Fluorescence Microscopy is the technique of using excitation light to illuminate fluorescence molecules which are emitted at a different band of light in order to be imaged and analyzed. The emitted light is almost simultaneous with the excitation illumination.
The British scientist Sir George G. Stokes first described fluorescence in 1852 when he observed that the mineral fluorspar emitted red light when it was illuminated by ultraviolet excitation. He noted that the excitation light was always a shorter wavelength than the longer emitted fluorescence light. Further investigations in the 19th century led to the discovery of many specimens that fluoresce when irradiated. It wasn’t until the 1930’s when it was discovered that chemicals made up of fluorochromes can be used to dye biological tissues which led to the development of the fluorescence microscope.
Fluorescence microscopy has become an invaluable tool in the research of biomedical sciences as well as other chemical and material sciences based research. The advancement of advanced combined fluorochromes has become instrumental in understanding specific cellular components and complex biological cell structures.
The technique of fluorescence microscopy has become an essential tool in biology and the biomedical sciences, as well as in materials science due to attributes that are not readily available in other contrast modes with traditional optical microscopy. The application of an array of fluorochromes has made it possible to identify cells and sub-microscopic cellular components with a high degree of specificity amid non-fluorescing material. In fact, the fluorescence microscope is capable of revealing the presence of a single molecule. Through the use of multiple fluorescence labeling, different probes can simultaneously identify several target molecules simultaneously. Although the fluorescence microscope cannot provide spatial resolution below the diffraction limit of specific specimen features, the detection of fluorescing molecules below such limits is readily achieved.
A variety of specimens exhibit autofluorescence (without the application of fluorochromes) when they are irradiated, a phenomenon that has been thoroughly exploited in the fields of botany, petrology, and the semiconductor industry. In contrast, the study of animal tissues and pathogens is often complicated with either extremely faint or bright, nonspecific autofluorescence. Of far greater value for the latter studies are added fluorochromes (also termed fluorophores), which are excited by specific wavelengths of irradiating light and emit light of defined and useful intensity. Fluorochromes are stains that attach themselves to visible or sub-visible structures, are often highly specific in their attachment targeting, and have a significant quantum yield (the ratio of photon absorption to emission). The widespread growth in the utilization of fluorescence microscopy is closely linked to the development of new synthetic and naturally occurring fluorophores with known intensity profiles of excitation and emission, along with well-understood biological targets.
Fluorescence microscopes are the tool used to irradiate a sample with a specific set of wavelengths, and optically separate the excitation light from the emitted fluorescence light to obtain an image of the structure being observed. Light sources, internal optics, and detection advances have resulted in the ability to have higher resolution and more complex imaging yielding far more information than was previously available.
The Vortran Laser Stradus VersaLase is one of those advances to help advance fluorescence microscopy capabilities. There are many factors that make this system the right choice for illuminating complex samples. The individual laser design includes patented technology creating an ultra-stable beam in all the necessary components of power stability, polarization stability, and ultra-low optical noise. The unique internal power feedback look of the system forces the output of the laser to not fluctuate in power. The result is a consistent excitation signal for either short term or extremely long-term exposure times. The manufacturing process of the lasers was designed to reduce induced stress caused by standard manufacturing processes. The inherent stress from machining and assembly can produce bi-refringence stress in the optical train resulting in shifting of the beam affecting the sample excitation. Our patented technology and the use of electronic design IP gathered over decades of laser design produces the lowest optical noise beam in the industry.
Regardless of the microscope used, our system has all the necessary connections and compatibility to not only work seamlessly with a new or existing setup, it will perform better than what is typically supplied with a new system. This is due to the fact we are constantly tuning and upgrading our product to produce the absolute best system available today because today is always changing.
