Table-top water window transmission x-ray microscopy: Review of the key issues, and conceptual design of an instrument for biology
- Departement des Micro Technologies pour la Biologie et la Sante, CEA-LETI, 17 rue des Martyrs, 38054 Grenoble (France)
As it has been beautifully demonstrated in synchrotron facilities, water window transmission x-ray microscopy (WW-TXM) has a great potential for high resolution three dimensional (3D) tomographic imaging of frozen cells, without the cumbersome staining and slicing preparation needed by electron microscopy. However, the existing instruments do not exactly meet the expectations of cell biologists in terms of performance and accessibility: 3D images of entire cells grown on a flat substrate with details in the 50-80 nm range are necessary for structural cell studies. Functional imaging is also a key issue. Specific molecular probes are widely used to achieve molecular imaging in optical and electron microscopy. The same demand applies to x-ray microscopy. Immediate availability of the observation technique within the biology laboratory is as important as its performance. Therefore, WW-TXM will develop on a wider scale only when table-top instruments become available. We present a detailed analysis of such a microscope. The source is clearly the key element. Laser created plasmas of carbon or nitrogen are a proven but expensive solution. Cerenkov emission in vanadium has been demonstrated as a potential monochromatic source, but we emphasize severe obstacles: huge thermal load and radiation protection. We show that oxygen K{alpha} line excitation by an electron beam is a realistic alternative. Being a purely monochromatic source, it would allow the use of a high efficiency mirror condenser, while laser plasmas imply spectral selection with the associated losses. We then describe the main elements of an affordable laboratory microscope, supported by numerical simulations and preliminary experimental work. We also show that functionalized quantum dots, currently used in fluorescence microscopy, are equally detectable with soft x-rays and would allow a dual modality observation. Finally, the expected performance of this prototype is discussed and confronted by the requirements of cell biology.
- OSTI ID:
- 20723112
- Journal Information:
- Review of Scientific Instruments, Vol. 76, Issue 9; Other Information: DOI: 10.1063/1.2018633; (c) 2005 American Institute of Physics; Country of input: International Atomic Energy Agency (IAEA); ISSN 0034-6748
- Country of Publication:
- United States
- Language:
- English
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