Design and construction of wall-less nano-electrophoretic and nano in micro array high throughput devices for single cell 'omics' single molecule detection analyses
Résumé
Single cell 'omics' requires a technological platform with reliable and high throughput single cell analyses with single molecular detection and quantification. Presently available options are to either to detect many different macromolecules and metabolites extracted from many cells, thus obtaining partial 'omics' of an average cell or to study only few single cells and be limited to semi-quantitative analyses and detection of a few abundant molecules. Here we present a new design and prototype proof of concept construction of high throughput nano-electrophoretic separation (NEA) device and nano in micro array (NiMA) affinity probe device for a complete single cell 'omics' single bio-molecule polymers detection and quantification analyses. Prototype devices were constructed using gallium ion Focus Ion Beam (FIB), Gas Injection System (GIS) and Scanning Electron Microscope (SEM) crossbeam instruments. The NEA device accommodates 100 different cell samplings per 1 cm2 chip with arrays of open nano-electrophoretic guides. The NiMA bio-sensor device on 1 cm2 can accommodate 2500 cells in a micro-well array which consists of 250,000 probe markers in nano-well array located in each micro-well. Using Secondary Ion Mass Spectrometry (SIMS) we have demonstrated the direct detection of a single protein molecule and proved the feasibility of single bio-molecular detection and quantification concept for NEA and NIMA. Our concept validates high throughput and complete and quantitative single cell 'omics' with single molecular detection analyses without labeling. Thus, it is superior to commonly used microfluidics, capillary electrophoresis and micro-arrays using mass spectrometry and fluorescent labeling for molecular detection.
Mots clés
Biomolecules
Capillary electrophoresis
Ion beams
Ions
Mass spectrometry
Molecules
Nanosensors
Probes
Scanning electron microscopy
Secondary ion mass spectrometry
Spectrometry
Detection and quantifications
Electrophoretic separations
FIB
High-throughput analysis
Quantification
Semi-quantitative analysis
Single cells
Single molecular detections
Throughput