Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies
- 22 July 2016
- journal article
- Published by Optica Publishing Group in Biomedical Optics Express
- Vol. 7 (8), 3089-3096
- https://doi.org/10.1364/boe.7.003089
Abstract
We present a multiphoton microscopy approach with clearing optimized for pathology evaluation producing image quality comparable to traditional histology. Use of benzyl alcohol/benzyl benzoate with 4',6-diamidino-2-phenylindole and eosin in an optimized imaging setup results in optical sections nearly indistinguishable from traditionally-cut sections. Application to human renal tissue demonstrates diagnostic-level image quality can be maintained through 1 millimeter of tissue. Three dimensional perspectives reveal changes of glomerular capsule cells not evident on single sections. Collagen-derived second harmonic generation can be visualized through entire biopsies. Multiphoton microscopy with clearing has potential for increasing the yield of histologic evaluation of biopsy specimens.Keywords
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