Working principle of laser confocal microscope
Working principle of laser confocal microscope
Olympus laser confocal microscope is a detector that can acquire signals. Its working principle is that a point light source will be imaged as an enlarged spot called "Airy disk" after passing through a fluorescence microscope. In a standard white-light interferometric confocal microscope, light emitted outside the focal plane is blocked by a pinhole whose size determines how much of the Airy disk can enter the detector. The smaller the pinhole, the sharper the resulting image, and the dimmer the image because most of the light is lost. The smaller the aperture pinhole, the better the resolution, but—again—the more light signal is lost. Olympus laser confocal microscope can provide a color CCD image and a laser scanning confocal image at the same time. Olympus confocal laser microscopes are capable of height measurement through a confocal setup of one light source, one sample, and one detector. When the sample is located at the focal plane of the objective lens and the laser light reflected from the sample surface is focused into the confocal aperture, the photodetector will receive the signal from the sample. When the sample is in the out-of-focus position, the confocal aperture does not receive the laser signal, so only the in-focus signal is collected. This feature can realize the optical sectioning function of the Olympus laser confocal microscope.
Advantages of Confocal Laser Microscopy
1. Using laser light as the light source, after the corresponding fluorescent probes are marked, the sample is scanned point by point to obtain two-dimensional optical cross-sectional images layer by layer. It has the function of "cell CT" and can be supported by computer three-dimensional reconstruction software to obtain three-dimensional images, which can be rotated at any angle to observe the three-dimensional shape and spatial relationship of cells and tissues;
2. It can observe living cells and tissues without damage, and dynamically measure the physiological information of living cells such as Ca ion concentration and pH value in cells;
3. It can measure the fluidity of cell membrane, intercellular communication, cell fusion, cytoskeleton elasticity, etc., and can be used as a "light knife" to complete intracellular "surgery". This technology enables in situ dynamic and quantitative observation and measurement of living cells and tissues.
