Our Process

The Optofluidic System

The Beacon is a platform for single cell culture that uses light to move cells via Opto-Electric Positioning (OEP).

White LED light generates a small electric field across the photo transitor grid of the chip to move nano objects
Using cell type specific cell detection algorithms, OEP can be used to manipulate hundreds of cells within a field of view (FOV) and sort individual cells into isolated nanopen compartments

Recovery of Cells using OEP Export

OEP can be used to move cells out of nanopen compartments during export. Recovered cells are exported in 5-10 uL fluidic packages to onboard well plate incubators or tubes.

Cells can be exported either:

live and incubated for expansion in culture media
lysed and chilled to support nucleic acid recovery

Culture and Manipulation of Cells on the Beacon Chips

There are varied sizes of chips that have varied sizes of nanopens that correspond to the intended cell type. We can work with dividing and/or adherent cells, T-Cells, and B-cells.

The sample types that we have worked with so far include:

mouse plasma B cells prepared from spleens of immunized mice
rabbit memory B cells prepared from PBMCs of immunized rabbits
sorted human T-cells
adherent cells (CHO)
llama memory B cells prepared from PBMCs of llamas

Microfluidic Chip Assays of Single Cells

Multiplexed specificity antibody secretion assays

use up to five soluble antigens to determine the specificity of secreted antibodies for antigens of interest
each assay takes ~1 hour
can be automated

Targeted Pen Selection (TPS) allows enrichment for cell types of interest based on physical properties or fluorescent markers. Distinct populations of cells can be identified during import while cells are in the main channel. TPS can be used seed pens in either GFP+ or RFP+ states, allowing distinct populations to be identified, captured, isolated, and tracked.

bifhr

Optics and Imaging Capabilities

The Beacon houses a single laser source (Class 4, 808 nm wavelength, max 4W optical power, continuous wave) contains five different filter cubes, and can capture fluorescence and bright-field time-lapse images giving us the capability of designing multiple types of assays.

Assay Scoring

Onboard software allows us to analyze images with nanopen resolution. Automated scoring detects and labels positive pens by identifying the fluorescent signal ('bloom') in the main channel immediately above the selected pen.

All automated pen selections undergo manual verification to ensure the sensitivity and accuracy of assay scoring. Supporting analysis software allows all data to be aggregated and pens with desired specificity between multiple assays can be tabulated by sample type, location, cell count, etc. These visualizations inform which cells will be exported off of the Beacon and into a 96-well plate, which then can be cloned for sequence recovery.

Our Process: From Beacon to Bench