ABSTRACT: Formalin has been used as a preservative for cadaver in anatomy for years now. 1st year MBBS students are not accustomed to this chemical earlier and have various reactions when they come in contact with this. Therefore we in the department of Anatomy, Bundelkhand Medical College, Sagar planned to study the effect of formalin on 150 1st year MBBS students by giving them questionnaire. In our study 30% had unpleasant smell of formalin,,24.67% students have reported irritation and watering of eyes, others reported symptoms includes running or congested nose (20.67%), Sore throat (17.33%), headache (14.67%), skin problems (12%), lack of concentration (16.67%) and many more. Finding an alternative for formalin in the form of phenoxyethanol is also a good option.

-> Effects of Formalin on First Year MBBS Students.pdf

ABSTRACT: Formalin is a widely used fixative but there is potential public health risks to exposure. Besides, alcoholic fixation is advantageous over formalin fixation because of faster fixation, optimal preservation and safer workplace environment. Following fixation by EMA and 10% neutral buffered formalin (NBF), we analyzed the tissue morphology, antigenic stability, DNA and RNA quantity with quality (OD value). The findings of EMA fixing on both the tissue morphology and molecular characterization, were satisfactory. Specially, EMA was faster in penetration of tissues than NBF, fixed ideally as early as 8 h of fixation whereas improper fixation was evident for NBF. In Hematoxylin and Eosin (H & E) staining, better cellular details with stronger affinity for staining were observed. In immunohistochemistry, better antigenic stability was reported for EMA-fixed tissues. The nucleic acid analysis revealed that total genomic DNA and RNA yield from EMA fixed tissues were significantly higher (P < 0.05) with superior quality than NBF fixed tissues. Our results suggest that EMA could be a potential alternative to NBF for fixation and preservation of tissues. These data provide new insights into an option for a safer working environment to support study and research.

-> Alcoholic fixation over formalin fixation.pdf

ABSTRACT: Formalin fixation and paraffin embedding (FFPE) is the standard method for tissue storage in histopathology. However, FFPE has disadvantages in terms of user health, environment, and nucleic acid integrity. Weigners fixative has been suggested as an alternative for embalming cadavers in human and veterinary anatomy. The present study tested the applicability of Weigners for histology and immunohistochemistry and the preservation of nucleic acids. To this end, a set of organs was fixed for 2 days and up to 6 months in Weigners (WFPE) or formalin. WFPE tissues from the skin, brain, lymphatic tissues, liver, and muscle had good morphologic preservation, comparable to formalin fixation. The quality of kidney and lung samples was inferior to FFPE material due to less accentuated nuclear staining and retention of proteinaceous interstitial fluids. Azan, Turnbull blue, toluidin, and immunohistochemical stainings for CD79a, cytokeratin, vimentin, and von Willebrand factor led to comparable results with both fixates. Of note, immunohistochemical detection of CD3 was possible after 6 months in WFPE but not in FFPE tissues. mRNA, miRNA, and DNA from WFPE tissues had superior quality and allowed for amplification of miRNA, 400-bp-long mRNA, and 1000-bp-long DNA fragments after 6 months of fixation in WFPE. In summary, Weigners fixative is a nonhazardous alternative to formalin, which provides a good morphologic preservation of most organs, a similar sensitivity for protein detection, and a superior preservation of nucleic acids. Weigners may therefore be a promising alternative to cryopreservation and may be embraced by people affected by formalin allergies.

-> Weigners Fixative.pdf

ABSTRACT: Formalin, widely utilized as a fixative in various fields including microscopy and histology, poses significant health risks due to its toxicity, often overlooked despite its prevalent use. This study aimed to assess the toxic effects of formalin on the health of first-year medical and dental students. Conducted over a year at the Department of Anatomy, SMBT Institute of Medical Sciences and Research Center, the cross-sectional study surveyed 200 students. Results from 180 respondents revealed a range of symptoms attributable to formalin exposure, including lacrimation, itching, running nose, redness, burning sensation in the eyes, and headaches. The prevalence of these symptoms underscores the importance of raising awareness among individuals working closely with formalin, such as embalmers and anatomists, as well as medical and dental students. Increased awareness can positively impact health outcomes, while further research may aid in mitigating occupational hazards and disseminating preventive measures and knowledge regarding formalin toxicity.

-> Effects of Formalin on Health of First Year Medical and Dental Students.pdf

ABSTRACT: Fixation is considered as physiochemical process where cells or tissues are fixed chem-ically. Fixatives perform various functions such as prevention of autolysis and tissue putrefaction. Various fixative agents include formaldehyde, glutaraldehyde, osmiumtetroxide, glyoxal, picric acid, and so on. A detailed search on PubMed, Google scholar, and Scopus database showed very few articles on “fixation” and “fixative.” Keeping this fact in mind, a comprehensive review on fixation and fixatives was prepared. The main aim of this review is to make pathologists and laboratory technicians familiar with the  basic aspects and different types of fixatives.

