Dear
Telamond;
Great!, - You did quite a couple, even if not all very well.
(Not only what
Arkcon already corrected.)
Let’s “walk” through your last Posting (# 3):
Using an angle of 0° would result in a Spectrum mixed from “Excitation” (in real an Absorption) with the resulting “
Emission Spectrum”. And how would you now like to separate both, to get the wanted “Emission Spectrum”? But you can be lucky, because the molecules will emission the resulting Photons in all directions. So you are able to measure the “pure” "
Emission Spectrum” without ‘any’ other influence of the “
Excitation Spectrum”, if you measure by 90°.
And what's the difference in an excitationspectrum and an emissionspectrum in a spectrofluorimeter?
The emissionspectrum gives their peak at a lower wavelength due to Stoke's shift.
Arkcon has given you already a correction to your answer, but it is not complete.
Your answer tells me, that you not really caught what “
Fluorescence” is, and that you can measure it in a “
Fluorescence Spectrometer” or a “
Fluorometer”, because both term mean the same.
With the “Excitation Light” you excite a Electron to a higher (mostly the next) “Energy level” and finally the Electron returns to the “
Groundstate” and in case of “Fluorescence” it emits a Photon, and if its energy falls into the visible range you can see it with your eye.
But that means you have at least to spend the electron enough energy to reach its excited state, and that means “
Excitation Wavelength” must be at least as short, or even shorter as the wavelength you can measure as the “
Emission Wavelength”, because as shorter the Wavelength as higher the Energy (absorbed or emitted) will be.
Yes!, - Dear
Redskytonight820; - that’s also partially very true, that very seldom (-if-) both wavelength are varied at the same time. But quite often Excitation is done by using a certain rang of Wavelengths below the lowest “Emission Wavelength” you would like to record, because a higher “yield” of Photons. The second part of your answer must be ‘headline-ed’: “
Only looks similar”, but that’s an additional theme.
But that’s not enough! As your “Groundstate”, as also your “
Excited State”, are overlaid by the corresponding “
Vibrational States” (a lot for each), you excite your electron into a lot of different “Vibrational States” of the “Excited State”, but the “Fluorescence” starts ‘all times’ from the lowest “Vibrational State”. So the electron has first to lose some of its “Excited Energy” to reach first the lowest “Vibrational State”; and that results in a so called “
Red Shift” of the “Emission Spectrum”, or in this case called “
Stokes Shift” (Direction to Read means less Energy).
(
Remember: The “Emission Spectrum” is not the
mirror of the “Excitation Spectrum”, as most people think/believe!)
Maybe for some visualisation you may visit: "
JABLONSKI Diagram”?
For the advantages of DAD’s /PAD’s over Photomultipliers you would have been able to find, under the Question “
How does UV/Vis Diode Array Spectrometer (PADs/DADs) work?”:
…………
E) CCD's Charge Coupled Device Detectors have, by their construction/electronics, only a few, and rarely required Advantages over PDA's/DAD's, but several Disadvantages!
F) Some other important "Adjectives":
• Single Beam (mostly). • No closed Sample Compartment required!
• Real Spectrum instead of Single Wavelength. • Predestinated for reliable Spectroscopy.
• Really fix Wavelength scale. • Only quasi equidistant Wavelength scale.
• Quasi fix Wavelength resolution. • Time stable Wavelength resolution!
• Exorbitant stable! (>15 Years/>30 Instr.!) • Predestinated for reliable Chemometrics.
• Predestinated for stable/secure Production. • Predestinated for validated Quality Control.
• etc.
G) etc.
Additional can be told, that you can, with a DAD, take a Spectrum of your Peaks for Identification. But not only, you can take a Spectrum at the start of your Peak, one at the maximum of your Peak, and one near the end of the peak. But for what? You can give the evidence that your analysed Peak is as good as you ever know, build from only “
One/Pure Compound”.
I think: Not 'nothing', because it costs you ‘nothing’, but gives you a lot.
The Spectra are also very important for
Validation/Identification of Method and Compound.
You can also ‘misuse’ DADs as a “
Multi-Single-Wavelength-Detector”, and so on, and so on.
A Disadvantage may be the shorter dynamic range in Absorbance (0.0 till ~1.8 AUs), but depends more on your calibration of your method.
But what a lot of people believe, that the “
Sensitivity” must to be lower is not true, and mostly caused by their wrong handling of the DAD.
Huuuch…..! I told you it will be a lot to write!
I hope it will be at least of some help to you.
Good Luck!
ARGOS
++