Toxicologic Pathology

 

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Toxicologic Pathology, Vol. 32, No. 3, 357-363 (2004)
DOI: 10.1080/01926230490431772

Electron Microscopy of Wet Tissues: A Case Study in Renal Pathology

Abraham Nyska

Laboratory of Experimental Pathology, National Institute of Environmental Health Sciences (NIEHS), Research Triangle Park, North Carolina 27709, USA, nyska{at}niehs.nih.gov

Connie A. Cummings

Pathology Associates (PAI) Division of Charles River Laboratories, Inc., Durham, North Carolina 27713, USA

Anya Vainshtein

QuantomiX, Ltd., Nes Ziona 70400, Israel

Jonathan Nadler

Harlan Biotech Israel Ltd., Kiryat Weizmann, Rehovot 76326, Israel

Nathan Ezov

Harlan Biotech Israel Ltd., Kiryat Weizmann, Rehovot 76326, Israel

Yona Grunfeld

Harlan Biotech Israel Ltd., Kiryat Weizmann, Rehovot 76326, Israel

Opher Gileadi

QuantomiX, Ltd., Nes Ziona 70400, Israel

Vered Behar

QuantomiX, Ltd., Nes Ziona 70400, Israel

In this report we introduce wet-tissue scanning electron microscopy, a novel technique for direct imaging of wet tissue samples using backscattered electrons. Samples placed in sealed capsules are imaged through a resilient, electron-transparent membrane. The contrast of the imaged samples may be enhanced by chemical staining. The samples several millimeters thick and imaged without sectioning, makes this technique suitable for rapid analysis of tissue specimens. We applied this technique to D-limonene-induced nephropathy where accumulation of hyaline protein droplets is induced in proximal and distal convoluted tubules of the kidney. Images obtained by scanning electron microscopy of hydrated kidney specimens exhibited superior resolution, contrast, and magnification compared with those obtained by conventional light microscopy of paraffin sections. The electron micrographs can be obtained within an hour of tissue removal, whereas preparation for light microscopy requires at least 1 day. These advantages of the wet scanning electron microscopy technique indicate its potential utility in a wide range of applications in histopathology and toxicology.

Key Words: Scanning electron microscopy • imaging • wet samples • kidney.


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