1 / 30

State-of-the-art probes

State-of-the-art probes. Alan Bigelow. Alternative sensing methods Real-time, single-cell analysis techniques. Outline. Miniature ion-selective single-cell probes Collaboration with the Biocurrents Research Lab at Woods Hole Probe positioner and manipulator

mave
Download Presentation

State-of-the-art probes

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. State-of-the-art probes Alan Bigelow • Alternative sensing methods • Real-time, single-cell analysis techniques

  2. Outline • Miniature ion-selective single-cell probes Collaboration with the Biocurrents Research Lab at Woods Hole • Probe positioner and manipulator • Laser excited single-cell optical nanosensors Collaboration with Tuan Vo-Dihn • Kambiz Pourrezaei collaboration • A Surface-Enhanced Raman Scattering Nano-Needle for Cellular Measurements • Carbon Nanotube Cellular Endoscopes • Automated Microscope Observation Environment for Biological Analyses (AMOEBA)

  3. Miniature Ion-Selective Single-Cell Probes These probes are used to study changes of inflows or outflows of small molecules from individual living cells, in response to spatially-defined damage 1 mm 1 mm 1mm

  4. Making Probes

  5. Laser-Based Micropipet Pulling Device (Model P-2000; Sutter Industries)

  6. Glass Microelectrode Copper Wire Graphite Epoxy Paste Epoxy Carbon Fiber O-Phenylenediamine Nafion The Woods-Hole team have developed sensors for a variety of molecules, such as nitric oxide:

  7. Getting these single-cell probes into position, efficiently and reproducibly....A non-trivial task!

  8. Offset Hinge: probe positioning system

  9. Other manipulations using the offset hinge mount • Cell micro-injection • Single cell harvesting • Optical fiber based Raman spectroscopy • Orientation of medaka embryos

  10. Nanobiosensors Collaboration with Tuan Vo-Dinh Advanced Biomedical Science and Technology Group Life Science Division Oak Ridge National Laboratory

  11. Nano-biosensor tip • Pulled nano-sensors have tip diameters of approximately 40-50 nm • Final coated fibers are approximately 200 nm diameter • Antibody coated tips for specificity in binding • Nanometer diameter tip provides near-field excitation Sensor inside cell

  12. Metalic coating of probe end to prevent leakage of the excitation light Gold, Aluminum, or Silver

  13. Scanning Electron Microscope Images of a Nanofiber Before Metal Coating (tip diameter ~50nm) After Metal Coating (tip diameter 250-300nm)

  14. Nano-probe attachment

  15. Automated Microscope Observation Environment for Biological Analyses (AMOEBA)

  16. Environment Control User Requests: Physiological conditions Control temperature (e.g. 37 ± 0.5 ºC) Control medium concentrations (CO2, pH, oxygen, etc.) Initial Solutions: • Air-CO2 mixture: allows accurate particle count; limited time • Heater ring: Maintains temperature; cell medium evaporates

  17. AMOEBA Flow system for temperature-controlled medium exchange Flexible, user-friendly, modular design offers: • Medium aspiration, replacement, and collection • Multiple dispensers to change medium type during experiment • Additive introduction, such as trypsin to remove cells • Sensor insertion to monitor absorbed gas • Microfluidics compatibility: Lab-on-a-chip for in-line analysis

  18. “Flow” Diagram Example Reservoir I Additive Inlet Reservoir II Pump Heater / Cooler Reservoir III Hinge mount Microbeam Dish Lab-on-a-chip Dispenser

  19. Proof of Principle Cells were observed for 2 hours with circulating medium at 37 ± 0.5 ºC.

  20. Proof of Principle System included heated-window cap, to assist heating control.

  21. Lab-in-a-Box Sensor • Assemble your own system from modules. • Automation is computer controlled. • AMOEBA is flexible and has potential use in labs across the country and the world.

More Related