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Ultrasound-accelerated Tissue Fixation/Processing Achieves Superior Morphology and Macromolecule Integrity with Storage Stability

Wei-Sing Chu, Qi Liang, Yao Tang, Randy King, Kondi Wong, Maokai Gong, Minqi Wei, Jilan Liu, Shaw-Huey Feng, Shyh-Ching Lo, Jo-Ann Andriko and Marshall Orr

Department of Scientific Laboratories (W-SC,QL,RK,MG,MW,JL), Neuropathology and Ophthalmic Pathology (KW), Infectious and Parasitic Diseases Pathology (S-HF,S-CL), Armed Forces Institute of Pathology, Washington, DC; Department of Pathology, Walter Reed Army Medical Center, Washington, DC (J-AA); Naval Research Laboratory, Washington, DC (MO); and Cancer Center, University of Maryland School of Medicine, Baltimore, Maryland (YT)

Correspondence to: Wei-Sing Chu, MD, Department of Scientific Laboratories, Armed Forces Institute of Pathology, Washington, DC 20306-6000. E-mail: chu{at}afip.osd.mil

We demonstrate that high-frequency and high-intensity ultrasound (US) can be applied to both tissue fixation and tissue processing to complete the conventional overnight formalin-fixation and paraffin-embedding (FFPE) procedures within 1 hr. US-facilitated FFPE retains superior tissue morphology and long-term room temperature storage stability than conventional FFPE. There is less alteration of protein antigenicity after US-FFPE preservation so that rapid immunohistochemical reactions occur with higher sensitivity and intensity, reducing the need for antigen retrieval pretreatment. US-FFPE tissues present storage stability so that room temperature storage up to 7 years does not significantly affect tissue morphology, protein antigenic properties, RNA distribution, localization, and quantitation. In addition, during fixation, tissue displays physical changes that can be monitored and reflected as changes in transmission US signals. As far as we know, this is the first effort to monitor tissue physical changes during fixation. Further study of this phenomenon may provide a method to control and to monitor the level of fixation for quality controls. The mechanism of less alteration of protein antigenicity by US-FFPE was discussed. (J Histochem Cytochem 54:503–513, 2006)

Key Words: ultrasound • formalin fixation • tissue processing • immunohistochemistry • RNA extraction • storage stability

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