Fixation of Tissues

Grizzle, William E.; Fredenburgh, Jerry; Myers, Richard S.

doi:10.1016/b978-0-443-10279-0.50011-7

Cited by 44 publications

(38 citation statements)

References 39 publications

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“…29 Additionally, interlaboratory variation in tissue processing may affect final patterns of tissue shrinkage. 35 The study reported here was performed with samples obtained from recently euthanized cats. All skin samples were collected within 30 minutes after cats were euthanized.…”

Section: Discussionmentioning

confidence: 99%

Effect of histologic processing on dimensions of skin samples obtained from cat cadavers

Jeyakumar¹,

Smith²,

Schleis³

et al. 2015

ajvr

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OBJECTIVE To determine changes in dimensions of feline skin samples as a result of histologic processing and to identify factors that contributed to changes in dimensions of skin samples after sample collection. SAMPLE Cadavers of 12 clinically normal cats. PROCEDURES Skin samples were obtained bilaterally from 3 locations (neck, thorax, and tibia) of each cadaver; half of the thoracic samples included underlying muscle. Length, width, and depth were measured at 5 time points (before excision, after excision, after application of ink to mark tissue margins, after fixation in neutral-buffered 10% formalin for 36 hours, and after completion of histologic processing and staining with H&E stain). Measurements obtained after sample collection were compared with measurements obtained before excision. RESULTS At the final time point, tissue samples had decreased in length (mean decrease, 32.40%) and width (mean decrease, 34.21%) and increased in depth (mean increase, 54.95%). Tissue from the tibia had the most shrinkage in length and width and that from the neck had the least shrinkage. Inclusion of underlying muscle on thoracic skin samples did not affect the degree of change in dimensions. CONCLUSIONS AND CLINICAL RELEVANCE In this study, each step during processing from excision to formalin fixation and histologic processing induced changes in tissue dimensions, which were manifested principally as shrinkage in length and width and increase in depth. Most of the changes occured during histologic processing. Inclusion of muscle did not affect thoracic skin shrinkage. Shrinkage should be a consideration when interpreting surgical margins in clinical cases. 945).

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Section: Discussionmentioning

confidence: 99%

Effect of histologic processing on dimensions of skin samples obtained from cat cadavers

Jeyakumar¹,

Smith²,

Schleis³

et al. 2015

ajvr

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“…Second, to fix and preserve the biological composition of sampled Symbiodinium and prevent the formation of artificial chemical bonds during the sampling process, a coagulant (precipitating) fixative, TCA, was used. Coagulant fixatives reduce the solubility of protein molecules by disrupting hydrophobic interactions yet do not covalently bind to proteins as do the cross-linking, aldehyde-based fixatives [11]. Third, for spectral analyses, the amide II band (A 1510 ) of each spectrum was used as the internal control for qualitative and quantitative analysis (electronic supplementary material, figure S3).…”

Section: Resultsmentioning

confidence: 99%

Assessment of metabolic modulation in free-living versus endosymbiotic Symbiodinium using synchrotron radiation-based infrared microspectroscopy

et al. 2011

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The endosymbiotic relationship between coral hosts and dinoflagellates of the genus Symbiodinium is critical for the growth and productivity of coral reef ecosystems. Here, synchrotron radiationbased infrared microspectroscopy was applied to examine metabolite concentration differences between endosymbiotic (within the anemone Aiptasia pulchella) and free-living Symbiodinium over the light-dark cycle. Significant differences in levels of lipids, nitrogenous compounds, polysaccharides and putative cell wall components were documented. Compared with free-living Symbiodinium, total lipids, unsaturated lipids and polysaccharides were relatively enriched in endosymbiotic Symbiodinium during both light and dark photoperiods. Concentrations of cell wallrelated metabolites did not vary temporally in endosymbiotic samples; in contrast, the concentrations of these metabolites increased dramatically during the dark photoperiod in free-living samples, possibly reflecting rhythmic cell-wall synthesis related to light-driven cell proliferation. The level of nitrogenous compounds in endosymbiotic cells did not vary greatly across the light-dark cycle and in general was significantly lower than that observed in free-living samples collected during the light. Collectively, these data suggest that nitrogen limitation is a factor that the host cell exploits to induce the biosynthesis of lipids and polysaccharides in endosymbiotic Symbiodinium.

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“…Formalin fixation results in the formation of protein-nucleic acid and protein-protein crosslinks within the cell, limiting the movement of biomolecules and organelles, and preserving the tissue from decay (Grizzle et al, 2008). The most widely used fixation protocol includes immersion of the sample in 10% buffered formalin (final concentration of formaldehyde 3.7% in phosphate buffered saline) for 24 hours.…”

Section: Protein Modifications Induced By For-maldehyde Fixationmentioning

confidence: 99%

Proteomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Samples: Pitfalls and Potentials

Valera¹,

Walter²,

Linehan³

et al. 2009

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Proteomic analysis of clinical samples has been traditionally performed using fresh or frozen tissues. However, millions of embedded samples are stored worldwide in pathology repositories with linked long-term outcome data. Such samples are believed to be unsuitable for proteome analysis because of the extensive protein modification attained during tissue processing. Formaldehyde, in the form of buffered formalin, is the most commonly used fixative during processing before embedding in paraffin. These formalin-fixed paraffin embedded (FFPE) tissue samples are widely used in pathology labs for qualitative protein analysis using immunohistochemistry (IHC). For this purpose, FFPE tissues are treated in different ways in order to reverse chemical modifications by formaldehyde that limit protein antigen detection by specific antibodies. Accordingly, intensive research efforts are now attempting to adapt the sample pretreatments used in immunohistochemistry to facilitate high throughput proteome analysis of FFPE tissues. Two such approaches have been successful: a liquid-based proteomic approach and an in situ approach, mainly based on separation and identification technologies such as liquid chromatography and mass spectrometry. These efforts have proven that similar protein profiles and numbers of polypeptides can be attained as using frozen sections. In this article, a review is provided of methods and current trends for the extraction of peptides and proteins from FFPE tissues for use proteomic analysis. Limitations in these methods and future directions to overcome these are also outlined. Ongoing improvements in protein extraction technology and analysis techniques will enable identification of novel disease biomarkers from FFPE tissues.

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Fixation of Tissues

Cited by 44 publications

References 39 publications

Effect of histologic processing on dimensions of skin samples obtained from cat cadavers

Effect of histologic processing on dimensions of skin samples obtained from cat cadavers

Assessment of metabolic modulation in free-living versus endosymbiotic Symbiodinium using synchrotron radiation-based infrared microspectroscopy

Proteomic Analysis of Formalin-Fixed Paraffin Embedded (FFPE) Samples: Pitfalls and Potentials

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