GFP

Question 1

What defines a fluorescent molecule?

Answer 1

One that can absorb light of a short wavelength and emit at a longer wavelength.

Question 2

What is a fluorophore?

Answer 2

A conjugated of aromatic structure that is the functional group in a fluorescent molecule

Question 3

What are the applications of fluorescence?

Answer 3

Real time PCR, fluorescent microscopy, flow cytometry, immunofluorescence using antibodies.

Question 4

What are the pros and cons of immunofluorescence.

Answer 4

Sensitive, enhances a weak signal, multicolour, flexible.

Samples has to be fixed and sample is often permeabilised for entry.

Question 5

What were the advantages of fluorescent proteins?

Answer 5

Increases speed of screening gene expression, no need to process if GFP is expressed, visualise using light microscope or UV lamp, allows protein movement and gene activity to be studied.

Question 6

What is FRET?

Answer 6

Fluorescence resonance energy transfer, uses a donor and acceptor chromophore measured the distance of energy transfer. A molecular ruler that can measure 2-8 nm.

Question 7

What colour does GFP absorb?

Answer 7

Blue, emits green