LNNano - Brazilian Nanotechnology National Laboratory

3D Laser Scanning Microscopy – model VK-X200 (Keyence)

3d-lsm-lnnanoThe laser microscope is a device for imaging and measurement that enables sample observation using a large depth of focus and 3D measurement simultaneously. It couples a laser with an optical microscope to create high-resolution images with nanometer Z-axis measurement data. These types of systems are typically known as “laser scanning microscopes”, but can also be referred to by the acronyms “LSM” or “LSCM” (laser scanning confocal microscope).

The 3D VK-X200 LSCM detects the height information based on reflected light intensity from the sample. The lens performs a scanning of the z-axis and, at the same time, the software stores the data to recreate the 3D image. The objective lens is then driven in the Z-axis direction and the scanning process is repeated to obtain the reflected light intensity at the Z-axis position of each point. The system then records the height information and reflected light intensity by assuming the Z-axis focal-point position is located where the reflected light intensity is highest. With this method, a fully focused light intensity image with height information can be captured.

Technical information:
Wavelength – 408nm, violet
Samples dimension:
80mm x 80mm x 28mm

Operational steps

  1. The beam emitted from the laser light source passes through the X-Y scanning optical system and raster across the sample surface.
  2.  The laser light receiving element detects reflected light intensity information only for positions in focus by the confocal optical system.
  3. A fully focused image is created by accumulating in-focus position information in the Z-axis direction.
  4. At the same time, the focal position of the objective lens is recorded to measure the 3D shape of the surface.


How the laser microscope VKX-200 works (Keyence, guidebook for laser scanning microscopes)

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