Abstract
Several sequential passages are involved in the pre-analytical handling of surgical specimens from resection in the surgical theater to paraffin-embedding and storage. Each passage is highly critical and can significantly affect the preservation of morphology, antigens, and nucleic acids. Some key points in this process are still undefined and are subject to high variability among hospitals. High quality and standardization are demanded and pathologists should therefore work to comply with all novel clinical requests (such as genomic and antigenic testing for targeted molecular therapies). Under-vacuum sealing of surgical pieces can be a safe and reliable alternative to storage in large formalin-filled boxes; it prevents dehydration and favors cooling by removing air. Moreover, it implements tissue banking and preservation of nucleic acids. After transport of specimens to pathological anatomy laboratories, the next passage, fixation, has been the object of several attempt to find alternatives to formalin. However, none of the substitutes proved successful, and formalin fixation is still considered the gold standard for preservation of morphology and antigens. RNA has instead been found to be heavily affected by degradation and fragmentation in formalin-fixed tissues. Based on the hypothesis that RNA degradation would be inhibited by maintaining a low temperature, a protocol based on processing tissues with formalin at low temperature (cold fixation) was evaluated and proved useful in obtaining a reduction in RNA fragmentation. Finally, the problem of storage is discussed, in order to find ways to guarantee feasibility of molecular analyses even years after the original diagnosis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ruijter E, van de Kaa C, Aalders T et al (1998) Heterogeneous expression of E-cadherin and p53 in prostate cancer: clinical implications. BIOMED-II markers for prostate cancer study group. Mod Pathol 11(3):276–281
Medeiros F, Rigl CT, Anderson GG et al (2007) Tissue handling for genome-wide expression analysis: a review of the issues, evidence, and opportunities. Arch Pathol Lab Med 131(12):1805–1816
Chung JY, Braunschweig T, Williams R et al (2008) Factors in tissue handling and processing that impact RNA obtained from formalin-fixed, paraffin-embedded tissue. J Histochem Cytochem 56:1033–1042
Bray SE, Paulin FE, Fong SC et al (2010) Gene expression in colorectal neoplasia: modifications induced by tissue ischaemic time and tissue handling protocol. Histopathology 56:240–250
Hammond ME, Hayes DF, Dowsett M et al (2010) American Society of Clinical Oncology and College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Clin Oncol 28(16):2784–2795
Wolff AC, Hammond ME, Hicks DG et al (2013) American Society of Clinical Oncology; College of American Pathologists. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology and College of American Pathologists clinical practice guideline update. J Clin Oncol 31(31):3997–4013
Portier BP, Wang Z, Downs-Kelly E et al (2013) Delay to formalin fixation ‘cold ischemia time’: effect on ERBB2 detection by in-situ hybridization and immunohistochemistry. Mod Pathol 26(1):1–9
Cancer IARC (2006) Monographs on the evaluation of carcinogenic risk to humans. IARC, Lyon
Vaughan WTBH (1939) The practice of allergy. Maryland CV Mosby, Rockville, p 677
Hendrick DJ, Lane DJ (1975) Formalin asthma in hospital staff. Br Med J 1(5958):607–608
Beane Freeman LE, Blair A, Lubin JH et al (2009) Mortality from lymphohematopoietic malignancies among workers in formaldehyde industries: the National Cancer Institute Cohort. J Natl Cancer Inst 101:751–761
Hall A, Harrington JM, Aw TC (1991) Mortality study of British pathologists. Am J Ind Med 20(1):83–89
Bussolati G, Chiusa L, Cimino A et al (2008) Tissue transfer to pathology labs: under vacuum is the safe alternative to formalin. Virchows Arch 452:229–231
Di Novi C, Minniti D, Barbaro S et al (2010) Vacuum-based preservation of surgical specimens: an environmentally-safe step towards a formalin-free hospital. Sci Total Environ 408:3092–3095
Kristensen T, Engvad B, Nielsen O et al (2011) Vacuum sealing and cooling as methods to preserve surgical specimens. Appl Immunohistochem Mol Morphol 19:460–469
Annaratone L, Marchiò C, Russo R et al (2013) A collection of primary tissue cultures of tumors from vacuum packed and cooled surgical specimens: a feasibility study. PLoS ONE 8:e75193
Comanescu M, Annaratone L, D’Armento G et al (2012) Critical steps in tissue processing in histopathology. Recent Pat DNA Gene Sequences 6:22–32
Comanescu M, Arsene D, Ardeleanu C et al (2012) The mandate for a proper preservation in histopathological tissues. Rom J Morphol Embryol 53:233–242
