A Variable-Energy Soft X-Ray Microprobe to Investigate Mechanisms of the Radiation-Induced Bystander Effect.
The Gray Cancer Institute has pioneered the use of X ray focussing techniques to develop systems for micro irradiating individual cells and sub cellular targets in vitro. Cellular micro irradiation is now recognised as a highly versatile technique for understanding how ionising radiation interacts with living cells and tissues. The strength of the technique lies in its ability to deliver precise doses of radiation to selected individual cells (or sub cellular targets). The application of this technique in the field of radiation biology continues to be of great interest for investigating a number of phenomena currently of concern to the radiobiological community. One important phenomenon is the so called ‘bystander effect’ where it is observed that unirradiated cells can also respond to signals transmitted by irradiated neighbours. Clearly, the ability of a microbeam to irradiate just a single cell or selected cells within a population is well suited to studying this effect. Our prototype ‘tabletop’ X-ray microprobe was optimised for focusing 278 eV C-K X rays and has been used successfully for a number of years. However, we have sought to develop a new variable energy soft X-ray microprobe capable of delivering focused CK (0.28 keV), Al-K (1.48 keV) and notably, Ti-K (4.5 keV) X rays. Ti-K X rays are capable of penetrating several cell layers and are therefore much better suited to studies involving tissues and multi cellular layers. In our new design, X-rays are generated by the focussed electron bombardment of a material whose characteristic-K radiation is required. The source is mounted on a 1.5 x 1.0 metre optical table. Electrons are generated by a custom built gun, designed to operate up to 15 kV. The electrons are focused using a permanent neodymium iron boron magnet assembly. Focusing is achieved by adjusting the accelerating voltage and by fine tuning the target position via a vacuum position feedthrough. To analyze the electron beam properties, a custom built microscope is used to image the focussed beam on the target, through a vacuum window. The X-rays are focussed by a zone plate optical assembly mounted to the end of a hollow vertical tube that can be precisely positioned above the X ray source. The cell finding and positioning stage comprises an epi-fluorescence microscope and a feedback controlled 3 axis cell positioning stage, also mounted on the optical table. Independent vertical micro positioning of the microscope objective turret allows the focus of the microscope and the X ray focus to coincide in space (i.e. at the point where the cell should be positioned for exposure). The whole microscope stage assembly can be precisely raised or lowered, to cater for large differences in the focal length of the X ray zone plates. The facility is controlled by PC and the software provides full status and control of the source and makes use of a dual-screen for control and display during the automated cell finding and irradiation procedures.
- Research Organization:
- Gray Cancer Institute, PO Box 100, Mount Vernon Hospital, Northwood, Middlesex, HA62JR, UK
- Sponsoring Organization:
- USDOE Office of Science (SC)
- DOE Contract Number:
- FG02-01ER63236
- OSTI ID:
- 897804
- Report Number(s):
- DOE/ER/63236-1; TRN: US0703293
- Country of Publication:
- United States
- Language:
- English
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