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CC BY-NC-ND 4.0 · Journal of Morphological Sciences 2018; 35(03): 173-176
DOI: 10.1055/s-0038-1673611
Short Communication
Thieme Revinter Publicações Ltda Rio de Janeiro, Brazil

Anatomopathological Assessment of the Diaphragm in Formalin-Fixed, Paraffin-Embedded Sections

Avaliação anatomopatológica do diafragma em cortes fixados em formalina e incorporados em parafina
Ricardo Aparecido Baptista Nucci
1   Department of Pathology, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
2   Postgraduate Pathology program, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
,
Wilson Jacob-Filho
2   Postgraduate Pathology program, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
3   Department of Geriatrics, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
,
Alexandre Leopold Busse
2   Postgraduate Pathology program, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
3   Department of Geriatrics, Faculty of Medicine, Universidade de São Paulo, São Paulo, SP, Brazil
,
Laura Beatriz Mesiano Maifrino
4   Laboratory of Morphoquantitative Studies and Immunohistochemistry, Department of Physical Education, Universidade São Judas Tadeu, São Paulo, SP, Brazil
5   Postgraduate Physical Education program, Universidade São Judas Tadeu, São Paulo, SP, Brazil
,
Romeu Rodrigues de Souza
6   Department of Anatomy, Institute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil
7   Postgraduate Aging Sciences program, Universidade São Judas Tadeu, São Paulo, SP, Brazil
› Author Affiliations
Further Information

Publication History

12 February 2018

09 August 2018

Publication Date:
11 October 2018 (online)

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Abstract

Introduction The analysis of frozen muscle biopsies has become a routine method in the evaluation of muscle structure in health and disease. However, the technique for frozen muscle specimens is not widely available in countries with limited medical facilities. The present study aimed to elucidate a reproducible formalin-fixed and paraffin-embedded (FFPE) method for this type of analysis in postmortem muscles.

Methods Diaphragm muscle was obtained within 1 hour of sudden death. Diaphragm strips were washed in saline solution, fixed in 10% formalin, frozen at 4°C in a refrigerator, and stored for 24 hours. Then, the tissue samples were processed into paraffin-embedded blocks. Transversal sections were cut from each paraffin block and stained with hematoxylin and eosin, Picrosirius red, Verhoeff-Van Gieson, and Congo red for the qualitative analysis.

Results Our analysis indicated a well-preserved muscle.

Conclusion In summary, we demonstrate a simple technique for a reproducible FFPE method in postmortem muscle tissues.

