Original Article

Journal of Investigative Dermatology (1999) 113, 293–303; doi:10.1046/j.1523-1747.1999.00690.x

In Vivo Confocal Scanning Laser Microscopy of Human Skin II: Advances in Instrumentation and Comparison With Histology1

Milind Rajadhyaksha*,, Salvador González, James M Zavislan*, R Rox Anderson and Robert H Webb

  1. *Lucid Inc., Henrietta, New York, U.S.A.
  2. Wellman Laboratories of Photomedicine, Department of Dermatology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, U.S.A.

Correspondence: Dr Milind Rajadhyaksha, MGH-Wellman Labs, Bartlett Hall Ext. 630, Boston, MA 02114, U.S.A. E-mail: rajadmil@helix.mgh.harvard.edu

1The authors have declared conflict of interest.

Received 28 August 1998; Revised 19 May 1999; Accepted 25 May 1999.

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Abstract

In 1995, we reported the construction of a video-rate scanning laser confocal microscope for imaging human skin in vivo. Since then, we have improved the resolution, contrast, depth of imaging, and field of view. Confocal images of human skin are shown with experimentally measured lateral resolution 0.5–1.0 mum and axial resolution (section thickness) 3–5 mum at near-infrared wavelengths of 830 nm and 1064 nm; this resolution compares well to that of histology which is based on typically 5 mum thin sections. Imaging is possible to maximum depth of 350 mum over field of view of 160–800 mum. A mechanical skin-contact device was developed to laterally stabilize the imaging site to within plusminus 25 mum in the presence of subject motion. Based on these results, we built a small, portable, and robust confocal microscope that is capable of imaging normal and abnormal skin morphology and dynamic processes in vivo, in both laboratory and clinical settings. We report advances in confocal microscope instrumentation and methods, an optimum range of parameters, improved images of normal human skin, and comparison of confocal images with histology.

Keywords:

dermatology, morphology, non-invasive imaging, optical imaging

Abbreviations:

CM, confocal microscope; NA, numerical aperture

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