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Describe your method for determining optimum λex and λem values in flourescence spectrometry.

Describe your method for determining optimum λex and λem values in flourescence spectrometry.

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With the help of UV-Vis spectroscopy, first we need to measure the wavelength at which the molecule has maximum absorption (ie. λmax). Now in Fluorescence Spectroscopy, this λmax is treated as excitation wavelength (λex)and therefore we need to set the instrument with λex to excite the molecule under examination. Due to this, molecule will absorb the λex and gets excited and while coming back to the ground sate it release the energy in the form of light which is nothing but maximum emission wavelength (λem) and can be measured on the instrument.

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