Fixation with formol gel*versus fixation with formalin Fixation with em Formol gel* versus fixation with formalin Study made at Centro Hospitalar de Setúbal, E.P.E- Portugal, by: Almas, S., Carretas, S., Fernandes, A., Silva, A. Silva, M.- Técnicas de Diagnóstico e terapêutica do Serviço de Anatomia Patológica do Centro Hospitalar de Setúbal, E.P.E. e Quintana, C. e Gonçalves M.- Anatomopatologistas do Serviço de Anatomia Patológica do Centro Hospitalar de Setúbal, E.P.E. * ANAPAT Formol gel, given by: Esteriplás - Produtos para a Saúde Lda. -Santa Maria da Feira Esteriplás- Dep. Técnico Page 1 Fixation with formol gel*versus fixation with formalin Introdution Fixation is the most important step during histological processing. Its purpose is to prevent autolysis and bacterial destruction of the tissues, maintaining its shape and volume during subsequent steps and keep the tissue closely as possible to its state in vivo. The fixative most used in routine of pathologic anatomy is 10% buffered formaldehyde as it is compatible with most complementary techniques that are used routinely in Pathology. Formaldehyde causes little shrinkage and distortion of tissue, has high penetration speed, since diffuses rapidly within the tissues, however it possesses a low fixing speed, slow to denature and precipitate proteins tissue, this being the form of stabilization the chemical components of tissues. Formaldehyde in liquid form has the disadvantage of producing almost immediately, vapors irritating to the conjunctiva, upper respiratory tract and skin. However the same fixative product is available at gel form, whose advantage is the reduction in the formation of irritating vapors, allowing the operator to be exposed during longer periods without feeling the effects mentioned above. Because of its consistency, prevents accidental splashing and spillage during transport and handling of biological samples. Objetives: The objective of this study is to compare the morphology, quality and preservation of the biological samples fixed in 10% buffered formaldehyde in liquid and gel form proceeding to macroscopic and microscopic evaluations: - The macroscopic evaluation, compare the visible effects in tissues exposed to formaldehyde buffered 10% liquid and gel. - In microscopy evaluation, through the use of routine staining and complementary diagnostic techniques (histochemistry and immunocytochemistry), comparing the morphology, consistent quality and preservation of the tissues subject to two kinds of fasteners. Methodology: -Samples: Uterus, Kidney, Colon and Gall, welcomed in fresh and sectioned into equal parts for both fasteners. -Fixation: Liquid formaldehyde and formaldehyde gel * - Fixation time: 8h, 24h, 48h, 72h Esteriplás- Dep. Técnico Page 2 Fixation with formol gel*versus fixation with formalin Evaluation Criteria: Macroscopy: Method observational Microscopy: - H & E staining for morphological and preservation. - Imunocitoquimica-AE1/AE3 (Colon), Vim (kidney), alpha-actin (uterus), CD34 (gall) to evaluate the feasibility of etípodos. - Histochemistry Masson, to assess the viability of the tissue components. Results: Tissue type Fixation time Fixator kind Macroscopie Morfology H&E Preservation Immunocytochemistry Histochemistry QUALItY uterus kidney gall COLÓN 24H 48H 72H 24H 48H 72H 8H 24H 48H 24H 48H 72H L G L G L G L G L G L G L G L G L G L G L G L G 2 2 4 4 4 4 2 2 4 4 4 4 2 2 4 4 4 4 2 2 4 4 4 4 4 4 4 4 3 3 4 4 4 4 4 4 2 2 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 2 2 3 3 4 4 4 4 4 4 4 4 3 3 4 3 4 2 2 2 3 3 4 4 2 2 3 3 4 4 3 3 3 3 3 3 3 4 3 3 3 4 4 4 4 4 3 4 4 4 4 4 4 4 3 3 3 3 4 4 16 17 19 18 18 17 16 16 19 19 19 20 12 12 17 17 20 20 16 16 18 18 19 19 Legend: L = liquid G = Gel Classification: 0=Absence; 1= weak ; 2=Moderate ; 3= Strong ; 4= Very strong Quality: It results from the sum of the evaluation criteria Esteriplás- Dep. Técnico Page 3 Fixation with formol gel*versus fixation with formalin Conclusion: During handling of the samples, it was found that by placing the sample container in the gel, it is immediately immersed in contrast to what happens sometimes in formalin liquid, where samples float. There was also a decrease of accidental splashing as well as the decrease in strong odors released by formaldehyde liquid. Regarding the evaluation criteria, it was found that the results are superimposable on either macroscopically or in morphology and tissue preservation; slight variations occurred but they were not significant and did not hamper the use of complementary techniques. Regarding the different time of fixation the differences aren’t significant exception for 8h clamping used in gall, whose results were worse. We decided not to use fixture time for the remaining samples of the study. Despite the small sample size of this study, we found that the formaldehyde gel is feasible for routine laboratory in Pathology, which is consistent with the study conducted by Michael LaFrinieri. Esteriplás- Dep. Técnico Page 4