Effects of Long-Term Storage on the Detection of Proteins, DNA, and mRNA in Tissue Microarray Slides

Karlsson, Christina; Karlsson, Mats G.

doi:10.1369/0022155411423779

Cited by 53 publications

(43 citation statements)

References 25 publications

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“…59 Published methods to reduce antigen decay in stored tissue sections include storage in a vacuum, covering slides with a layer of paraffin, addition of wax-soluble antioxidants to paraffin, storage of sections in a nitrogen chamber, immersion of deparaffinized slides in stabilizing solutions at 4 C, and optimizing fixation and antigen retrieval methods. 5,6,10,21,22,50,51 In our laboratory, inconsistent immunoreactivity of certain biomarkers in stored paraffin sections prompted an examination of the antigenic stability of 49 biomarkers (infectious agents and cell antigens) used in diagnostic pathology in FFPE tissue sections stored at room temperature (RT) or 4 C in the dark, or at RT with exposure to sunlight or fluorescent light.…”

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confidence: 99%

Immunohistochemical Evaluation of the Effects of Paraffin Section Storage on Biomarker Stability

et al. 2013

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Environmental stresses can alter immunoreactivity of biomarkers in stored tissue sections. The effect of temperature and lighting on 49 cellular or microbial antigens was evaluated in 4 serial paraffin sections, cut 12 months, 10 months, 8 months, 5 months, 3 months, 1 month, 3 days, and 1 day before immunohistochemistry. Slides were stored at room temperature (RT) in the dark, at 4°C in the dark, at RT under fluorescent light, or at RT with windowpane exposure to sunlight. Immunohistochemistry was performed simultaneously in an automated immunostainer. Immunoreactivity was compared with that in the corresponding 1-day-old section and scored as 4 (<10% reduction), 3 (10%-25% reduction), 2 (26%-60% reduction), 1(>60% reduction), or 0 (no reactivity). Any loss of immunoreactivity was proportional to the tissue section age and was least in sections stored in the dark. Immunoreactivity was only completely lost in light-exposed sections and as early as 1 month for CD45. Other markers with complete loss of immunoreactivity were bovine viral diarrhea virus, CD18 (only with fluorescent light), CD31, CD68, canine parvovirus, chromogranins, and thyroid transcription factor-1. Markers with complete loss after light exposure also had reduced immunoreactivity when stored in the dark, as early as day 3. Eight markers (Bartonella spp, CD11d, high molecular weight cytokeratins, feline coronavirus, GATA-4, insulin, p63, progesterone receptor) had minimal decrease in immunoreactivity, regardless of treatment. In conclusion, light-induced antigen decay (tissue section aging) is antigen dependent and could explain unexpectedly weak or negative immunohistochemical reactions in stored paraffin sections.

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Immunohistochemical Evaluation of the Effects of Paraffin Section Storage on Biomarker Stability

et al. 2013

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“…It has previously been reported that choice of Ab and use of methodology are of significant importance. The community should be aware of the multitude of issues that influence the staining outcome (fixative, fixation time, storage time of sections) and that this influence varies from one epitope to another (McNicol and Richmond 1998;Shi et al 2001;Gelpi et al 2007;D'Amico et al 2009;Pikkarainen et al 2010a;Karlsson and Karlsson 2011;Kovacs et al 2012). Another issue that the BNE consortium has become aware of is the need for precise detailed instructions.…”

Section: Discussionmentioning

confidence: 99%

“…IHC methodology is prone to considerable variability that might cause differing results, as was elegantly reported by Mackenzie et al (2006b). Today, it is acknowledged that use of the IHC methods requires knowledge and competence both regarding methodology and interpretation of the result obtained (McNicol and Richmond 1998;Shi et al 2001;Gelpi et al 2007;D'Amico et al 2009;Pikkarainen et al 2010a; Karlsson and Karlsson 2011;Kovacs et al 2012). In addition, the agreement rate for the designation of a specific type of a disease by numerous investigators might be less optimal, as previously reported by BrainNet Europe (BNE) (Alafuzoff et al 2008b(Alafuzoff et al , 2009a.…”

