MyMedR

Natural fixatives alternative to formalin in histopathology: A systematic review

Yee AWM, Oo PS, Aye SN, Lim WJ, Chee VCX, Krishnappa P

Med J Malaysia, 2023 Jan;78(1):98-108.

INTRODUCTION: Since constant long-term exposure to formaldehyde endangers the health of laboratory personnel, sugar-based natural products have become interesting alternative fixatives to formaldehyde because of their preservative and antibacterial properties. However, there are controversial findings on the fixative effects of natural fixatives. This study systematically reviews the evidence comparing natural fixatives' types, dilutions, fixative properties and staining quality in normal tissues and histopathological specimens.

MATERIALS AND METHODS: A comprehensive search was performed for studies comparing the natural fixatives- and formaldehyde-fixed tissues using databases from inception to January 2022: PubMed, Ovid Medline and Google Scholar. Two independent reviewers did data extraction. The data were pooled for the type of natural fixatives, their concentrations and fixative qualities compared to formaldehyde.

RESULTS: Fifteen studies were included in this systematic review. Nine studies used one natural fixative with different dilutions, while six used several natural fixatives to compare their fixative properties with formaldehyde. The most used natural fixative was honey (n = 12) followed by jaggery (n = 8), sugar (n = 3) and others (n = 1). Honey showed the most promising results in fixation and staining, which are compatible with formalin. Jaggery and sugar also showed the possibility of replacing formaldehyde in tissue fixation and staining in smaller tissue samples.

CONCLUSION: Natural fixatives showed promising results in tissue fixation. However, optimising the concentrations and conditions of natural fixatives is difficult because of the different chemical constituents and production steps. More comprehensive studies are necessary for application.

Matched MeSH terms: Fixatives/pharmacology; Fixatives/chemistry

Microwave-stimulated fixation for electron microscopy using a domestic microwave oven

Ng KH, Gan SK

Malays J Pathol, 1990 Jun;12(1):27-33.

PMID: 2090887

We investigated microwave-stimulated fixation of tissues for transmission electron microscopy using a domestic microwave oven operating at a frequency of 2.45 GHz with an output power of 500W. Microwave-stimulated fixation, in 4% glutaraldehyde, of fresh rat kidney, liver, heart and brain tissues was compared to conventional fixation. Human renal biopsies were similarly studied. Electron microscopy showed excellent ultrastructural preservation comparable to that obtained by conventional fixation. The optimal temperature range for microwave-stimulated fixation was found to lie between 50 degrees C and 55 degrees C. Our results indicate that microwave-stimulated fixation is a rapid and reproducible technique and can be effectively applied to routine diagnostic pathology.

Matched MeSH terms: Fixatives*

Prevention of mother-to-child transmission of hepatitis B virus: An observation of routine practice in a tertiary liver centre before and after the introduction of the global health sector strategy on viral hepatitis

Hoo CZ, Wan Abdullah WZ, Omar H, Tan SS

Med J Malaysia, 2023 Mar;78(2):234-240.

PMID: 36988536

INTRODUCTION: Worldwide, around 296 million people have hepatitis B virus (HBV) infection, most commonly transmitted from mother-to-child. Global Health Sector Strategy on Viral Hepatitis (GHSSVH) was introduced in May 2016, calling for elimination of viral hepatitis by 2030. This study aims to compare practice in a tertiary liver centre before and after GHSSVH introduction for prevention of mother-to-child transmission (MTCT).

MATERIALS AND METHODS: This retrospective cohort study was performed in a tertiary referral liver centre in Malaysia, using data from electronic medical record from January 2015 to December 2019. A total of 1457 medical records of female with HBV infection were screened. The inclusion criteria of the study were pregnant women with HBsAg positive or known to have HBV infection during the study period. We excluded patients with co-infections of other types of viral hepatitis or human immunodeficiency virus, concurrent liver diseases (e.g.: autoimmune hepatitis, Wilson’s disease), previous organ transplant and malignancy—except for hepatocellular carcinoma (HCC).

RESULTS: This study included 117 pregnancies and 21/117 (17.9%) were on antiviral therapy (AVT) for HBV. In 2017– 2019, 13/18 (72.2%) of those with HBV DNA >200,000IU/ml were on AVT, compared to 5/9 (55.6%) for 2015–2016, indicating 58% (95% CI −63% to 568%) higher odds of being on AVT in post GHSSVH group after accounting for HBV DNA.

CONCLUSION: Uptake of maternal AVT for the prevention of MTCT shows an increased trend since the introduction of GHSSVH, with room for improvement.

