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Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*

Year 2020, Volume: 17 Issue: 3, 242 - 247, 01.12.2020
https://doi.org/10.32707/ercivet.828418

Abstract

Tespit işlemi, hücre ve doku komponentlerinin ölüm sonrasında morfolojik özelliklerinin canlıdaki durumuna en yakın şekilde sabitleyip, histolojik olarak incelenmesine olanak tanır. Tespit işleminde kullanılan kimyasal ajanlara fik-satif denir. Yirminci yüzyıldan günümüze kadar rutin doku tespit işleminde formaldehit çözeltisi kullanılmaktadır. Her fiksatif doku tespit işlemi süresince doku histomorfolojisinin incelenmesi adına avantaj ve dezavantajlara sahiptir. Bu nedenle tespit işlemi için amaca yönelik doğru fiksatif ile çalışılması gerekir. Bu çalışmada hedef, 3 farklı fiksatif ile hazırlanan 4 çözeltinin, diğer faktörler optimum seviyede tutularak, doku tespit işlemi sürecinde nasıl etkilere sahip olduklarını belirlemek ve tespit özelliklerine göre en ideal kullanım amaçlarını ortaya çıkarmaktı. Bu amaçla, çalışmada doku tespit işlemi sırasında fiksatiflerin dezavantajlarından etkilenen ve hücre morfolojisinin incelenmesinde zorluklarla karşılaşılan dalak ve böbrek dokuları üzerinde çalışıldı. Yaklaşık 0.5 cm kalınlığındaki doku örnekleri hazırlanan %3.7`lik, %10`luk formaldehit ve Hollande çözeltilerinde 16 saat, Carnoy çözeltisinde ise 4 saat süre ile tespit edildi. Ru-tin doku takip işlemlerinden geçirildikten sonra alınan doku kesitleri H&E ile boyandı. Bu dokuların histomorfolojik ince-lemeleri yapıldığında hücre sitoplazma boyanması için en uygun fiksatifin %3.7`lik formaldehit çözeltisi olduğu, hücre çekirdek ve çekirdekçiklerinin morfolojisini ortaya koymada ise Carnoy ve Hollande çözeltilerin kullanımlarının ideal olduğu, ancak Carnoy çözeltisinin eritrositlerde ileri düzeyde lizise yol açtığı gözlendi. Sonuç olarak, doku tespit işle-minde %3.7`lik formaldehit çözeltisinin kullanımı ideal bulunurken, Carnoy ve Hollande çözeltilerinin ise hücre çekirdek-lerindeki histomorfolojik değişikliklerin detaylı incelenmesini gerektiren hastalıklarda etkili bir kullanım oluşturacakları ön görüldü.

