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Cellular Biology and Biochemistry > Fluorescence > Flashcards

Fluorescence Flashcards

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1
Q

What do macromolecules absorb?

A

Electromagnetic radiation

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2
Q

How many molecules can spectroscopy measure?

A

Multiple

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3
Q

What’s one way to measure reaction kinetics?

A

Coupling to a known chromophore

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4
Q

What is the Beer-Lambert relationship?

A

Absorbance(lambda) = epsilon(lambda) x concentration x path length

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5
Q

What wavelength do peptide bonds absorb?

A

214nm

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6
Q

What wavelength do aromatic amino acids absorb?

A

280nm

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7
Q

What is protein absorption affected by?

A

The microenvironment

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8
Q

What wavelength do nucleic acids absorb?

A

~260nm

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9
Q

What does absorbance do as DNA is heated?

A

Absorbance increases as temperature increases due to the change from double stranded to single stranded

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10
Q

What is Tm?

A

Melting temperature

The value at which 50% has denatured

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11
Q

Give a use of Tm

A

The ratio of A-T and G-C base pairs

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12
Q

What is the formula for energy absorbed?

A

Eabsorbed = heat x concentration/wavelength

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13
Q

What is the formula for energy emitted?

A

hc/lambda

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14
Q

How is energy lost when the excited electron moves back to the ground state?

A

As heat and as fluorescence

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15
Q

Describe the relationship between energy and wavelength

A

Lower energy = longer wavelength

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16
Q

What spectra do fluorescent molecules have?

A

Absorption and emission

17
Q

What is the Stokes shift?

A

The difference between the wavelengths of absorption and emission

18
Q

What is more sensitive? Fluorescence or absorbance

A

Fluorescence

19
Q

What is the set up of a basic fluorometer?

A

Light source
Monochromator
Sample
Sensor

20
Q

Name some uses of fluorescence

A 
Cellular localisation
Biosensor
Gene expression
Membrane potentials
Cell separation and counting
Ca2+ imaging
21
Q

What is GFP?

A

Green fluorescent protein

Emission at 509nm

Cellular Biology and Biochemistry (22 decks)

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