Computational protocol: Paper-based electrochemiluminescence sensor for highly sensitive detection of amyloid-β oligomerization: Toward potential diagnosis of Alzheimer's disease

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Protocol publication

[…] Briefly, 5 μL of CSF sample was mixed with SDS sample loading buffer, boiled for 8 min, and separated by 15% SDS-PAGE. Proteins separated in the gels were electrophoretically transferred to PVDF membranes. Membranes were blocked for 1 h using 5% non-fat dry milk in TBS containing 0.5% Tween-20 (TBST) and then incubated with β-amyloid antibody (Santa Cruz, sc-28365) at 4 °C overnight. After three washes with TBST, secondary antibody application was performed at RT for 2 h. The signals were detected with an ODYSSEY Infrared Imaging System (LI-COR). The intensity of the Western blot signals was quantified using ImageJ software (National Institutes of Health (NIH), Bethesda, MD), and the results of densitometric analysis are presented as the ratio of protein/GAPDH protein and are compared with the controls and normalized to 1. [...] The screen-printing fabrication process for the pBPE was performed according to previously reported methods with slight modifications , , . Briefly, the patterning shapes for screen-printing the pBPE chip were designed using Adobe Illustrator CS6 and then delivered to the local screen printing shop to manufacture the screens initially. The fabrication process of the molds used for screen printing and wax-screen printing is shown in Figure . Then, the filter paper was cut into a uniform size. Afterwards, the BPE and driving electrodes were fabricated on the hydrophilic channel by a screen printing technique with carbon ink. Finally, the dried semi-finished product was heated together with the screen at 80 °C for approximately 10 s to melt the wax to produce hydrophobic areas on it with the screen mesh openings, while areas with a cross-linked photosensitive material yielded the hydrophilic channel during the wax-screen printing process. […]

Pipeline specifications

Software tools ImageJ, Adobe Illustrator
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Mus musculus
Diseases Alzheimer Disease