Moggio A, D’Armiento G, Bussolati B (2012) Efficient stem cell isolation from under vacuum preserved tissue samples. Organogenesis 8(3):71–75
Groenen PJ, Blokx WA, Diepenbroek C et al (2011) Preparing pathology for personalized medicine: possibilities for improvement of the pre-analytical phase. Histopathology 59(1):1–7
Stanta G, Mucelli SP, Petrera F et al (2006) A novel fixative improves opportunities of nucleic acids and proteomic analysis in human archive’s tissues. Diagn Mol Pathol 15(2):115–123
Nietner T, Jarutat T, Mertens A (2012) Systematic comparison of tissue fixation with alternative fixatives to conventional tissue fixation with buffered formalin in a xenograft-based model. Virchows Arch 461(3):259–269
Moelans CB, Oostenrijk D, Moons MJ et al (2011) Formaldehyde substitute fixatives: effects on nucleic acid preservation. J Clin Pathol 64:960–967
Arzt L, Kothmaier H, Quehenberger F et al (2011) Evaluation of formalin-free tissue fixation for RNA and microRNA studies. Exp Mol Pathol 91:490–495
Denouel A, Boissiere-Michot F, Rochaix P et al (2011) An alternative fixative to formalin fixation for molecular applications: the RCL2((R))-CS100 approach. Methods Mol Biol 724:297–307
Goldstein NS, Hewitt SM, Taylor CR et al (2007) Recommendations for improved standardization of immunohistochemistry. Appl Immunohistochem Mol Morphol 15:124–133
Dabbs DJ (2008) Immunohistochemical protocols: back to the future. Am J Clin Pathol 129:355–356
Goldstein NS, Ferkowicz M, Odish E et al (2003) Minimum formalin fixation time for consistent estrogen receptor immunohistochemical staining of invasive breast carcinoma. Am J Clin Pathol 120(1):86–92
Lewis F, Maughan NJ, Smith V et al (2001) Unlocking the archive–gene expression in paraffin-embedded tissue. J Pathol 195:66–71
Fox CH, Johnson FB, Whiting J et al (1985) Formaldehyde fixation. J Histochem Cytochem 33(8):845–853
Stanta G, Schneider C (1991) RNA extracted from paraffin-embedded human tissues is amenable to analysis by PCR amplification. Biotechniques 11:304, 306, 308
Helander KG (1994) Kinetic studies of formaldehyde binding in tissue. Biotech Histochem 69:177–179
Sompuram SR, Vani K, Bogen SA (2006) A molecular model of antigen retrieval using a peptide array. Am J Clin Pathol 125:91–98
Zou N, Liang Q, He H et al (2011) Ultrasound-facilitated formalin fixation of biological specimens. Biotech Histochem 86(6):413–420
Hewitt SM, Lewis FA, Cao Y et al (2008) Tissue handling and specimen preparation in surgical pathology: issues concerning the recovery of nucleic acids from formalin-fixed, paraffin-embedded tissue. Arch Pathol Lab Med 132(12):1929–1935
Durgun-Yücel B, Hopwood D, Yücel AH (1996) The effects of mercaptoethanol-formaldehyde on tissue fixation and protein retention. Histochem J 28(5):375–383
Buesa RJ, Peshkov MV (2012) How much formalin is enough to fix tissues? Ann Diagn Pathol 16:202–209
Dotti I, Bonin S, Basili G et al (2010) Effects of formalin, methacarn, and fineFIX fixatives on RNA preservation. Diagn Mol Pathol 19:112–122
van Maldegem F, de Wit M, Morsink F et al (2008) Effects of processing delay, formalin fixation, and immunohistochemistry on RNA recovery from formalin-fixed paraffin-embedded tissue sections. Diagn Mol Pathol 17:51–58
Paska C, Bogi K, Szilak L et al (2004) Effect of formalin, acetone, and RNAlater fixatives on tissue preservation and different size amplicons by real-time PCR from paraffin-embedded tissue. Diagn Mol Pathol 13:234–240
Masuda N, Ohnishi T, Kawamoto S et al (1999) Analysis of chemical modification of RNA from formalin-fixed samples and optimization of molecular biology applications for such samples. Nucleic Acid Res 27:4436–4443
Bussolati G, Annaratone L, Medico E et al (2011) Formalin fixation at low temperature better preserves nucleic Acid integrity. PLoS ONE 6:e21043-e
Xie R, Chung JY, Ylaya K et al (2011) Factors influencing the degradation of archival formalin-fixed paraffin-embedded tissue sections. J Histochem Cytochem 59(4):356–365
von Ahlfen S, Missel A, Bendrat K et al (2007) Determinants of RNA quality from FFPE samples. PLoS ONE 2(12):e1261
Acknowledgments
This work was supported by Ricerca Sanitaria Finalizzata RF-2010-2310674.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer International Publishing Switzerland
About this chapter
Cite this chapter
Bussolati, G., Annaratone, L., Maletta, F. (2015). The Pre-analytical Phase in Surgical Pathology. In: Dietel, M., Wittekind, C., Bussolati, G., von Winterfeld, M. (eds) Pre-Analytics of Pathological Specimens in Oncology. Recent Results in Cancer Research, vol 199. Springer, Cham. https://doi.org/10.1007/978-3-319-13957-9_1
Download citation
DOI: https://doi.org/10.1007/978-3-319-13957-9_1
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-13956-2
Online ISBN: 978-3-319-13957-9
eBook Packages: MedicineMedicine (R0)