-> Fixation and Fixatives Roles and Functions—A Short.pdf

ABSTRACT: This study compared the effects of different fixatives on tissues for histopathology and molecular analysis. Forty tissues were fixed with six different fixatives, including formalin, Bouin, Hollande, Greenfix, UPM, and CyMol. Various staining and analysis techniques were applied. Results showed that alternative fixatives (Greenfix, UPM, CyMol) were equivalent to formalin in preserving cell morphology and antigenicity for immunohistochemistry. Hollande was the best for morphology, while Bouin was comparable to formalin. Alternative fixatives yielded suitable DNA for molecular analysis. Bouin-fixed tissues did not provide results comparable to formalin.

-> Application of alternative fixatives to formalin in diagnostic pathology.pdf

ABSTRACT: Formalin is a widely used fixative but there is potential public health risks to exposure. Besides, alcoholic fixation is advantageous over formalin fixation because of faster fixation, optimal preservation and safer workplace environment. Following fixation by EMA and 10% neutral buffered formalin (NBF), we analyzed the tissue morphology, antigenic stability, DNA and RNA quantity with quality (OD value). The findings of EMA fixing on both the tissue morphology and molecular characterization, were satisfactory. Specially, EMA was faster in penetration of tissues than NBF, fixed ideally as early as 8 h of fixation whereas improper fixation was evident for NBF. In Hematoxylin and Eosin (H & E) staining, better cellular details with stronger affinity for staining were observed. In immunohistochemistry, better antigenic stability was reported for EMA-fixed tissues. The nucleic acid analysis revealed that total genomic DNA and RNA yield from EMA fixed tissues were significantly higher (P < 0.05) with superior quality than NBF fixedtissues. Our results suggest that EMA could be a potential alternative to NBF for fixation and preservation of tissues. These data provide new insights into an option for a safer working environment to support study and research.

-> Saudi Journal of Biological Sciences.pdf

ABSTRACT: The study compares the efficacy of natural fixatives, specifically jaggery and Khandsari, against formalin, a widely used fixative in routine tissue processing. Ninety normal tissue specimens from minor oral surgical procedures were divided into three groups: 30% jaggery, 30% Khandsari, and 10% formalin. Evaluation of fixation quality, including cellular outline, cytoplasmic and nuclear details, staining quality, and overall morphology, revealed that Khandsari performed comparably to formalin, while jaggery showed somewhat inferior results. Specifically, Khandsari demonstrated excellent cellular outline, cytoplasmic staining, nuclear details, staining quality, and overall morphology, indicating its potential as an alternative fixative for human oral tissues. These findings suggest promising prospects for the use of Khandsari and jaggery as natural fixatives, offering safer alternatives to formalin in tissue fixation processes.

-> Natural substitutes for formalin A boon to histopathology!.pdf

ABSTRACT: Formalin is the principal tissue fxative used worldwide for clinical and research purposes. Despite optimal preservation of morphology, its preservation of DNA and RNA is poor. As clinical diagnostics increasingly incorporates molecular-based analysis, the requirement for maintaining nucleic acid quality is of increasing importance. Here we assess an alternative non-formalin-based tissue fxation method, PAXgene Tissue system, with the aim of better preserving nucleic acids, while maintaining the quality of the tissue to be used for vital existing diagnostic techniques. In this study, these criteria are assessed in a clinically representative setting. In total, 203 paired PAXgene Tissue and formalin-fxed samples were obtained. Blindscored haematoxylin and eosin (H&E) sections showed comparable and acceptable staining. Immunohistochemistry (IHC) staining was suboptimal using existing protocols but improved with minor method adjustment and optimisation. Quality of DNA and RNA was signifcantly improved by PAXgene tissue fxation [RIN 2.8 versus 3.8 (p<0.01), DIN 5.68 versus 6.77 (p<0.001)], which translated into improved performance on qPCR assay. These results demonstrate the potential of PAXgene Tissue to be used routinely in place of formalin, maintaining adequate histological staining and signifcantly improving the preservation of biological molecules in the genomic era.

