[54] DUAL PURPOSE TISSUE FIXATIVE
[76] Inventor: Hyman C. Birnboim, 1552 Featherston Drive, Ottawa, Ontario, Canada, K1H 6P2
[21] Appl. No.: 08/708,056 [22] Filed: Aug. 30, 1996
Related U.S. Application Data
[63] Continuation of application No. 08/231,247, Apr. 22, 1994, abandoned.
[51] Int. CI.6 C12Q 1/08; C12Q 1/00
[52] U.S. CI 435/40.5; 435/40.52; 435/1.1;
424/75; 436/8; 436/17; 436/18
[58] Field of Search 435/40.5, 40.52,
435/240.1, 1.1; 436/8, 17, 18; 424/75
[56] References Cited
U.S. PATENT DOCUMENTS
4,826,691 5/1989 Prochnow 426/1
4,857,300 8/1989 Maksem et al 424/3
4,888,278 12/1989 Singer et al 435/6
5,196,182 3/1993 Ryan 424/3
5,357,977 10/1994 Michels 128/758
5,389,549 2/1995 Hamaguchi et al 436/10
5,405,606 4/1995 Campbell et al 424/75
FOREIGN PATENT DOCUMENTS
1-140064 1/1989 Japan .
OTHER PUBLICATIONS
Eidus et al "A New Fixative for Molecular Biology and Diagnostic Pathology: Approximating a Universal Fixative" FASEB J 8(4-5) 1994 p. A391.
J. Histochem. & Cytochem. McAllister et al. vol. 33, No. 10 pp. 1026-1032 1985. Comparative Usefullness of Tissue Fixatives for In Situ Viral.
Exp. Hematol. Bramwell et al, 16:730-732, 1988. The Effects of Fixative Type and Fixation Time on the Quantity and Quality of Extractable DNA.
Am. J. Clin. Pathol. Greer et al. 95:117-124. 1991. PCR Amplification from Paraffin-Embedded Tissues.
Anatomic Pathology, Esteban et al. Apr. 1991, pp. 460-465; Effects of Various Fixatives and Fixation Conditions on DNA Ploidy Analysis.
Am. J. Hum. Genet. Barker et al. 39:661-668, 1986. High Molecular Weight DNA from Fixed Cytogenetic Preparations.
Anal. Biochem. Smith, et al. 160:135-138,1987. Extraction of Cellular DNA from Human Cells and Tissues fixed in Ethanol.
J. Histochem & Cytochem. Ben-ezra et al.1991 vol. 39. No. 3 pp. 351-354 Effect of Fixation on the Amplification of Nucleic acids from Paraffin.
J. Histochem. & Cytochem. WEiss et al. 1991 vol. 39 No. 9 pp. 1237-1242 Effects of Different Fixatives on Detection of Nucleic Acids from.
Arch. Pathol. Lab. Med., Bostwick, D.G., vol. 118, pp. 298-302 1994. Establishment of the Formalin-Free Surgical Pathology Laboratory.
Primary Examiner—Paula K. Hutzell
Assistant Examiner—Heather A. Bakalyar
Attorney, Agent, or Firm—Lorusso & Loud
[57] ABSTRACT
A dual purpose tissue fixative is described. The fixative comprises a cross-linking aldehyde, alcohol, a chelating agent and a non-amine buffer. More particularly, the tissue fixative comprises 0.1-2% w/v formaldehyde, 45-90% w/w alcohol, 1-10 mM of a chelating agent and 1-50 mM of a non-nitrogen buffer. The tissue fixative permits the recovery of high molecular weight DNA and RNA from the tissue sample for molecular genetic analysis. The tissue fixative also preserves the morphology and immunogenicity of the tissue allowing for pathology analysis. The tissue fixative is compared to 95% alcohol and a standard fixative as 10% BNF.
8 Claims, 6 Drawing Sheets
