Biofunctionalization as Key Step in Microfluidic Based Applications

Transcription

1 MicroNano Conference, December , Amsterdam Biofunctionalization as Key Step in Microfluidic Based Applications Dr. Guido Bared Dr. Wilfried Weigel Scienion AG, Berlin

2 Scienion - Company Background Spin off from Max-Planck Institute of Molecular Genetics in Employees Core competence Arrays/Spot-Based Analytics Biofunctionalization of microfluidic chips, biosensor surfaces, MTP Key technology Non contact printing of picoliter volumes Portfolio sciflexarrayer, scanner, consumables and services

3 Spots on Microarrays and Biosensors Microarrays arrays of spots in a pre-defined area Biosensors arrays of spots exactly at a predefined position Fluorescence scanner image of an array on a glass slide Colorimetric scanner image of an array in a well of a microplate CCD Images of the loaded electrodes of a CMOS sensor

4 Spot-Based Analytics in µm Range ( µm) Spots µm Target Capture Probe immobilized capture molecules interact with labeled target/analyte from sample solution Capture Probes oligonucleotides, antibodies, antigens, carbohydrates, peptides, Targets/Analytes RNA, DNA, antibodies, proteins

5 Detection Methods Fluorescence scanner image of an array on a glass slide Colorimetric scanner image of an array in a well of a microplate Chemiluminescent image of an array in a well of a microplate Labelfree detection - Electrochemical readout - Surface plasmon resonance SPR - Mass spectrometry

6 PDC-Piezo activation pl µm spots

7 How to create Spots?

8 Tuning of Droplets Speed and volume of ejected droplets can be modified by variation of pulse parameters pulse form, voltage, pulse width and frequency Volt Speed Duration [µs] Volume pl

9 Flexibility in Ultra Low Volume Dispensing Different Printing Patterns Spots Lines 120 micron Bars/ Fields/ Coatings 720 x 4,100 micron Variety of Substrates Microplates Microwells Electronic Chips Biosensors

10 How to get the Spots on the Substrates?

11 Scalable Technology From R&D to High-Throughput Production RESEARCH & DEVELOPMENT SMALL MEDIUM SCALE LARGE SCALE ADDRESSING DIVERSE NEEDS WITH ONE TECHNOLOGY Capacity Precision Speed Accuracy Upgradeability Reliability Space Integration Increasing Throughput

12 12 High Quality Arrays by Application of Coated Nozzles Protein Array PDC 70 Standard Protein Array PDC 70 Coated

13 Surface Design to Immobilize a Capture Probe

14 Large Variety of Combinations of Support and Content Supports 1 x 3 slides 3 x 5 plates Microplates Membranes CDs Biosensors Cassettes Custom Glass, Silicon oxide, Silicon nitride Polymers Gold Content Nucleic Acids Proteins (Including Antibodies) Peptides Intact Cells Tumor/ Tissue Lysates Aqueous/ Organic Solvent based Nanoparticles Monomers

15 From 2D to 3D Functional Surfaces 2D Functional Layers 3D Polymer Layers 3D Hydrogels functional group hydrophilic group Spacer-Function dry swollen Copolymers of functional and hydrophilic monomers Hydrophilic polymer networks, e.g. tween derivative crosslinked with polyethylene glycole Polysiloxane layers on glass The Standard microarray surface Tailor made functional density, hydrophilicity and introduction of PEG spacer Provides solution-like environment to preserve protein biofunctionality

16 Individual Materials Properties Determine Method of Surface Functionalization Supports of different materials and geometries glass/silicon polymers gold microplates, membranes, polymers, biosensors, microfluidic devices 2D functional layers 3D layers from plasma Self Assembling from silanization treatment, photo- Monolayers (SAMs) polymerization from thiols O Si O Si O Si O O O O

17 Immobilization on Non Activated Surfaces

18 Immobilization Using a Photoreactive Polymer Printing of drops ( pl) containing capture probe and soluble polymer 100 µm DMAA MABP SSNa DNA-Sonde Capture Probe (DNA) Copolymer Polymer Verknüpfungspunkt Crosslink point

19 Fast Way of Capture Probe Immobilization Using Polymer Matrices Workflow Preparation of Samples (polymer + capture probe) Non Contact Printing UV crosslinking (2 min) Immobilization on non activated surfaces Immobilization on substrates with protein resistant surfaces Immobilization of capture probes to polymer network and attachment of probe/polymer network to substrate scipoly 3D Immobilization Technology

20 scipoly3d Scienion Application Examples Oligonucleotide Arrays scichip Epoxy scipoly3d Immobilizaton Control Hybridization Control

21 Quick Methods for Biofunctionalization on Polymer Surfaces Classical approach for surface activation, printing immobilization Surface Functionalization Printing Probes 24 hours < 1 hour scipoly3d Technology Printing scipoly3d/probes UV- Immobilization < 1 hour 2 min Immobilization on non activated polymer surfaces as microfluidic chips

22 Highthroughput Inline Production Systems for Surface Modification, Printing and Immobilization Feature Specification Nozzles Up to mm spacing Independent Z Yes Slides 4* Micro Plates 1* Source-plate Holder 2 (96, cool/heat) Humidity Control Up to 75% Power 110V/230V 60Hz/50Hz Weight/Portal 700 Lbs Dimensions (LxWxH) 4.3 x 2.6 W x 3.9 H Resolution 1µm Accuracy (< ±20µm) Precision (< ±5µm) Max Speed 3m/s

23 Drops Under Oil

24 Thank you very much for your attention Dr. Guido Bared Scienion AG, Volmerstr. 7b, Berlin Tel. +49 (0) Fax bared@scienion.de

25 DNA Microarrays for Detection of Microbiological Freshwater Quality Detection of pathogenic and toxic organisms RNA isolation from field samples and hybridization on oligonucleotide microarrays of 250 probes EU FP7 microaqua project,

26 Singleplex Multiplex Analysis 1x1 2x2 3x3 4x4 5x5 6x6 7x7 8x8 9x9 10x10 11x11 12x12 Traditional ELISA one analyte per well Multiplex ELISA multiple analytes per well

27 Application of scimultiplex Technology Diagnostic HPV Typing Array in sciplexplate 96

28 Microarrays for Material Research

29 Nanomaterials material research Characterization of by TEM TEM (Transmission Electron Microscopy) Grid Holder TEM not just for NP characterization! Also for material research, biology, etc..

30 Nanomaterials Loading of Substrates for Transmission Electron Microscopy (TEM) scitem Application of picoliter printing allows precise of samples on the highly sensitive support films Technology allows loading of one TEM windows with multiple samples 1x1 mm silicon nitride film (50 nm thick) after printing of 50 pl drops in a 10x10 pattern TEM images of 5-7 nm Ni Nanoparticles

31 Tools and Service for Microarray Based Application Products sciflexarrayer sciplexplate scichip scibuffer Key Technologies Contact Free Spotting Surface Functionalization Microarray Applications Services Product/ Process Development & Manufacturing

32 Microarray Platforms Different Levels of System Integration Modular systems Slides scichip Microplates Integrated systems Chips carrying the arrays incorporated in Microfluidic Cartridges sciplexplate Workstations load & go Automation Genomica, SQI,