46

Question 47

Example: Combined electrophysiology & smFRET

Answer 47

Question 48

Example: Measuring subcellular mechanical forces w/ FRET

Answer 48

Question 49

FRET measurements: anywhere!

Answer 49

Question 50

Lifetime/FLIM measurements @KULeuven

Answer 50

Question 51

Straightforward advanced lifetime analysis

Answer 51

Question 52

Why single virus imaging ?

Answer 52

Question 53

The HIV-1 lifecycle – early steps

Answer 53

Question 54

Challenges when studying HIV integration

Answer 54

Question 55

Measuring E for single molecules

Answer 55

Question 56

Labeling and imaging IN for single virus FRET

Answer 56

Question 57

LEDGINs block HIV replication by blocking IN-LEDGF interactions, but do they do more than that?

Answer 57

Question 58

At the nuclear membrane, intasome structure changes

Answer 58

Question 59

Transcription factor LEDGF/p75 alters IN stoichiometry

Answer 59

Question 60

Murine leukemia virus

Answer 60

Question 61

MLV needs mitosis to access the chromatin

Answer 61

Question 62

Counting cells with nuclear viruses

Answer 62

Question 63

Interactions with BET are needed

Answer 63

Question 64

Model

Answer 64