Laser Scanning Confocal Microscope Diagram

A thick 16 micrometer section of fluorescently stained.

Laser scanning confocal microscope diagram.

Confocal imaging confocal image of convallaria lily of the valley. The pinhole aperture is 2 airy units. 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. 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.

Confocal laser scanning microscopy principles microscopy from carl zeiss optical image formation. Confocal microscopy can be considered a bridge between these two classical methodologies. Capturing multiple two dimensional images at different depths in a sample enables the. Clsm combines high resolution optical imaging with depth selectivity which allows us to do optical sectioning.

Laser excitation wavelength is 561nm supercontinuum laser. Illustrated in figure 1 are a series of images that compare selected viewfields in traditional widefield and laser scanning confocal fluorescence microscopy. The classical abbe equation describes the relationship between spatial resolution the excitation wavelength and the numerical aperture of the. Fluorescent microscopy not only makes our images look good it also allows us to gain a better understanding of cells structures and tissue.

Image information is gathered point. Among various devices confocal laser scanning microscopy clsm is considered to be a valuable tool in dentistry research. 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. 3 non confocal top and confocal bottom image of a double.

All laser scanning confocal microscope designs are centered around a conventional upright or inverted research level optical microscope. With confocal laser scanning microscopy clsm we can find out even more. The fluorescence is collected through a bandpass filter centered at 600 nm with 37 nm bandpass. The objective is an olympus 20x water immersion na 0 54.

Two photon excitation microscopy tpef or 2pef is a fluorescence imaging technique that allows imaging of living tissue up to about one millimeter in thickness. Diagram 5 excitation photon flux at different laser powers diagram 6 excited state saturation behavior absorb ed photons.