Jump to main content
Jump to site search

Issue 5, 2017
Previous Article Next Article

High-selectivity cytology via lab-on-a-disc western blotting of individual cells

Author affiliations

Abstract

Cytology of sparingly available cell samples from both clinical and experimental settings would benefit from high-selectivity protein tools. To minimize cell handling losses in sparse samples, we design a multi-stage assay using a lab-on-a-disc that integrates cell handling and subsequent single-cell western blotting (scWestern). As the two-layer microfluidic device rotates, the induced centrifugal force directs dissociated cells to dams, which in turn localize the cells over microwells. Cells then sediment into the microwells, where the cells are lysed and subjected to scWestern. Taking into account cell losses from loading, centrifugation, and lysis-buffer exchange, our lab-on-a-disc device handles cell samples with as few as 200 cells with 75% cell settling efficiencies. Over 70% of microwells contain single cells after the centrifugation. In addition to cell settling efficiency, cell-size filtration from a mixed population of two cell lines is also realized by tuning the cell time-of-flight during centrifugation (58.4% settling efficiency with 6.4% impurity). Following the upstream cell handling, scWestern analysis detects four proteins (GFP, β-TUB, GAPDH, and STAT3) in a glioblastoma cell line. By integrating the lab-on-a-disc cell preparation and scWestern analysis, our platform measures proteins from sparse cell samples at single-cell resolution.

Graphical abstract: High-selectivity cytology via lab-on-a-disc western blotting of individual cells

Back to tab navigation

Supplementary files

Publication details

The article was received on 27 Oct 2016, accepted on 05 Jan 2017 and first published on 19 Jan 2017


Article type: Paper
DOI: 10.1039/C6LC01333C
Citation: Lab Chip, 2017,17, 855-863
  •   Request permissions

    High-selectivity cytology via lab-on-a-disc western blotting of individual cells

    J. J. Kim, E. Sinkala and A. E. Herr, Lab Chip, 2017, 17, 855
    DOI: 10.1039/C6LC01333C

Search articles by author

Spotlight

Advertisements