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The best cuvettes need to be very clear and have no impurities that might affect the spectroscopic reading. Defects on the cuvette such as scratches, can scatter light and hence should be avoided. Some cuvettes are clear only on two sides, and can be used in the UV-Visible spectrophotometer, but cannot be used for fluorescence spectroscopy measurements. For Group 12-16 semiconductor nanoparticles prepared in organic solvents, the quartz cuvette is chosen.

In the sample cell the quantum dots are dispersed in a solvent, whereas in the reference cell the pure solvent is taken. It is important that the sample be very dilute (maximum first exciton absorbance should not exceed 1 au) and the solvent is not UV-visible active. For these measurements, it is required that the solvent does not have characteristic absorption or emission in the region of interest. Solution phase experiments are preferred, though it is possible to measure the spectra in the solid state also using thin films, powders, etc. The instrumentation for solid state UV-visible absorption spectroscopy is slightly different from the solution phase experiments and is beyond the scope of discussion.

Detector

Detector converts the light into a current signal that is read by a computer. Higher the current signal, greater is the intensity of the light. The computer then calculates the absorbance using the in [link] , where A denotes absorbance, I is sample cell intensity and I o is the reference cell intensity.

The following cases are possible:

  • Where I<I 0 and A<0. This usually occurs when the solvent absorbs in the wavelength range. Preferably the solvent should be changed, to get an accurate reading for actual reference cell intensity.
  • Where I = I and A= 0. This occurs when pure solvent is put in both reference and sample cells. This test should always be done before testing the sample, to check for the cleanliness of the cuvettes.
  • When A = 1. This occurs when 90% or the light at a particular wavelength has been absorbed, which means that only 10% is seen at the detector. So I 0 /I becomes 100/10 = 10. Log 10 of 10 is 1.
  • When A>1. This occurs in extreme case where more than 90% of the light is absorbed.

Output

The output is the form of a plot of absorbance against wavelength, e.g., [link] .

Representative UV-visble absorption spectrum for CdSe tetrapods.

Beer-lambert law

In order to make comparisons between different samples, it is important that all the factors affecting absorbance should be constant except the sample itself.

Effect of concentration on absorbance

The extent of absorption depends on the number of absorbing nanoparticles or in other words the concentration of the sample. If it is a reasonably concentrated solution, it will have a high absorbance since there are lots of nanoparticles to interact with the light. Similarly in an extremely dilute solution, the absorbance is very low. In order to compare two solutions, it is important that we should make some allowance for the concentration.

Effect of container shape

Even if we had the same concentration of solutions, if we compare two solutions – one in a rectagular shaped container (e.g., [link] ) so that light travelled 1 cm through it and the other in which the light travelled 100 cm through it, the absorbance would be different. This is because if the length the light travelled is greater, it means that the light interacted with more number of nanocrystals, and thus has a higher absorbance. Again, in order to compare two solutions, it is important that we should make some allowance for the concentration.

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Source:  OpenStax, Nanomaterials and nanotechnology. OpenStax CNX. May 07, 2014 Download for free at http://legacy.cnx.org/content/col10700/1.13
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