Skin Imaging With Reflectance Confocal Microscopy

Abstract

Confocal microscopy is a new imaging modality for noninvasive real-time tissue imaging with high resolution and contrast comparable with conventional histology. Application of this technology to skin imaging during the last decade has been an exciting advance in dermatology, allowing a virtual widow into living skin without the need for a conventional biopsy or histologic processing of tissue. High-resolution noninvasive skin imaging with confocal microscopy has potential broad applications in the clinical and research arenas, including differentiating between benign and malignant skin lesions, tumor margin mapping, monitoring response to medical or surgical treatments, and pathophysiologic study of inflammatory processes.

In vivo imaging of cell nuclei by photoacoustic microscopy without staining

“…The high image contrast results from a difference between strong light absorption of nucleic acids and weak absorption of protein, which reaches the maximum at a wavelength of 250 nm. In addition to UV-PAM, several other modern optical microscopy technologies have been explored for in vivo imaging of unstained cell nuclei, such as reflectance confocal microscopy [8][9][10][11], multiphoton microscopy [12][13][14], and third-harmonic generation microscopy [15,16], However, reflectance confocal microscopy is difficult to provide specific image contrast for nuclei [8], multiphoton microscopy produces in vivo images of cell nuclei with negative contrast [13], and third harmonic generation microscopy generates nuclear images with low contrast [15].…”

Progress in dermatology

“…The article by Meschieri et al [9] reviews the value of in-vivo reflectance confocal microscopy as a tool in cutaneous oncology. One of his co-authors, Millind Rajadhyaksha has been crucial in developing this technology and bringing it to practical application [10][11][12][13][14]. As the major technical limitation of this method is the image depth of up to 300 μm, its main role may be in early detection of skin malignancies, specifically in directing necessary biopsies and avoiding unnecessary ones.…”

Determination of Melanoma Lateral and Depth Margins: Potential for Treatment Planning and Five-Year Survival Rate

“…The basic premise of SCM is the selective collection of light from a specific plane in tissue through a pinhole-sized aperture which allows for light collection from the single in-focus plane and the rejection of light from all out-of-focus planes (Nehal et al, 2008). SCM has been recently employed in CMM diagnosis (Gerger et al, 2005;Marghoob & Halpern, 2005), preoperative and intraoperative margin assessment (Busam et al, 2001), and followup for response to medical treatment (Ahmed & Berth-Jones, 2000;Cornejo et al, 2000;Langley et al, 2006;Tannous et al, 2000Tannous et al, , 2002.…”

“…Commercial SCM instruments have been developed that image with lateral resolution of 0.5 to 1.0 μm and an optical sectioning thickness of 1.0 to 5.0 μm, to a depth of 200 to 300 μm in human skin (depth of papillary dermis). The spatial resolution of SCM is determined by the pinhole size while imaging depth is limited by the laser wavelength (with a 488 nm laser imaging 50-100 μm into skin (Gareau et al, 2007) and longer wavelengths lasers able to image at depths of up to 300 μm (Gonzalez & Gilaberte-Calzada, 2008;Marghoob et al, 2003;Nehal et al, 2008), providing images of the basement membrane down into the papillary dermis. SCM with an 830 nm light source is ideal for detecting CMM because melanin serves as an endogenous contrast agent.…”