Confocal Laser Scanning Microscopy (CLSM)

Confocal Laser Scanning Microscopy (CLSM)

Technique:
Confocal Laser Scanning Microscopy (CLSM) for materials uses laser light (405 nm wavelength) and an adjustable aperture to optically produce thin (nm to um) focal slices to optically section a sample. The aperture (typically called a pinhole) rejects out-of-focus information, and only in-focus information is detected. By changing the distance between the sample and the objective lens, through finely controlled steps in the Z direction, and by scanning the laser in the X/Y direction, an image stack is created with each pixel of each slice containing intensity information about the investigated surface. The most intense pixels of the stack are combined to form a continuous surface.

The applications of the instrument cover roughness/waviness estimation, and step height measurements. By scanning an area or montaging together many areas, a three-dimensional topographic image of the surface can be produced, and two-dimensional profiles can be isolated from this for analysis.

Instrument:

Zeiss LSM800 For Materials / Zeiss Axio Imager.Z2m upright compound microscope

System Capabilities:

  • Full spectrum LED light for reflective microscopy
  • 405 nm wavelength laser
  • C-Epiplan-Apochrome Objectives: 5x, NA 0.2; 10x, NA 0.4; 20x, NA 0.4 LD; 20x, NA 0.7; 50x, NA 0.55 LD; 50x, NA 0.95
  • Piezo measured motorized stage
  • Brightfield and Circular-DIC reflector cubes
  • TIC filter

Laser confocal microscope image of a printed circuit board used for electrochemical testing.


Selected Applications in Industry:

  • Monitoring of corrosion growth on metals
  • Analysis of fiber texture of paper
  • Measurement of thickness of an ink printed surface
  • Membrane and fabric surface 3D morphology imaging
  • Crater depth measurement of a paint basecoat underneath a clearcoat
  • SIMS sputter rate estimation