Confocal Microscopy Lab (CLSM)
SINTEF has a Leica TCS 4D confocal laser scanning microscope (clsm). The microscope is located in Oslo. The department has several years of experince using clsm for examining e.g. paper, coated surfaces, and metals.
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| Picture taken from the lab in Oslo. |
Main advantages
The most important advantage of the clsm compared to conventional optical microscopes is the optical sectioning effect which significantly improves the 3D-imaging of thick, transparent specimens.
Furthermore, the clsm has better transverse resolution than conventional optical microscopes, and can measure height differences up to 170 µm.
Applications
The 3D-imaging properties of the clsm makes it ideally suited for studying topography, internal structure, and fluorescence properties of biological objects and materials with dimensions in the sub-mm to mm range. Typical examples of such applications are:
The confocal principle
The basic principle of a confocal microscope is illustrated in the figure below. Light from a laser is focused onto a pinhole. The pinhole is imaged onto the object by an objective lens. Reflected light and/or fluorescence from the object is imaged by the same objective lens onto a detection pinhole via a beamsplitter. A photomultiplier-detector records the light transmitted through the pinhole. In this way, light rays generated from outside the focused region is effectively suppressed by the detection pinhole (optical sectioning-effect). An image of the object is obtained by scanning the light beam in x/y-directions, in raster-like manner. Finally, a 3D-image is obtained by scanning the object in z-direction (along the optical axis) and stacking the images obtained for each z-value.
Some technical data:
Illumination at wavelength=488 nm (blue), wavelength=568 nm (green), and wavelength=647 nm (red), confocal imaging in reflection mode and fluorescence mode, conventional imaging in transmission mode or reflection mode using white light or laser illumination, z-resolution of 350 nm (FWHM) at wavelength=488 nm and NA=1.32, topography of surfaces can be measured with 50-100 nm (relative) accuracy, x/y-resolution of 180 nm (FWHM) at wavelength=488 nm and NA=1.32, maximum imaging range of 2 mm (x), 2 mm (y), and 170 µm (z) per scan series, diffraction limited optics, microscope can be operated as a conventional microscope with video-imaging possibilities.
If you are interested in more information please contact
Odd Løvhaugen
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The above image shows a x-z section through a metallic lacquer. From this image we see the metallic particles lying about 30 mikrometers below the lacquer surface. |
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The above image shows a x-y section in the same metallic lacquer as the previous image. |
