A novel method for reliable nuclear antibody detection in tissue with high levels of pathology-induced autofluorescence

作者：Spanswick Simon C^*; Bray Doug; Zelinski Erin L; Sutherland Robert J

来源：Journal of Neuroscience Methods, 2009, 185(1): 45-49.

DOI：10.1016/j.jneumeth.2009.09.007

• Immunohistochemistry
• Adrenalectomy
• Stroke
• Neurogenesis

Immunofluorescence is the basis for many techniques used to quantify phenomena in neuroscience research, in both normal and pathological tissue. Autofluorescence (non-specific, broad spectrum background fluorescence) is an unfortunate consequence of damage to brain tissue. Damage-induced autofluorescence potentially confounds analyses of tissue labeled with fluorescent markers in many experiments. This is especially problematic in protocols that utilize co-localization methods such as BrdU/NeuN in which autofluorescence might lead to overestimates of the number of double-labeled cells. Techniques to reduce autofluorescence are variable and relatively ineffective in damaged brain tissue. Here we show using confocal microscopy that damage-induced autofluorescence does not colocalize with the nuclear specific markers DAPI or Hoechst. Further co-localization of nuclear markers such as Ki67 or BrdU/NeuN with DAPI or Hoechst should serve to help discriminate between intended and spurious fluorescent signal.

• 出版日期2009-12-15