Cryo-soft X-ray tomography is an imaging technique that addresses the need for mesoscale imaging of cellular ultrastructure of relatively thick samples without the need for staining or chemical modification. It allows the imaging of cellular ultrastructure to a resolution of 25–40 nm and can be used in correlation with other imaging modalities, such as electron tomography and fluorescence microscopy, to further enhance the information content derived from biological samples. An overview of the technique, discussion of sample suitability and information about sample preparation, data collection and data analysis is presented here. Recent developments and future outlook are also discussed.
Poniższy raport przedstawia sposób zarządzania wartością przedsiębiorstw z branży wyrobów drewnianych z uwzględnieniem jednego z czynników kreującego wartość firmy, jakim jest kapitał obrotowy netto.
Hard X-ray fluorescence microscopy and magnified phase contrast imaging are combined to obtain quantitative maps of the projected zinc mass fraction in whole cell of PC12 cell lines. The experiments were performed on freeze dried cells at the nano-imaging station ID22NI of the European Synchrotron Radiation Facility (ESRF). X-ray fluorescence analysis gives the areal mass of most major, minor and trace elements while quantitative phase contrast imaging provides maps of the projected mass. The combined method was validated on calibration samples by comparison with other alternative techniques such as Atomic Force Microscopy (AFM) and Scanning Transmission Ion Microscopy (STIM). Up to now, absolute quantification at the sub-cellular level was impossible using X-ray fluorescence microscopy but can be reached for the first time with the use of the proposed approach.
Hard X-ray fluorescence microscopy and magnified phase contrast imaging are combined to obtain quantitative maps of the projected metal concentration in whole cells. The experiments were performed on freeze dried cells at the nano-imaging station ID22NI of the European Synchrotron Radiation Facility (ESRF). X-ray fluorescence analysis gives the areal mass of most major, minor and trace elements; it is validated using a biological standard of known composition. Quantitative phase contrast imaging provides maps of the projected mass and is validated using calibration samples and through comparison with Atomic Force Microscopy and Scanning Transmission Ion Microscopy. Up to now, absolute quantification at the sub-cellular level was impossible using X-ray fluorescence microscopy but can be reached with the use of the proposed approach.
Hard X-ray fluorescence microscopy and magnified phase contrast imaging are combined to study radiation effects on cells. Experiments were performed on freeze-dried cells at the nano-imaging station ID22NI of the European synchrotron radiation facility. Quantitative phase contrast imaging provides maps of the projected mass and is used to evaluate the structural changes due to irradiation during X-ray fluorescence experiments. Complementary to phase contrast imaging, scanning transmission ion microscopy is performed and doses of all the experiments are compared. We demonstrate the sensitivity of the proposed approach to study radiation-induced damage at the sub-cellular level.
The ESRF synchrotron beamline ID22, dedicated to hard X-ray microanalysis and consisting of the combination of X-ray fluorescence, X-ray absorption spectroscopy, diffraction and 2D/3D X-ray imaging techniques, is one of the most versatile instruments in hard X-ray microscopy science. This paper describes the present beamline characteristics, recent technical developments, as well as a few scientific examples from recent years of the beamline operation. The upgrade plans to adapt the beamline to the growing needs of the user community are briefly discussed.