References

  • Ahmed HG, Mohammed AI, Hussein MOM. A com-parison study of histochemical staining of various tissues after Carnoy’s and formalin fixation. Sud J Med Sc 2010; 5(4): 277-83.
  • Baker FJ, Silverton RE. Introduction to Medical La-boratory Technology. Fifth Edition. London: Butterworth & Co Ltd. 1976; p. 312.
  • Bancroft JD, Gamble M. Theory and Practice of His-tological Techniques. Sixth Edition. Philadelphia, PA: Churchill Livingstone/Elsevier, 2008; p. 53-75.
  • Benerini Gatta L, Cadei M, Balzarini P, Castriciano S, Paroni R, Verzeletti A, Cortellini V, De Ferrari F, Grigolato P. Application of alternative fixatives to formalin in diagnostic pathology. Eur J Histochem 2012; 56(2): e12.
  • Coleman WB, Tsongalis GJ. Molecular Diagnostics: For the Clinical Laboratorian. Second Edition. New Jersey: Humana Press, Clifton, 2006; p. 204.
  • Crawford NC, Barer R. The action of formaldehyde on living cells as studied by phase-contrast mi-croscopy. Q J Microsc Sci 1951; 3(20): 403-452.
  • Doğan, Ö. Histopatolojik tanı sürecinde standard-izasyon. APJ 2005; 2: 8-28.
  • Drury RAB, Wallington EA. Carleton Histological Techniques. Fifth Edition. London: Oxford Univer-sity Press, 1980; p. 36-44.
  • Eltoum I, Fredenburgh J, Myers RB, Grizzle WE. Introduction to the theory and practice of fixation of tissues. J Histotechnol 2001; 24(3): 173-90.
  • Fox CH, Johnson FB, Whiting J, Roller PP. Formal-dehyde fixation. J Histochem Cytochem 1985; 33(8): 845-53.
  • Freeman LEB, Blair A, Lubin JH, Stewart PA, Hayes RB, Hoover RN, Hauptmann M. Mortality from lymphohematopoietic malignancies among work-ers in formaldehyde industries: The national can-cer institute cohort. J Natl Cancer Inst 2009; 101(10): 751-61.
  • Gillespie JW, Best CJ, Bichsel VE, Cole KA, Green-hut SF, Hewitt SM, Ahram M, Gathright YB, Meri-no MJ, Strausberg RL, Epstein JI, Hamilton SR, Gannot G, Baibakova GV, Calvert VS, Flaig MJ, Chuaqui RF, Herring JC, Pfeifer J, Petricoin EF, Linehan WM, Duray PH, Bova GS, Emmert-Buck MR. Evaluation of non-formalin tissue fixation for molecular profiling studies. Am J Pathol 2002; 160(2):449-57.
  • Grizzle WE. The use of fixatives in diagnostic pathol-ogy. J Histotechnol 2001; 24(3): 151-2.
  • Grizzle WE. Models of fixation and tissue processing. Biotech Histochem 2009; 84(5): 185-93.
  • Kahyaoğlu F, Gökçimen A. Light microscopic deter-mination of tissue. East J Med 2017; 22(3): 120-4.
  • Luz DABP, Ribeiro U Jr, Chassot C, Collet E, De Salles Collet e Silva F, Cecconello I, Corbett CE. Camoy’s solution enchances lymph node detec-tion: An anatomical dissection study in cadavers. Histopathology 2008; 53(6): 740-2.
  • Pereira MA, Dias AR, Faraj SF, Cirqueira Cdos S, Tomitao MT, Nahas SC, Ribeiro U Jr, de Mello ES. Carnoy's solution is an adequate tissue fixa-tive for routine surgical pathology, preserving cell morphology and molecular integrity. Histopathology 2015; 66(3): 388-97.
  • Resch A, Langner C. Lymph node staging in colorec-tal cancer: Old controversies and recent advanc-es. World J Gastroenterol 2013; 19(46): 8515-26.
  • Singhal P, Singh NN, Sreedhar G, Banerjee S, Batra M, Garg A. Evaluation of histomorphometric changes in tissue architecture in relation to altera-tion in fixation protocol - an invitro study. J Clin Diagn Res 2016; 10(8): 28-32.
  • Warmington AR, Wilkinson JM, Riley CB. Evaluation of ethanol-based fixatives as a substitute for for-malin in diagnostic clinical laboratories. J Histo-technol 2000; 23(4): 299-308.

Fixation and Comparative Histologic Evaluation of Cat and Dog Tissues using Different Fixatives

Year 2020, Volume: 17 Issue: 3, 242 - 247, 01.12.2020
https://doi.org/10.32707/ercivet.828418