Section: Introductionmentioning

confidence: 99%

Neuropathological assessments of the pathology in frontotemporal lobar degeneration with TDP43-positive inclusions: an inter-laboratory study by the BrainNet Europe consortium

et al. 2014

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The BrainNet Europe consortium assessed the reproducibility in the assignment of the type of frontotemporal lobar degeneration (FTLD) with TAR DNA-binding protein (TDP) 43 following current recommendations. The agreement rates were influenced by the immunohistochemical (IHC) method and by the classification strategy followed. p62-IHC staining yielded good uniform quality of stains, but the most reliable results were obtained implementing specific Abs directed against the hallmark protein TDP43. Both assessment of the type and the extent of lesions were influenced by the Abs and by the quality of stain. Assessment of the extent of the lesions yielded poor results repeatedly; thus, the extent of pathology should not be used in diagnostic consensus criteria. Whilst 31 neuropathologists typed 30 FTLD-TDP cases, inter-rater agreement ranged from 19 to 100 per cent, being highest when applying phosphorylated TDP43/IHC. The agreement was highest when designating Type C or Type A/B. In contrast, there was a poor agreement when attempting to separate Type A or Type B FTLD-TDP. In conclusion, we can expect that neuropathologist, independent of his/her familiarity with FTLD-TDP pathology, can identify a TDP43-positive FTLD case. The goal should be to state a Type (A, B, C, D) or a mixture of Types (A/B, A/C or B/C). Neuropathologists, other clinicians and researchers should be aware of the pitfalls whilst doing so. Agreement can be reached in an inter-laboratory setting regarding Type C cases with thick and long neurites, whereas the differentiation between Types A and B may be more troublesome.

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“…Factors such as cold ischemia time in surgical setting, post-mortem delay (PMD), fixation time (FT), paraffin, storage time in paraffin, storage temperature, age of the cut sections, antigen retrieval (AR) technique and DS have reported to influence the outcome of IHC (Pikkarainen et al 2010; Skaland et al 2010; Karlsson and Karlsson 2011; Engel and Moore 2011; Ramos-Vara and Miller 2014; Grillo et al 2015, 2017; Lundström et al 2018). Furthermore, in the setting of post-mortem (PM) tissue, not only PMD but also agonal state influences the IHC (Monoranu et al 2009; Lundström et al 2018).…”

Section: Introductionmentioning

confidence: 99%

Characteristics of the tissue section that influence the staining outcome in immunohistochemistry

Libard

Cerjan

Alafuzoff

2018

Histochem Cell Biol

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Immunohistochemistry (IHC) is influenced by several factors such as cold ischemia time, fixative, fixation time, paraffin, storage time, antibody, antigen retrieval technique and detection systems. In the setting of post-mortem tissue, not only post-mortem delay, but also agonal state is of interest. Here, we assessed an additional variable, i.e., the thickness of the section, and noted that this variable also influenced the IHC outcome. This is of significance when the extent of labelling is a parameter to be assessed, for example when assigning a stage or grade of a disease. Furthermore, when assessing brain tissue with neurons, soma measuring from 4 to 100 µm, various cellular compartments composed of different proteins are localised in sections measuring 4 or 7 µm. Thus, what is seen in a 7-µm-thick section might be lacking in a 4-µm-thick section. Lack of information regarding the molecular size of commercial antibodies is also disturbing as this parameter might influence the distribution of the molecule in the three-dimensional section. The choice of antibody to be used and the staining methodology have been acknowledged being of significance for IHC outcome; however, neither sections thickness or the molecular weight has been discussed sufficiently. IHC has been shown to be an unpredictable technique used for assessment of tissue. This emphasises the need for detailed methodological descriptions in publications, the need to acknowledge and to harmonize all eventual pitfalls related to this methodology.

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Effects of Long-Term Storage on the Detection of Proteins, DNA, and mRNA in Tissue Microarray Slides

Cited by 53 publications

References 25 publications

Immunohistochemical Evaluation of the Effects of Paraffin Section Storage on Biomarker Stability

Immunohistochemical Evaluation of the Effects of Paraffin Section Storage on Biomarker Stability

Neuropathological assessments of the pathology in frontotemporal lobar degeneration with TDP43-positive inclusions: an inter-laboratory study by the BrainNet Europe consortium

Characteristics of the tissue section that influence the staining outcome in immunohistochemistry

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