Matched MeSH terms: Fixatives/pharmacology

Fulltext Optimization of the cytogenetic protocol for Pangasianodon hypophthalmus (Sauvage, 1878) and Clarias gariepinus (Burchell, 1822)

Okomoda VT, Koh ICC, Hassan A, Amornsakun T, Moh JHZ, Shahreza SM

PeerJ, 2018;6:e5712.

PMID: 30416879 DOI: 10.7717/peerj.5712

To obtain well spread chromosomes, the cytogenetic protocol for Pangasianodon hypophthalmus and Clarias gariepinus were optimized. This includes, the colchicine concentration (0.01%, 0.025%, 0.05%)/exposure duration (1, 3, and 5 h), hypotonic solution (distilled water or 0.075M KCl solution)/exposure duration (30 min, 1, and 2 h), the time of cell suspension preparation (at hypotonic treatment or before slide preparation) and chromosome aging period (0, 3, and 7 days in Carnoy's fixative). In addition, the type (i.e., fin, gill or kidney) and the amount of tissue (10, 50, 100 or 150 mg) were also investigated. Regardless of the species, the result obtained showed that well-spread chromosomes could be obtained using the following optimized protocol: Juveniles are injected with 0.05% colchicine (at one ml kg-1) and allowed to swim for 3 h. Then, 50 mg of gill tissue is made into cell suspension in 0.075M KCl for 1 h. The cell suspension is treated in Carnoy's fixative (changed three times at 20 min interval) and then aged for 3 days. Finally, chromosome slides are made and stained with 10% Giemsa for 1 h.

Matched MeSH terms: Fixatives

Glutaraldehyde in dentistry--a review

Rusmah M

Singapore Dent J, 1993 Jun;18(1):17-21.

PMID: 9582689

The disinfective and fixative properties of glutaraldehyde are now widely investigated. Glutaraldehyde is effective against micro-organisms and their spores. Recently, studies have shown the effectiveness of glutaraldehyde against the HIV virus. 2% glutaraldehyde is now recommended for the sterilisation of surgical instruments, operating areas, dental impressions and root canals during endodontic therapy. Studies have also shown that glutaraldehyde is an effective fixative with minimum side effects, limited penetration and quick acting. Pulpotomy studies using glutaraldehyde as the fixative agent produce high success rates. The important feature is the vital pulpal tissue at the apical third suggesting its limited penetration. The small amounts that get distributed systemically are quickly metabolised and excreted in the urine or exhaled as carbon dioxide.

Matched MeSH terms: Fixatives/pharmacokinetics; Fixatives/pharmacology*

Fulltext The Adverse Effect Of Formalin: A Warning Against Mishandling

Lian, C.B., Ngeow, W.C.

Ann Dent, 2000;7(1):-.

MyJurnal

Formalin is a clear solution of 37% formaldehyde in water. It is used in dentistry as a disinfectant, antiseptic and mainly as tissue fixative for preserving biologic specimens for histopathologic examination. The human knowledge on systemic formaldehyde intoxication is inadequate as only few cases of formalin ingestion have been reported. This paper presents a brief communication of the adverse effect of formalin to the human tissue.

Matched MeSH terms: Fixatives

RCL2, a potential formalin substitute for tissue fixation in routine pathological specimens

Masir N, Ghoddoosi M, Mansor S, Abdul-Rahman F, Florence CS, Mohamed-Ismail NA, et al.

Histopathology, 2012 Apr;60(5):804-15.

PMID: 22320393 DOI: 10.1111/j.1365-2559.2011.04127.x

To investigate RCL2 as a fixative for tissue fixation in routine histopathological examination and to assess tissue suitability for ancillary investigations.

Matched MeSH terms: Fixatives*

Fulltext In vivo analgesic effect of aqueous extract of Tamarindus indica L. fruits

Khalid S, Shaik Mossadeq WM, Israf DA, Hashim P, Rejab S, Shaberi AM, et al.

Med Princ Pract, 2010;19(4):255-9.

PMID: 20516700 DOI: 10.1159/000312710

To study the effects of Tamarindus indica L. aqueous fruit extract on the antinociceptive activities in rodent models.

Matched MeSH terms: Fixatives/adverse effects

Specimen shrinkage and its influence on margin assessment in breast cancer

Yeap BH, Muniandy S, Lee SK, Sabaratnam S, Singh M

Asian J Surg, 2007 Jul;30(3):183-7.

PMID: 17638637

The determination of tumour-free margin in breast cancer is crucial in deciding subsequent patient management. To exemplify the phenomenon of margin contraction during specimen preparation for histopathological analysis, we quantified the shrinkage of breast specimens as a result of formalin fixation.

Matched MeSH terms: Fixatives/pharmacology

Mosquito coil emissions and health implications

Liu W, Zhang J, Hashim JH, Jalaludin J, Hashim Z, Goldstein BD

Environ Health Perspect, 2003 Sep;111(12):1454-60.

