Ultrasound-accelerated tissue fixation/processing achieves superior morphology and macromolecule integrity with storage stability.
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.
Chu
Wei-Sing
WS
Department of Scientific Laboratories, Armed Forces Institute of Pathology, Washington, DC 20306-6000, USA. chu@afip.osd.mil
Liang
Qi
Q
Tang
Yao
Y
King
Randy
R
Wong
Kondi
K
Gong
Maokai
M
Wei
Minqi
M
Liu
Jilan
J
Feng
Shaw-Huey
SH
Lo
Shyh-Ching
SC
Andriko
Jo-Ann
JA
Orr
Marshall
M
D001344:Autopsy; D001706:Biopsy; D015153:Blotting, Western; D017252:CD3 Complex / Q000032:analysis; D018956:CD5 Antigens / Q000032:analysis; D004247:DNA / Q000032:analysis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D017403:In Situ Hybridization; D007633:Keratins / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D013048:Specimen Handling; D013696:Temperature; D013997:Time Factors; D016707:Tissue Fixation; D014465:Ultrasonics
D017252:CD3 Complex; D018956:CD5 Antigens; C081876:CKAP4 protein, human; D005404:Fixatives; D008565:Membrane Proteins; D012333:RNA, Messenger; D005557:Formaldehyde; D012313:RNA; D007633:Keratins; D004247:DNA