Size-exclusion chromatography (SEC) is a laboratory technique used to separate molecules based on their size. Recently, it has been used to separate liposomes – small vesicles composed of phospholipid membranes – into fractions based on their size and fluorescence. This article will explain why liposomes fluoresce during SEC.
What are Liposomes?
Liposomes are microscopic vesicles that are made up of a phospholipid bilayer membrane surrounding an internal aqueous compartment. They can be used as drug delivery vehicles since they can encapsulate hydrophilic and hydrophobic drugs, proteins, and other macromolecules. In the context of size-exclusion chromatography, liposomes may be used as molecular markers to monitor separation efficiency or to identify the purity of the sample being separated.
How Does Fluorescence Occur?
When light is shone upon liposome samples during size-exclusion chromatography, the liposomes fluoresce because of the presence of fluorescent molecules within them such as alkyl esters and polycyclic aromatic hydrocarbons. These fluorescent molecules absorb energy from incoming light and re-emit it at longer wavelengths, resulting in a visible fluorescence or glow. By measuring this fluorescence intensity at different points along the separation column, scientists can track how efficiently the liposomes have been separated into fractions by size-exclusion chromatography.
The Benefits of Fluorescence During SEC
Fluorescent molecules provide several benefits when performing size-exclusion chromatography on liposome samples. Firstly, they allow scientists to more easily distinguish between fractions with similar sizes but different compositions. Secondly, they enable scientists to monitor changes in lipid composition during sample preparation or storage prior to analysis which could affect the accuracy of results obtained from SEC experiments. Finally, by tracking changes in fluorescence intensity over time, researchers can identify potential problems with their separation columns before they become too severe which could interfere with accurate analysis results.
Conclusion:
Liposomes are tiny vesicles composed of phospholipid membranes that are often used for drug delivery purposes due to their ability to encapsulate both hydrophobic and hydrophilic substances within them. During size exclusion chromatography (SEC), these lipids will fluoresce due to fluorescent molecules present within them such as alkyl esters and polycyclic aromatic hydrocarbons which absorb energy from incoming light and re-emit it at longer wavelengths resulting in visible fluorescence or glow. This fluorescence provides several benefits including allowing scientists to more easily distinguish between fractions with similar sizes but different compositions as well as monitoring changes in lipid composition during sample preparation or storage prior to analysis which could affect accuracy results obtained from SEC experiments. With this knowledge about why liposomes fluoresce during SEC experiments, researchers can better understand how best to use this technique for accurate data collection on lipids samples for drug delivery purposes or other research needs involving these types of particles.