PMID: 12948883

Burning mosquito coils indoors generates smoke that can control mosquitoes effectively. This practice is currently used in numerous households in Asia, Africa, and South America. However, the smoke may contain pollutants of health concern. We conducted the present study to characterize the emissions from four common brands of mosquito coils from China and two common brands from Malaysia. We used mass balance equations to determine emission rates of fine particles (particulate matter < 2.5 microm in diameter; PM(2.5)), polycyclic aromatic hydrocarbons (PAHs), aldehydes, and ketones. Having applied these measured emission rates to predict indoor concentrations under realistic room conditions, we found that pollutant concentrations resulting from burning mosquito coils could substantially exceed health-based air quality standards or guidelines. Under the same combustion conditions, the tested Malaysian mosquito coils generated more measured pollutants than did the tested Chinese mosquito coils. We also identified a large suite of volatile organic compounds, including carcinogens and suspected carcinogens, in the coil smoke. In a set of experiments conducted in a room, we examined the size distribution of particulate matter contained in the coil smoke and found that the particles were ultrafine and fine. The findings from the present study suggest that exposure to the smoke of mosquito coils similar to the tested ones can pose significant acute and chronic health risks. For example, burning one mosquito coil would release the same amount of PM(2.5) mass as burning 75-137 cigarettes. The emission of formaldehyde from burning one coil can be as high as that released from burning 51 cigarettes.

Matched MeSH terms: Fixatives/analysis

Intracellular antigen determination by flow cytometry in leukaemic cells

Chin SF, Cheong SK

Malays J Pathol, 1994 Jun;16(1):69-73.

PMID: 16329579

Several fixation and permeabilization techniques that enable the flow cytometric analysis of the cell contents have been introduced in recent years. These methods allow sensitive detection of intracellular antigens that facilitates the diagnosis of certain diseases. We have undertaken in this study to evaluate a simple method of fixation and permeabilization using 2% paraformaldehyde and Tween 20. Intracellular antigens in three different leukaemia cases were analysed. We found that the method was reliable and easy. Intracellular kappa light chains were found in abundance in a case of plasma cell leukaemia. CD3 and CD22 were found in greater amount intracellularly than on the surface in pre-T-ALL and pre-pre B-ALL respectively.

Matched MeSH terms: Fixatives

Detection of Candida albicans ADH1 and ADH2 mRNAs in human archival oral biopsy samples

Bakri MM, Cannon RD, Holmes AR, Rich AM

J Oral Pathol Med, 2014 Oct;43(9):704-10.

PMID: 24931506 DOI: 10.1111/jop.12193

The aim of this study was to investigate the relationship between expression of Candida albicans alcohol dehydrogenases (ADH) genes in archival formalin-fixed paraffin-embedded (FFPE) samples from biopsies of leukoplakia.

Matched MeSH terms: Fixatives

Fulltext Molecular markers associated with nonepithelial ovarian cancer in formalin-fixed, paraffin-embedded specimens by genome wide expression profiling

Vui-Kee K, Mohd Dali AZ, Mohamed Rose I, Ghazali R, Jamal R, Mokhtar NM

Kaohsiung J. Med. Sci., 2012 May;28(5):243-50.

PMID: 22531302 DOI: 10.1016/j.kjms.2011.11.007

Nonepithelial ovarian cancer (NEOC) is a rare cancer that is often misdiagnosed as other malignant tumors. Research on this cancer using fresh tissues is nearly impossible because of its limited number of samples within a limited time provided. The study is to identify potential genes and their molecular pathways related to NEOC using formalin-fixed paraffin embedded samples. Total RNA was extracted from eight archived NEOCs and seven normal ovaries. The RNA samples with RNA integrity number >2.0, purity >1.7 and cycle count value <28 cycles were hybridized to the Illumina Whole-Genome DASL assay (cDNA-mediated annealing, selection, extension, and ligation). We analyzed the results using the GeneSpring GX11.0 and FlexArray software to determine the differentially expressed genes. Microarray results were validated using an immunohistochemistry method. Statistical analysis identified 804 differentially expressed genes with 443 and 361 genes as overexpressed and underexpressed in cancer, respectively. Consistent findings were documented for the overexpression of eukaryotic translation elongation factor 1 alpha 1, E2F transcription factor 2, and fibroblast growth factor receptor 3, except for the down-regulated gene, early growth response 1 (EGR1). The immunopositivity staining for EGR1 was found in the majority of cancer tissues. This finding suggested that the mRNA level of a transcript did not always match with the protein expression in tissues. The current gene profile can be the platform for further exploration of the molecular mechanism of NEOC.

Matched MeSH terms: Fixatives

Displaying all 13 publications