A three-dimensional calibration device for the confocal microscope


Jensen, K. E. ; Weitz, D. A. ; Spaepen, F. A three-dimensional calibration device for the confocal microscope. Review of Scientific Instruments 2013, 84. Copy at http://www.tinyurl.com/yxk3z8ss
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Modern confocal microscopes enable high-precision measurement in three dimensions by collecting stacks of 2D (x-y) images that can be assembled digitally into a 3D image. It is difficult, however, to ensure position accuracy, particularly along the optical (z) axis where scanning is performed by a different physical mechanism than in x-y. We describe a simple device to calibrate simultaneously the x, y, and z pixel-to-micrometer conversion factors for a confocal microscope. By taking a known 2D pattern and positioning it at a precise angle with respect to the microscope axes, we created a 3D reference standard. The device is straightforward to construct and easy to use. (C) 2013 American Institute of Physics. [http://dx.doi.org/10.1063/1.4776672]

Last updated on 08/08/2014