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BIOC1007 CH > Fluorescence Spectroscopy > Flashcards

Fluorescence Spectroscopy Flashcards

1
Q

What is the difference between Fluorescence and phosphorescence?

A

A photon is emmited immediately in fluorescence, whereas in phosphorescence it is emitted after a while

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2
Q

Photons re-emitted after light absorption have a shorter or longer wavelength?

A

Longer wavelength, because some energy is lost when raising the energy level of the electron

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3
Q

Which amino acid residue if fluorescent?

A

Tryptophan/Trp/W

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4
Q

Why is fluorescence spectroscopy so sensitive?

A

Low background interference, so easier to detect very small amounts of photons

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5
Q

What light source is used in fluorescence spectroscopy?

A

A Xenon lamp

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6
Q

What is the name for a small molecule which can add fluorescence to a previously in-fluorescent molecule?

A

A fluorophore

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7
Q

What chemical in tonic water makes it fluorescent?

A

Quinine

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8
Q

What common effervescent beverage is fluorescent?

A

Tonic water

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9
Q

To what small molecule would Ethidium Bromide be used as a dye?

A

DNA

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10
Q

DNA is dyed by what molecule?

A

Ethidium Bromide

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11
Q

What molecules are fluorescein attached to, to cause fluorescence?

A

Proteins

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12
Q

What is attached to proteins to make them more fluorescent (does not include tryptophan)

A

Fluorescein

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13
Q

Compared to the incident light (I0), what direction do the IE (emission) and IA (absorption) light paths travel in?

A

IE (emission) is perpendicular to the incident light, IA (absorption) follows same direction as I0

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14
Q

What is the Stokes Shift?

A

The difference in wavelength between the maxima of IA and the maxima of IE

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15
Q

What is the name for the difference in maxima between IE and IA?

A

Stokes Shift

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16
Q

What relationship is there between the degree of fluorescence and concentration?

A

Directly proportional

17
Q

What does the degree of fluorescence depend on other than concentration of fluorescent molecule?

A

Environment around the fluorescent species, proximity to other species, this allows changes to be monitored

18
Q

What are the main uses of fluorescence spectroscopy?

A

DNA detection on gels, Labelling proteins, Protein dynamics, enzyme reaction rates

19
Q

What organism was GFP isolated from?

A

Jellyfish

20
Q

How is GFP bonded to a protein?

A

Covalently

21
Q

Roughly what shape is GFP

A

Similar to a beta-barrel

22
Q

What spectroscopies use: long, medium and short waves

A

long: NMR, medium EPR or UV/Vis, short X-ray absorption

BIOC1007 CH (7 decks)

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