Novel tissue preservative and tissue fixative for comparative pathology and animal research

Gugic, Dijana; Nassiri, Mehdi; Nadji, Mehrdad; Morales, Azorides R.; Vincek, Vladimir

doi:10.1016/j.jeas.2006.09.002

Cited by 14 publications

(19 citation statements)

References 27 publications

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“…Antibody specifically used in immuno-peroxidase staining method; needs to be studied in fresh tissue, which gives no result in formaldehyde fixed tissues. This is because the antigenic epitope by the formaldehyde solution is modified or destroyed (9). Formaldehyde cross-links molecular groups to prevent immunohistochemical reactions (6).…”

Section: Introductionmentioning

confidence: 99%

Light microscopic determination of tissue

Kahyaoğlu¹,

Gökçimen²

2017

Eastern J Med

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Section: Introductionmentioning

confidence: 99%

Light microscopic determination of tissue

Kahyaoğlu¹,

Gökçimen²

2017

Eastern J Med

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“…Unfortunately, RNA later alone resulted in uneven immunohistochemical staining and preserved noticeably less of the finer structural details compared to formalin (Paska et al, 2004). This could be improved by post-fixing with formalin, but formalin is known to damage RNA (Gugic et al, 2007; Masuda et al, 1999). …”

Section: Introductionmentioning

confidence: 99%

Molecular fixative enables expression microarray analysis of microdissected clinical cervical specimens

Li¹,

Niekerk

Miller

et al. 2014

Experimental and Molecular Pathology

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Formalin-fixed tissue has been a mainstay of clinical pathology laboratories, but formalin alters many biomolecules, including nucleic acids and proteins. Meanwhile, frozen tissues contain better-preserved biomolecules, but tissue morphology is affected, limiting their diagnostic utility. Molecular fixatives promise to bridge this gap by simultaneously preserving morphology and biomolecules, enabling clinical diagnosis and molecular analyses on the same specimen. While previous reports have broadly evaluated the use of molecular fixative in various human tissues, we present here the first detailed assessment of the applicability of molecular fixative to both routine histopathological diagnosis and molecular analysis of cervical tissues. Ten specimens excised via the Loop Electrosurgical Excision Procedure, which removes conical tissue samples from the cervix, were cut into alternating pieces preserved in either formalin or molecular fixative. Cervical specimens preserved in molecular fixative were easily interpretable, despite featuring more eosinophilic cytoplasm and more recognizable chromatin texture than formalin-fixed specimens. Immunohistochemical staining patterns of p16 and Ki-67 were similar between fixatives, although Ki-67 staining was stronger in the molecular fixative specimens. The RNA of molecular fixative specimens from seven cases representing various dysplasia grades was assessed for utility in expression microarray analysis. Cluster analysis and scatter plots of duplicate samples suggest that data of sufficient quality can be obtained from as little as 50 ng of RNA from molecular fixative samples. Taken together, our results show that molecular fixative may be a more versatile substitute for formalin, simultaneously preserving tissue morphology for clinical diagnosis and biomolecules for immunohistochemistry and gene expression analysis.

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“…Formalin fixation offers many advantages: primarily, it is cheap and readily available. Pathologists are routinely trained to examine formalin-fixed tissues (Gugic et al 2007;Srinivasan et al 2002) and…”

Section: Introductionmentioning

confidence: 99%

“…It can cause respiratory irritation and is classified as a carcinogen (Bolt et al 2010;Bosetti et al 2008;Buesa 2008;Gugic et al 2007). Formalin also degrades DNA, RNA, and proteins, which makes formalin-fixed tissues less usable for downstream molecular diagnostics (Buesa 2008;Gugic et al 2007). Formalin is an aldehyde-based fixative that works by cross-linking proteins, which irreversibly damages proteins and nucleic acids (Srinivasan et al 2002).…”

Section: Introductionmentioning

confidence: 99%

“…This is impractical for many private veterinary practices, as these generally lack the facilities to freeze samples at -80˚C and many samples are too small to duplicate samples. Therefore, alternative fixatives have been proposed to allow for both histopathologic examination and molecular diagnostics (Cox et al 2006;Gugic et al 2007;Kap et al 2011;Vincek et al 2003). These have been shown to preserve nucleic acids with results similar to those obtained with fresh or frozen tissues, while still preserving histomorphology.…”

Section: Introductionmentioning

confidence: 99%

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Peer Review #1 of "Comparison of histomorphology and DNA preservation produced by fixatives in the veterinary diagnostic laboratory setting (v0.1)"

2014

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Histopathology is the most useful tool for diagnosis of a number of diseases, especially cancer. To be effective, histopathology requires that tissues be fixed prior to processing. Formalin is currently the most common histologic fixative, offering many advantages: it is cheap, readily available, and pathologists are routinely trained to examine tissues fixed in formalin. However, formalin fixation substantially degrades tissue DNA, hindering subsequent use in diagnostics and research. We therefore evaluated three alternative fixatives, TissueTek® Xpress® Molecular Fixative, modified methacarn, and PAXgene®, all of which have been proposed as formalin alternatives, to determine their suitability for routine use in a veterinary diagnostic laboratory. This was accomplished by examining the histomorphology of sections produced from fixed tissues as well as the ability to amplify fragments from extracted DNA. Tissues were sampled from two dogs and four cats, fixed for 24-48 hours, and processed routinely. While all fixatives produced acceptable histomorphology, formalin had significantly better morphologic characteristics than the other three fixatives. Alternative fixatives generally had better DNA amplification than formalin, although results varied somewhat depending on the tissue examined. While no fixative is yet ready to replace formalin, the alternative fixatives examined may be useful as adjuncts to formalin in diagnostic practices.

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Novel tissue preservative and tissue fixative for comparative pathology and animal research

Cited by 14 publications

References 27 publications

Light microscopic determination of tissue

Light microscopic determination of tissue

Molecular fixative enables expression microarray analysis of microdissected clinical cervical specimens

Peer Review #1 of "Comparison of histomorphology and DNA preservation produced by fixatives in the veterinary diagnostic laboratory setting (v0.1)"

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