Correlative imaging allows the investigation of biological samples under various aspects. We are working towards a correlated extreme ultraviolet (EUV), infrared (IR) and visible (VIS) fluorescence microscope in a single integrated setup. With ptychography, which is a lensless imaging technique, quantitative amplitude and phase information are obtained. EUV ptychography provides resolution in the nanometer scale and an excellent element contrast, but suffers from low throughput due to limited coherent photon flux. Using longer wavelengths such as UV/visible and Near-IR enables high-speed imaging with sub micrometer resolution. Further, the combination with fluorescence detection adds functional contrast with micrometer-scale resolution. Here, we demonstrate in a proof-of-concept experiment correlated ptychography-fluorescence microscopy in the visible range. By using the reconstructed beam from the ptychography measurement the fluorescence scanning map can be deconvolved, which significantly improves the resolution.
EUV ptychography combines high resolution, strong material contrast, reasonable penetration depth, and easy sample preparation. It thus has the potential to bridge the gap between visible light and electron microscopy. In this contribution, we present recent results on table-top ptychographic EUV microscopy. The experimental setup relies on a high photon flux 13.5 nm wavelength high-order harmonic source. A simple amplitude mask upstream of the sample tailors the illumination. The sample is scanned by piezo-driven stages. Finally, far-field diffraction patterns are recorded by an EUV detector and fed into a ptychographic iterative engine to retrieve both the illumination and the sample transmission in amplitude and phase. The resolution of the EUV microscope (16 nm) was characterized using a Siemens star test sample. In the next step, a thin lamella of an integrated circuit was investigated. The resulting EUV images exhibit a rich diversity in amplitude and phase. We analyzed that the relative amplitude precision is better than 4% and the absolute phase precision as good as ~ 20 mrad. In this way, the microscope provides excellent input data for further analysis of the material composition. For this purpose, the projected scattering quotient is calculated from the reconstructed phase and amplitude and compared to tabulated material parameters. Here, materials like Al, Si3N4, and SiO2 were determined with high sensitivity. Finally, biological samples, namely germlings of the filamentous fungus Aspergillus nidulans were investigated. A similar scattering quotient analysis allowed us to clearly distinguish regions with low- and high lipid and phospholipid concentrations.
In this contribution, we present a scanning coherent diffractive imaging (i.e. ptychography) microscope operating in the EUV. Coherent EUV radiation at 13.5 nm is generated by high-order harmonic generation using a high-power fiber laser system. Utilizing structured illumination, a highly stable EUV source and ptychography setup sub 20 nm half-pitch resolution is demonstrated on a resolution test chart. Moreover, the lamella of an integrated structure is investigated and its contained materials are identified using the measured quantitative amplitude and phase.
In recent years coherent diffraction imaging (CDI) has evolved into a mature technology. Thanks to its lensless nature, it allowed to bypass the limitations of X-ray optics. At the same time, laser development in combination with high harmonic generation (HHG) has pushed the coherent XUV photon flux to values comparable to 3rd generation synchrotron facilities, which enables lensless imaging experiments that were previously only possible at large-scale facilities. Furthermore, the intrinsic short pulse duration of HHG radiation has potential for imaging experiments down to attosecond time scales. In this contribution, we present our latest results on lensless imaging using a fiber laser driven HHG source at 92 eV. A high photon flux source is used for scanning coherent diffractive imaging (ptychography) demonstrating sub-50 nm resolution. Further, an extension to Fourier transform holography is shown, which enables to increase the useable bandwidth by a factor of five without sacrificing spatial resolution. This paves the way for combing high-resolution table-top lensless imaging with attosecond pump-probe experiments.
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