Research Papers: Imaging

Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance

[+] Author Affiliations
Jason Bini

Memorial Sloan-Kettering Cancer Center, Department of Dermatology, New York, New York 10022

Stevens Institute of Technology, Department of Chemistry, Chemical Biology and Biomedical Engineering, Hoboken, New Jersey 07030

James Spain, Kishwer Nehal, Milind Rajadhyaksha

Memorial Sloan-Kettering Cancer Center, Department of Dermatology, New York, New York 10022

Vikki Hazelwood

Stevens Institute of Technology, Department of Chemistry, Chemical Biology and Biomedical Engineering, Hoboken, New Jersey 07030

Charles DiMarzio

Northeastern University, Department of Electrical and Computer Engineering, Boston, Massachusetts 02115

Northeastern University, Department of Mechanical and Industrial Engineering, Boston, Massachusetts 02115

J. Biomed. Opt. 16(7), 076008 (July 14, 2011). doi:10.1117/1.3596742
History: Received February 28, 2011; Revised April 25, 2011; Accepted May 13, 2011; Published July 14, 2011; Online July 14, 2011
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Confocal mosaicing microscopy enables rapid imaging of large areas of fresh tissue, without the processing that is necessary for conventional histology. Mosaicing may offer a means to perform rapid histology at the bedside. A possible barrier toward clinical acceptance is that the mosaics are based on a single mode of grayscale contrast and appear black and white, whereas histology is based on two stains (hematoxylin for nuclei, eosin for cellular cytoplasm and dermis) and appears purple and pink. Toward addressing this barrier, we report advances in digital staining: fluorescence mosaics that show only nuclei, are digitally stained purple and overlaid on reflectance mosaics, which show only cellular cytoplasm and dermis, and are digitally stained pink. With digital staining, the appearance of confocal mosaics mimics the appearance of histology. Using multispectral analysis and color matching functions, red, green, and blue (RGB) components of hematoxylin and eosin stains in tissue were determined. The resulting RGB components were then applied in a linear algorithm to transform fluorescence and reflectance contrast in confocal mosaics to the absorbance contrast seen in pathology. Optimization of staining with acridine orange showed improved quality of digitally stained mosaics, with good correlation to the corresponding histology.

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© 2011 Society of Photo-Optical Instrumentation Engineers (SPIE)

Citation

Jason Bini ; James Spain ; Kishwer Nehal ; Vikki Hazelwood ; Charles DiMarzio, et al.
"Confocal mosaicing microscopy of human skin ex vivo: spectral analysis for digital staining to simulate histology-like appearance", J. Biomed. Opt. 16(7), 076008 (July 14, 2011). ; http://dx.doi.org/10.1117/1.3596742


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