title: Note: A three-dimensional calibration device for the confocal microscope.
abstract: 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.
authors: Department of Physics, Harvard University, Cambridge, Massachusetts 02138, USA. jensen@physics.harvard.edu
authors: K E
authors: KE
authors: Jensen
authors: D A
authors: DA
authors: Weitz
authors: F
authors: F
authors: Spaepen
mesh_terms: D002138:Calibration; D004867:Equipment Design; D021621:Imaging, Three-Dimensional; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000592:standards; D012015:Reference Standards