How do you remove the absorption or fluorescence of buffer from the total spectrum?
How do you generate the fluorescence spectrum of phycobiliproteins using Vernier Spectro Vis plus spectrometer? What is the light source you use for excitation?
Answers
Explanation:
Fluorescence spectroscopy is routinely used for studying structural changes in conjugated systems, aromatic molecules, and rigid, planar compounds due to alterations in temperature, pH, ionic strength, solvent, and ligands. A single fluorophore can generate thousands of detectable photons that can be repeatedly excited and detected, making fluorescence spectroscopy is a highly sensitive technique.
Fluorescence is a type of radiative emission that occurs when a molecule absorbs energy at a wavelength where it has a transition dipole moment. The excitation energy provided to the molecule at the ground state promotes photons to an excited singlet state, where they then decay to the lowest vibrational energy level of this excited singlet state. This energy further relaxes back to the ground state of the molecule, emitting photons in the process, as shown in Figure 1.
Fluorescence spectroscopy