Laser Scanning Confocal Microscope Diagram

Confocal laser scanning microscopy principles microscopy from carl zeiss optical image formation.
Laser scanning confocal microscope diagram. Its applications include research on caries response of hard and soft. Clsm combines high resolution optical imaging with depth selectivity which allows us to do optical sectioning. Laser excitation wavelength is 561nm supercontinuum laser. Among various devices confocal laser scanning microscopy clsm is considered to be a valuable tool in dentistry research.
Unlike traditional fluorescence microscopy in which the excitation wavelength is shorter than the emission wavelength two photon excitation requires simultaneous excitation by two photons with longer wavelength than the emitted light. The point to point lateral x y resolution of a spinning disk confocal microscope is essentially the same as that in laser scanning confocal or widefield fluorescence microscopy see figure 4 b. The classical abbe equation describes the relationship between spatial resolution the excitation wavelength and the numerical aperture of the. Capturing multiple two dimensional images at different depths in a sample enables the.
With confocal laser scanning microscopy clsm we can find out even more. However instead of the standard tungsten halogen or mercury arc discharge lamp one or more laser systems are used as a light source to excite fluorophores in the specimen. Confocal microscopy can be considered a bridge between these two classical methodologies. A thick 16 micrometer section of fluorescently stained.
The fluorescence is collected through a bandpass filter centered at 600 nm with 37 nm bandpass. The pinhole aperture is 2 airy units. Fluorescent microscopy not only makes our images look good it also allows us to gain a better understanding of cells structures and tissue. All laser scanning confocal microscope designs are centered around a conventional upright or inverted research level optical microscope.
Illustrated in figure 1 are a series of images that compare selected viewfields in traditional widefield and laser scanning confocal fluorescence microscopy. Diagram 5 excitation photon flux at different laser powers diagram 6 excited state saturation behavior absorb ed photons. 3 non confocal top and confocal bottom image of a double. This means that we can view visual sections of tiny structures that.
Confocal imaging confocal image of convallaria lily of the valley. The objective is an olympus 20x water immersion na 0 54.