Associated Techniques
Circular Dichroism
LD is very similar to Circular Dichroism (CD), but with two important differences. (i) CD spectroscopy uses circularly polarized light whereas LD uses linearly polarized light. (ii) In CD experiments molecules are usually free in solution so they are randomly oriented. The observed spectrum is then a function only of the chiral or asymmetric nature of the molecules in the solution. With biomacromolecules CD is particularly useful for determining the secondary structure. By way of contrast, in LD experiments the molecules need to have a preferential orientation otherwise the LD=0. With biomacromolecules flow orientation is often used, other methods include stretched films, magnetic fields, and squeezed gels. Thus LD gives information such as alignment on a surface or the binding of a small molecule to a flow-oriented macromolecule, endowing it with different functionality from other spectroscopic techniques. The differences between LD and CD are complementary and can be a potent means for elucidating the structure of biological molecules when used in conjunction with one another, the combination of techniques revealing far more information can a single technique in isolation. For example CD tells us when a membrane peptide or protein folds whereas LD tells when it inserts into a membrane.
Fluorescence detected Linear Dichroism
Fluorescence-detected linear dichroism (FDLD) is a very useful technique to the experimentalist as it combines the advantages of UV LD whilst also offering the confocal detection of the fluorescence emission. FDLD has applications in microscopy, where can be used as a means of two-dimensional surface mapping through differential polarisation spectroscopy (DPS) where the anisotropy of the scanned object allows an image to be recorded. FDLD can also be used in conjunction with intercalating fluorescent dyes (which can also be monitored using UV LD). The intensity difference recorded between the two types of polarised light for the fluorescence reading is proportional to the UV LD signal, allowing the use of DPS to image surfaces
Read more about this topic: Linear Dichroism
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