title: Multiphoton microscopy with clearing for three dimensional histology of kidney biopsies.
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.
authors: Dept of Laboratory Medicine, Yale School of Medicine, New Haven, CT 06510, USA.
authors: Eben
authors: E
authors: Olson
authors: Applikate Technologies, Weston, CT 06883, USA.
authors: Michael J
authors: MJ
authors: Levene
authors: Dept of Laboratory Medicine, Yale School of Medicine, New Haven, CT 06510, USA.
authors: Richard
authors: R
authors: Torres
keywords: (180.0180) Microscopy; (180.4315) Nonlinear microscopy; (180.6900) Three-dimensional microscopy