development of an electron microbeam for cell culture studies
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Development of an Electron Microbeam for Cell Culture Studies. T. W. Botting, L. A. Braby, and J. R. Ford Texas A&M University. Overview. Background Construction Operation Current Experiments Future. Objective. - PowerPoint PPT PresentationTRANSCRIPT
Development of an Electron Development of an Electron Microbeam for Cell Culture StudiesMicrobeam for Cell Culture Studies
T. W. Botting, L. A. Braby, and J. R. Ford
Texas A&M University
OverviewOverview
BackgroundConstructionOperationCurrent ExperimentsFuture
ObjectiveObjective
Our main objective is to achieve a better understanding of the risk to human health due to everyday exposure to low doses of ionizing radiation.
Most occupational and public radiation exposures are due to x and rays
so
concern is about the effects of small numbers of moderate energy electrons
(10 to 1000 keV)
How do we study this directly?How do we study this directly?
– Need source for low-to-moderate energy electrons
– Need method to deliver them exactly where desired
We have used an electron microbeam to try to quantify bystander effects produced by moderate energy electrons
beam delivery of electron dosebeam delivery of electron dose
Targeting• irradiation paths• discrete locations
Dose• duration• intensity
Energy
Electron Beam ProductionElectron Beam Production
Electron source• low-power tungsten filament• low voltage power supply• isolation transformer
Accelerator Tube• custom-made 3-section ceramic• equipotential rings• high voltage power supply
Beam DeliveryBeam Delivery
Collimator Assembly• capillary tube• swivel mounts for alignment
Cell dish stage• x-y motion control
Microscope and camera• targeting
Electron Microbeam ApparatusElectron Microbeam Apparatus
Less than 4 feet high
Capillary-style collimator
Accelerator tube up to 100,000 Volts to produce up to 100keV electrons
Source and AcceleratorSource and Accelerator
- Source- Source
- Accelerator tube- Accelerator tube
Voltage dividers -Voltage dividers -
\\Faraday Cup controlFaraday Cup control
Turbo pump -Turbo pump - Equipotential ringsEquipotential rings//
3D Schematic3D Schematic
Collimator Stand and MicroscopeCollimator Stand and Microscope
X-Y motion controlX-Y motion control||
CCD camera -CCD camera -
- Stage- Stage
\\ Capillary CollimatorCapillary Collimator
Light SourceLight Source//
Cell culture dishesCell culture dishes
Final Construction DetailsFinal Construction Details
Voltage dividers• 30 M per tube section for smooth gradient
Exit collimation• 5m and 300m exit aperatures
Exit window• 2m thick mylar (same as cell dishes)
OperationOperation
Electron source• provides electron beam up to 1 nanoamp on
the Faraday cup
Stable at up to 85 kV so far• beams at up to 90kV
Software control of targeting• line traces• discrete spots
Desired ImprovementsDesired Improvements
Beam stabilityBeam currentBeam transmission
Bystander Effect ExperimentsBystander Effect Experiments
Irradiate nearly confluent cellsCDKN1A and PCNA versus distance
• AG 1522 human fibroblasts• Clone 9 rat liver line• RIE mouse intestine line• HBEC human primary bronchial cells
Micronuclei assay• AG 1522 human fibroblasts
Some Future Directions…Some Future Directions…
Further micronuclei assays• Clone 9 rat liver line• RIE mouse intestine line• HBEC human primary bronchial cells
NTEC Rat primary tracheal cells• All three methods (CDKN1A, PCNA, micronuclei)
Complete comparison matrix with our positive ion beam results as a control