A Fluorescence Microscope Enables In Best Sample Preparation
Posted: Jul 12, 2017
A fluorescence microscope is a specific optical microscope and it uses fluorescence and phosphorescence in place of absorption and reflection. It is used to study the physical properties of organic and inorganic substances where the image of it is produced with the fluorescence. These microscopes can be simple like the epifluorescence microscope or the complex confocal microscope that is designed to use optical sectioning. This produces better resolution of fluorescent image. Most of these microscopes used in life science are of epifluorescence design.
The Specific Requirements
The excitation wavelength of the light is focused on the object through the lens and fluorescence emitted by it is focused on the detector. If you want to study an object under a Fluorescence Microscope you must make sure that the sample is itself fluorescent. For this you will have to create fluorescent sample and there are several methods of doing it. The primary technique followed is by labeling the sample with fluorescent stains. You can also use in case it is a biological sample an expression of fluorescent protein. Alternatively, you can also use the intrinsic fluorescence or auto-fluorescence of the sample.
The Biological Fluorescent Stains
This type of microscopy is a very powerful tool in life science. This allows specific and sensitive staining of the sample so that you can easily detect distribution of proteins, other molecules in the sample and much more. There are many such fluorescent stains designed for different biological molecules. These may be small intrinsically fluorescent molecules that bind with a molecule like the nucleic acid stains. However, there are other microscopes like the Inverted Microscope that are also used for sampling different objects in life science.
Different Fluorescent Molecules
There are different fluorescent molecules that are studied under a Fluorescence Microscope like the fluorochromes or fluorophores, fluorescein, DyLight 488 and Alexa Fluors. All these fluorescent molecules can be linked chemically with different molecules that help it to bind the target molecule of interest of the sample and result in proper imaging. There are also different techniques followed and one such technique is the Immunofluorescence. In this technique specifically high binding of antibody is used with the antigen. This helps in labeling specific proteins as well as other molecules inside the cell.
The Other Processes
In other processes followed the fluorophore is directly conjugated with the primary antibody of the sample. Alternatively, there may be a secondary antibody conjugated with fluorophore that binds specifically with the first used. Fluorescent protein is also used in modern genetics to modify DNA which enables easy carriage of the fluorescent protein reporter. This facilitates genetic modification of proteins. All this helps in easy tracking of the protein in the live cells of the targeted sample.
Annie Meiji is an owner with professional author of his business. He delivers fresh and important news on latest technology based Microscopes with their applications.