-> Alternative tissue fxation for combined histopathological and molecular analysis in a clinically representative setting.pdf

ABSTRACT: In this study, three new alcohol-based fixatives, F-Solv, FineFIX, and RCL2, were evaluated alongside neutral buffered formalin (NBF) to address the toxicity and cross-linking issues associated with formaldehyde. Fresh tissue samples were divided and fixed in all fixatives for varying durations, followed by H&E staining, common histochemical stains, and immunohistochemical studies. While RCL2 fixation presented challenges due to tissue softness, F-Solv and FineFIX caused partial tissue disintegration during fixation. F-Solv exhibited morphological similarities to NBF but required substantial protocol adjustments for routine use. FineFIX induced shrinkage artifacts and cellular alterations, while RCL2 showed good morphological results but caused pigment deposition and cell damage. Despite positive attributes, none of the alcohol-based fixatives matched NBF in macroscopy, morphological evaluation, and immunohistochemical studies, emphasizing the continuing importance of NBF as the gold standard despite its environmental and safety concerns.

-> Formaldehyde Substitute Fixatives.pdf

ABSTRACT: Preservation of biological tissue is performed by using different fixative solutions. This fact had fundamental contribution on the history and scientific development of anatomy. The history of fixative solutions can be divided in three important moments: the first one is connected to the Old Egypt; the second, to the Renascence period; and the third period occurred during the American Civil War. Several solutions were tested and used throughout history and they adapted to the needs and contexts of each time. Currently, the main fixative solutions are toxic and some of them carcinogenic, including the formaldehyde, which remains as a “gold standard”. The fixative solutions are well adapted for histology laboratories, where their vapors are easily removed. On the other hand, in gross anatomy labs where usually not enough exhaustion exists, the exposure to toxic components is higher. Therefore, the necessity for better, more effective and safe solutions increased the interest for studying and several researchers have been engaged to develop a better fixative solution. This works brings a review of the historic progress of tissue fixation techniques for research and didactic purposes.

-> The use of fixative solutions throughout the ages a comprehensive review.pdf

ABSTRACT: Formaldehyde is commonly used worldwide, even though it is classified as carcinogenic to humans by the International Agency for Research on Cancer. This has motivated intensive investigations of formaldehyde substitutes, and recently, some alternative solutions were found, which can potentially replace it. Previous research showed that tannic acid (TA) in glutaraldehyde solution has the ability to stabilize elastin and collagen. This provided a basis for the development of a new alcoholic fixative solution, particularly aimed at extracellular matrix components, with TA as a main component. Heart, brain, and intestinal samples were fixed by immersion in 10% regular formalin solution (RFS), 70% ethanol solution (ES), and tannic acid ethanolic solution (TAES). Next, tissue fragments were prepared for routine histology procedures. The toxicity of TA was analyzed using in silico tests for mutagenicity, as well as for cutaneous and respiratory toxicity. Analyses of photomicrographs demonstrated that all fixative solutions have the ability to preserve the fragments.

The quantitative analyses showed that capability of TAES to preserve and stabilize elastin and collagen is superior to that of RFS and ES. We demonstrated that TA is not mutagenic, and it is less toxic for skin and respiratory tract. We therefore conclude that TAES can potentially represent a powerful and feasible alternative solution for fixing extracellular matrix of microscopic examination samples. Anat Rec, 301:1544–1550, 2018. © 2018 Wiley Periodicals, Inc.

-> Tannic Acid Solution A Better Fixative Solution Than Formalin for Elastin and Collagen—Toxic and Morphological.pdf

ABSTRACT: Formalin-fixed paraffin-embedded (FFPE) tissue is the predominant preparation for diagnostic histopathological evaluation and increasingly the biospecimen on which molecular diagnostics are performed. However, formalin is carcinogenic and results in cross-linking of proteins and nicking and alterations of nucleic acids. Alternative fixatives, including 70% ethanol, improved biomolecular integrity; however, they have yet to replace neutral-buffered formalin (NBF). Herein, we describe the phosphate-buffered ethanol 70% (BE70) fixative. The histomorphology of BE70-fixed tissue is very similar to that of NBF; however, it is a non-cross-linking fixative and lacks the carcinogenic profile of formaldehyde-based fixatives. RNA isolated from tissue fixed in BE70 was of substantially higher quality and quantity than that recovered from formalin-fixed tissue. Furthermore, the BE70 fixative showed excellent RNA and DNA integrity compared with that of NBF fixative based on real-time polymerase chain reaction analysis results. Immunohistochemical staining was similar for the antigen tested. In conclusion, BE70 is a non-cross-linking fixative that is superior to NBF and 70% ethanol with reference to biomolecule recovery and quality from paraffin-embedded tissue. Additional studies to compare the histomorphologic and immunohistochemical performance and utility in a clinical setting are required. (J Histochem Cytochem 64:425–440, 2016)

-> A Buffered Alcohol-Based Fixative for Histomorphologic and Molecular Applications.pdf