期刊
JOURNAL OF NEUROSCIENCE METHODS
卷 190, 期 1, 页码 49-56出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jneumeth.2010.04.020
关键词
Formalin; Fixation; Immunohistochemistry; Formic acid; Antibodies; Markers; Neuronal; Vascular
资金
- European Commission
- UK MRC
- Department of Health Biomedical Centre scheme
- EURIPIDES
- Medical Research Council [G0600934] Funding Source: researchfish
- MRC [G0600934] Funding Source: UKRI
Human brain tissue is a valuable source of material for research. It is often stored indefinitely in formalin at room temperature which may weaken the immunolabeling with formalin-sensitive antibodies. The present study found that a novel protocol that combined citrate and formic acid pre-treatments with the catalyzed signal amplification (CSA) system was able to recover the lost or weakened immunolabeling with the formalin-sensitive antibodies, anti-CD34, anti-caveolin, anti-P-glycoprotein, anti-neuronal nuclei, anti-parvalbumin, anti-human leukocyte antigen, anti-CD45, anti-CD68 and anti-connexin 43, in post-mortem, human brain tissue that was stored in formalin for up to 10 years at room temperature. Recovered immunolabeling in long-fixed tissue resembled immunolabeling observed in tissue that was fixed for a shorter duration between 6 and 49 days. The findings from this study highlight the importance of testing antibodies for formalin fixation effect prior to studies, especially if long-fixed tissue is used, to enable immunolabeling to be more accurately interpreted. Importantly, this study provides a method of overcoming formalin-masking of antigens in long-fixed human tissue, thus allowing essential immunohistochemical studies to be undertaken using precious human tissue. (C) 2010 Elsevier B.V. All rights reserved.
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