Abstract

Fixation is a procedure that allows histological evaluation of cells and tissue components by fixing them to their closest morphological features in life-like state after death. Formaldehyde solution has been in use since the 20th century. During the tissue fixation, each fixatives represent their own advantages and disadvantages on the examina-tion of histomorphological properties of the tissues. So that, the fixation has to be performed with the appropriate fixa-tive depending on the research aim. The aim of this study was to determine the effects of four solutions, prepared by 3 different fixatives, on the tissue fixation process by keeping the other factors at an optimum level and to determine the most ideal use according to the fixation properties. For this purpose, the spleen and kidney tissues were studied, since they are generally affected by the disadvantages of fixatives during the tissue fixation process and lead to difficulties in examining cell morphology. Tissue samples of 0.5 cm thickness were fixed in the 3.7%, 10% formaldehyde and Hol-lande solutions for 16 hours and Carnoy solution for 4 hours. Thereafter, tissue samples were routinely proceeded, and sections were stained by H&E. The histomorphological examinations of these tissues revealed that the most suitable fixative for cytoplasmic staining was 3.7% formaldehyde solution, while the use of Carnoy and Hollande solutions was ideal in revealing the morphology of the nucleus and nucleolus. However the Carnoy solution caused high lysis in erythrocytes. As a result, it was concluded that the 3.7% formaldehyde solution could be ideally used in tissue fixation process, whereas Carnoy and Hollande solutions were seen to be effective in diseases that require detailed examina-tion of histomorphological changes in nucleus and nucleolus.

References

  • Ahmed HG, Mohammed AI, Hussein MOM. A com-parison study of histochemical staining of various tissues after Carnoy’s and formalin fixation. Sud J Med Sc 2010; 5(4): 277-83.
  • Baker FJ, Silverton RE. Introduction to Medical La-boratory Technology. Fifth Edition. London: Butterworth & Co Ltd. 1976; p. 312.
  • Bancroft JD, Gamble M. Theory and Practice of His-tological Techniques. Sixth Edition. Philadelphia, PA: Churchill Livingstone/Elsevier, 2008; p. 53-75.
  • Benerini Gatta L, Cadei M, Balzarini P, Castriciano S, Paroni R, Verzeletti A, Cortellini V, De Ferrari F, Grigolato P. Application of alternative fixatives to formalin in diagnostic pathology. Eur J Histochem 2012; 56(2): e12.
  • Coleman WB, Tsongalis GJ. Molecular Diagnostics: For the Clinical Laboratorian. Second Edition. New Jersey: Humana Press, Clifton, 2006; p. 204.
  • Crawford NC, Barer R. The action of formaldehyde on living cells as studied by phase-contrast mi-croscopy. Q J Microsc Sci 1951; 3(20): 403-452.
  • Doğan, Ö. Histopatolojik tanı sürecinde standard-izasyon. APJ 2005; 2: 8-28.
  • Drury RAB, Wallington EA. Carleton Histological Techniques. Fifth Edition. London: Oxford Univer-sity Press, 1980; p. 36-44.
  • Eltoum I, Fredenburgh J, Myers RB, Grizzle WE. Introduction to the theory and practice of fixation of tissues. J Histotechnol 2001; 24(3): 173-90.
  • Fox CH, Johnson FB, Whiting J, Roller PP. Formal-dehyde fixation. J Histochem Cytochem 1985; 33(8): 845-53.
  • Freeman LEB, Blair A, Lubin JH, Stewart PA, Hayes RB, Hoover RN, Hauptmann M. Mortality from lymphohematopoietic malignancies among work-ers in formaldehyde industries: The national can-cer institute cohort. J Natl Cancer Inst 2009; 101(10): 751-61.
  • Gillespie JW, Best CJ, Bichsel VE, Cole KA, Green-hut SF, Hewitt SM, Ahram M, Gathright YB, Meri-no MJ, Strausberg RL, Epstein JI, Hamilton SR, Gannot G, Baibakova GV, Calvert VS, Flaig MJ, Chuaqui RF, Herring JC, Pfeifer J, Petricoin EF, Linehan WM, Duray PH, Bova GS, Emmert-Buck MR. Evaluation of non-formalin tissue fixation for molecular profiling studies. Am J Pathol 2002; 160(2):449-57.
  • Grizzle WE. The use of fixatives in diagnostic pathol-ogy. J Histotechnol 2001; 24(3): 151-2.
  • Grizzle WE. Models of fixation and tissue processing. Biotech Histochem 2009; 84(5): 185-93.
  • Kahyaoğlu F, Gökçimen A. Light microscopic deter-mination of tissue. East J Med 2017; 22(3): 120-4.
  • Luz DABP, Ribeiro U Jr, Chassot C, Collet E, De Salles Collet e Silva F, Cecconello I, Corbett CE. Camoy’s solution enchances lymph node detec-tion: An anatomical dissection study in cadavers. Histopathology 2008; 53(6): 740-2.
  • Pereira MA, Dias AR, Faraj SF, Cirqueira Cdos S, Tomitao MT, Nahas SC, Ribeiro U Jr, de Mello ES. Carnoy's solution is an adequate tissue fixa-tive for routine surgical pathology, preserving cell morphology and molecular integrity. Histopathology 2015; 66(3): 388-97.
  • Resch A, Langner C. Lymph node staging in colorec-tal cancer: Old controversies and recent advanc-es. World J Gastroenterol 2013; 19(46): 8515-26.
  • Singhal P, Singh NN, Sreedhar G, Banerjee S, Batra M, Garg A. Evaluation of histomorphometric changes in tissue architecture in relation to altera-tion in fixation protocol - an invitro study. J Clin Diagn Res 2016; 10(8): 28-32.
  • Warmington AR, Wilkinson JM, Riley CB. Evaluation of ethanol-based fixatives as a substitute for for-malin in diagnostic clinical laboratories. J Histo-technol 2000; 23(4): 299-308.
There are 20 citations in total.

