Quantitative quadruple-label immunofluorescence of mitochondrial and cytoplasmic proteins in single neurons from human midbrain tissue
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- 作者：Anne Grü ; newalda ; b ; anne.gruenewald@newcastle.ac.uk" class="auth_mail ; Nichola Z. Laxa ; nichola.lax@newcastle.ac.uk" class="auth_mail ; Mariana C. Rochaa ; m.rocha@newcastle.ac.uk" class="auth_mail ; Amy K. Reevea ; amy.reeve@newcastle.ac.uk" class="auth_mail ; Philippa D. Hepplewhitea ; philippa.hepplewhite@newcastle.ac.uk" class="auth_mail ; Karolina A. Rygiela ; karolina.rygiel@newcastle.ac.uk" class="auth_mail ; Robert W. Taylora ; Robert.taylor@newcastle.ac.uk" class="auth_mail ; Doug M. Turnbulla ; doug.turnbull@newcastle.ac.uk" class="auth_mail
- 关键词：Mitochondria ; Complex I ; Complex IV ; Single cell analysis ; Midbrain neurons ; Immunofluorescent labelling
- 刊名：Journal of Neuroscience Methods
- 出版年：30 July 2014
- 年：2014
- 卷：232
- 期：Complete
- 页码：143-149
- 全文大小：2007 K
文摘
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We developed an assay to quantify respiratory chain deficiencies in single neurons.
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Quadruple-label immunofluorescence was combined with quantitative image analysis.
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The single-cell assay was applied to tyrosine hydroxylase-positive midbrain neurons.
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The expression of complexes I and IV was determined relative to mitochondrial mass.
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The assay proved specific in patients with known respiratory chain deficiencies.