Keywords

anatomy - muscle - necropsy - pathology
 

  • References

  • 1 Poole DC, Sexton WL, Farkas GA, Powers SK, Reid MB. Diaphragm structure and function in health and disease. Med Sci Sports Exerc 1997; 29 (06) 738-754
    CrossrefPubMedGoogle Scholar
  • 2 Sinderby C, Beck J, Spahija J, Weinberg J, Grassino A. Voluntary activation of the human diaphragm in health and disease. J Appl Physiol (1985) 1998; 85 (06) 2146-2158
    CrossrefPubMedGoogle Scholar
  • 3 Topeli A, Laghi F, Tobin MJ. The voluntary drive to breathe is not decreased in hypercapnic patients with severe COPD. Eur Respir J 2001; 18 (01) 53-60
    CrossrefPubMedGoogle Scholar
  • 4 Dubowitz V, Oldfors A, Sewry CA. Muscle Biopsy E-Book: A Practical Approach. Elsevier Health Sciences; 2013
    Google Scholar
  • 5 Eriksson O, Eriksson A, Ringqvist M, Thornell LE. The reliability of histochemical fibre typing of human necropsy muscles. Histochemistry 1980; 65 (03) 193-205
    CrossrefPubMedGoogle Scholar
  • 6 Suriyonplengsaeng C, Dejthevaporn C, Khongkhatithum C. , et al. Immunohistochemistry of sarcolemmal membrane-associated proteins in formalin-fixed and paraffin-embedded skeletal muscle tissue: a promising tool for the diagnostic evaluation of common muscular dystrophies. Diagn Pathol 2017; 12 (01) 19
    PubMedGoogle Scholar
  • 7 Farfel JM, Nitrini R, Suemoto CK. , et al; Brazilian Aging Brain Study Group. Very low levels of education and cognitive reserve: a clinicopathologic study. Neurology 2013; 81 (07) 650-657
    CrossrefPubMedGoogle Scholar
  • 8 Suemoto CK, Ferretti-Rebustini RE, Rodriguez RD. , et al. Neuropathological diagnoses and clinical correlates in older adults in Brazil: A cross-sectional study. PLoS Med 2017; 14 (03) e1002267
    CrossrefPubMedGoogle Scholar
  • 9 Borchers RE, Gibson LJ, Burchardt H, Hayes WC. Effects of selected thermal variables on the mechanical properties of trabecular bone. Biomaterials 1995; 16 (07) 545-551
    CrossrefPubMedGoogle Scholar
  • 10 Smith CW, Young IS, Kearney JN. Mechanical properties of tendons: changes with sterilization and preservation. J Biomech Eng 1996; 118 (01) 56-61
    CrossrefPubMedGoogle Scholar
  • 11 Turner WD, Vasseur P, Gorek JE, Rodrigo JJ, Wedell JR. An in vitro study of the structural properties of deep-frozen versus freeze-dried, ethylene oxide-sterilized canine anterior cruciate ligament bone-ligament-bone preparations. Clin Orthop Relat Res 1988; (230) 251-256
    PubMedGoogle Scholar
  • 12 Foutz TL, Stone EA, Abrams Jr CF. Effects of freezing on mechanical properties of rat skin. Am J Vet Res 1992; 53 (05) 788-792
    PubMedGoogle Scholar
  • 13 Kiefer GN, Sundby K, McAllister D. , et al. The effect of cryopreservation on the biomechanical behavior of bovine articular cartilage. J Orthop Res 1989; 7 (04) 494-501
    CrossrefPubMedGoogle Scholar
  • 14 Callaghan JP, McGill SM. Frozen storage increases the ultimate compressive load of porcine vertebrae. J Orthop Res 1995; 13 (05) 809-812
    CrossrefPubMedGoogle Scholar
  • 15 Scott A, Wang X, Road JD, Reid WD. Increased injury and intramuscular collagen of the diaphragm in COPD: autopsy observations. Eur Respir J 2006; 27 (01) 51-59
    CrossrefPubMedGoogle Scholar
  • 16 Van Ee CA, Chasse AL, Myers BS. Quantifying skeletal muscle properties in cadaveric test specimens: effects of mechanical loading, postmortem time, and freezer storage. J Biomech Eng 2000; 122 (01) 9-14
    CrossrefPubMedGoogle Scholar
  • 17 Junqueira LCU, Bignolas G, Brentani RR. A simple and sensitive method for the quantitative estimation of collagen. Anal Biochem 1979; 94 (01) 96-99
    CrossrefPubMedGoogle Scholar
  • 18 Mayer RG, Reed JD. Embalming: history, theory, and practice. Norwalk, CT: Appleton & Lange; 1990
    Google Scholar
  • 19 Fitzgerald ER. Dynamic mechanical measurements during the life to death transition in animal tissues. Biorheology 1975; 12 (06) 397-408
    CrossrefPubMedGoogle Scholar
  • 20 Macgowan NA. Diaphragm injury in chronic respiratory disease. MSc thesis. Rehabilitation Sciences, University of British Columbia, Vancouver, Canada, 1998
    PubMedGoogle Scholar
  • 21 Rodrigues CJ, Rodrigues Junior AJ. A comparative study of aging of the elastic fiber system of the diaphragm and the rectus abdominis muscles in rats. Braz J Med Biol Res 2000; 33 (12) 1449-1454
    CrossrefPubMedGoogle Scholar
  • 22 Askanas V, Engel WK, Alvarez RB. Enhanced detection of congo-red-positive amyloid deposits in muscle fibers of inclusion body myositis and brain of Alzheimer's disease using fluorescence technique. Neurology 1993; 43 (06) 1265
    PubMedGoogle Scholar
  • 23 LaFerla FM, Green KN, Oddo S. Intracellular amyloid-β in Alzheimer's disease. Nat Rev Neurosci 2007; 8 (07) 499-509
    PubMedGoogle Scholar