Details

Primary Language Turkish
Journal Section Articles
Authors

Hazal Öztürk Gürgen This is me 0000-0003-2748-6189

Funda Yıldırım This is me

Publication Date December 1, 2020
Submission Date February 4, 2020
Acceptance Date August 17, 2020
Published in Issue Year 2020 Volume: 17 Issue: 3

Cite

APA Öztürk Gürgen, H., & Yıldırım, F. (2020). Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, 17(3), 242-247. https://doi.org/10.32707/ercivet.828418
AMA Öztürk Gürgen H, Yıldırım F. Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*. Erciyes Üniv Vet Fak Derg. December 2020;17(3):242-247. doi:10.32707/ercivet.828418
Chicago Öztürk Gürgen, Hazal, and Funda Yıldırım. “Kedi Ve Köpek Dokularının Farklı Fiksatiflerle Tespiti Ve Histolojik Görünümlerinin Karşılaştırılması*”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17, no. 3 (December 2020): 242-47. https://doi.org/10.32707/ercivet.828418.
EndNote Öztürk Gürgen H, Yıldırım F (December 1, 2020) Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17 3 242–247.
IEEE H. Öztürk Gürgen and F. Yıldırım, “Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*”, Erciyes Üniv Vet Fak Derg, vol. 17, no. 3, pp. 242–247, 2020, doi: 10.32707/ercivet.828418.
ISNAD Öztürk Gürgen, Hazal - Yıldırım, Funda. “Kedi Ve Köpek Dokularının Farklı Fiksatiflerle Tespiti Ve Histolojik Görünümlerinin Karşılaştırılması*”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi 17/3 (December 2020), 242-247. https://doi.org/10.32707/ercivet.828418.
JAMA Öztürk Gürgen H, Yıldırım F. Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*. Erciyes Üniv Vet Fak Derg. 2020;17:242–247.
MLA Öztürk Gürgen, Hazal and Funda Yıldırım. “Kedi Ve Köpek Dokularının Farklı Fiksatiflerle Tespiti Ve Histolojik Görünümlerinin Karşılaştırılması*”. Erciyes Üniversitesi Veteriner Fakültesi Dergisi, vol. 17, no. 3, 2020, pp. 242-7, doi:10.32707/ercivet.828418.
Vancouver Öztürk Gürgen H, Yıldırım F. Kedi ve Köpek Dokularının Farklı Fiksatiflerle Tespiti ve Histolojik Görünümlerinin Karşılaştırılması*. Erciyes Üniv Vet Fak Derg. 2020;17(3):242-7.