How to choose a suitable fluorescence microscope
Fluorescence microscope is a standard microscopic imaging equipment in laboratories and pathology departments, which uses fluorescence characteristics for observation and imaging. It is widely used in various fields such as cell biology, neurobiology, botany, microbiology, pathology, genetics, etc. Fluorescence imaging has the advantages of high sensitivity and specificity, making it very suitable for observing the distribution of specific proteins, organelles, etc. in tissues and cells, studying co localization and interactions, tracking dynamic processes such as changes in ion concentration, and so on.
Selection of Fluorescent Light Source
The commonly used fluorescent light sources currently include mercury lamps, metal halide lamps, and the rapidly developing LED light sources in recent years. The spectrum of fluorescent light sources can be continuous or discontinuous, and the energy varies in different bands. LED light sources are gradually becoming the main light source for fluorescence microscopes due to their relatively narrow spectral bands, more stable energy output, longer lifespan, and greater safety and environmental friendliness.
Confocal microscope
In traditional fluorescence microscopy observation, due to the overlap of fluorescent labeling substances and spontaneous fluorescence structures, they are tightly bound together. However, traditional epifluorescence microscopy objectives not only collect light from the focal plane, but also collect scattered light above and below the focal plane, resulting in greatly reduced image resolution and contrast.
Confocal imaging only detects the portion of light reflected from the focal plane, thus solving the above problem. The light source forms a small and fine spot on the focal plane through a pinhole, and the light emitted from the focal plane is collected by the objective lens. The majority of the fluorescence emitted from points above or below the focal plane of the objective lens cannot converge to the pinhole. Only the fluorescence located on the focal plane and a small portion of defocused fluorescence can pass through the pinhole, while the light beam outside the focal plane converges in front or behind the pinhole plate and is blocked from entering the detector through the pinhole. The detected image is the image from the focal plane, so the final image quality is greatly improved.
Due to the various advantages and practicality of laser scanning confocal microscopy, confocal microscopy is currently an indispensable experimental assistant in the high-precision fields of cell biology, botany, and cell research. At the same time, in future scientific research centers, it will be the most fundamental and core research tool.
