AJTR Copyright © 2009-All rights reserved. Published by e-Century Publishing Corporation, Madison, WI 53711
Am J Transl Res 2012;4(2):198-205

Original Article
High quality DNA obtained with an automated DNA extraction method
with 70+ year old formalin-fixed celloidin-embedded (FFCE) blocks
from the indiana medical history museum

Erin E Niland, Audrey McGuire, Mary H Cox, George E Sandusky

Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, Van Nuys Medical
Science Building, 635 Barnhill Drive, Room A128, Indianapolis, IN, 46202, USA; Indiana Clinical Translational
Science Institute Department of Medicine, 970 W. Walnut St. Indianapolis, IN 46202 USA

Received January 5, 2012; accepted January 19, 2012; April 10, 2012; Published April 30, 2012

Abstract: DNA and RNA have been used as markers of tissue quality and integrity throughout the last few
decades. In this research study, genomic quality DNA of kidney, liver, heart, lung, spleen, and brain were analyzed
in tissues from post-mortem patients and surgical cancer cases spanning the past century. DNA extraction was
performed on over 180 samples from: 70+ year old formalin-fixed celloidin-embedded (FFCE) tissues, formalin-
fixed paraffin-embedded (FFPE) tissue samples from surgical cases and post-mortem cases from the 1970’s,
1980’s, 1990’s, and 2000’s, tissues fixed in 10% neutral buffered formalin/stored in 70% ethanol from the 1990’s,
70+ year old tissues fixed in unbuffered formalin of various concentrations, and fresh tissue as a control. To
extract DNA from FFCE samples and ethanol-soaked samples, a modified standard operating procedure was
used in which all tissues were homogenized, digested with a proteinase K solution for a long period of time (24-
48 hours), and DNA was extracted using the Autogen Flexstar automated extraction machine. To extract DNA from
FFPE, all tissues were soaked in xylene to remove the paraffin from the tissue prior to digestion, and FFPE
tissues were not homogenized. The results were as follows: celloidin-embedded and paraffin-embedded tissues
yielded the highest DNA concentration and greatest DNA quality, while the formalin in various concentrations, and
long term formalin/ethanol-stored tissue yielded both the lowest DNA concentration and quality of the tissues
tested. The average DNA yield for the various fixatives was: 367.77 μg/mL FFCE, 590.7 μg/mL FFPE, 53.74 μg/mL
formalin-fixed/70% ethanol-stored and 33.2 μg/mL unbuffered formalin tissues. The average OD readings for
FFCE, FFPE, formalin-fixed/70% ethanol-stored tissues, and tissues fixed in unbuffered formalin were 1.86, 1.87,
1.43, and 1.48 respectively. The results show that usable DNA can be extracted from tissue fixed in formalin and
embedded in celloidin or paraffin from the early 1900’s to present, and may be amplified through PCR and used
for clinical and experimental studies. (AJTR1201001).

Keywords: DNA, celloidin, museum specimens

Address all correspondence to:
Dr. Sandusky GE
IU School of Medicine
Department of Pathology, Medical Science Rm. 128
635 Barnhill Dr., Indianapolis, IN 46202, USA.
Tel: 317-278-2304; Fax: 317-278-2018
E-mail: gsandusk@iupui.edu