Abstract
This work is to study the relationship between the exposure conditions and the quality of cell imaging with soft X-ray contact microscopy (SXCM). It is a crucial step in the efficient visualization of cell structures. Three different human cell lines: DU145 prostate carcinoma cells, HCC38 breast cancer cells, and Poietics mesenchymal stem cells were used to establish the optimal exposure conditions in SXCM. The image quality depended on the soft X-ray (SXR) absorbed energy and photoresist development conditions. At lower SXR energy (200 or 400 SXR pulses), sharp cell edges, membrane projections, and cell–cell connections were visible. In contrast, higher energy (600 or 800 SXR pulses) allowed observation of the cytoskeleton and the nucleus in a cell type-dependent manner (the influence of cell thickness and internal complexity was noted).
Highlights
Soft X-ray contact microscopy (SXCM) is a potent technique enabling imaging with a nanometric resolution of specific features in cells [1,2]
DU145 for imaging the different types of cell. cells, The the intracellular organellesfor were densely packed, and it was to visualize their structures experimental condition imprint preparation must bedifficult optimized for each cellinternal line separately
Soft X-ray contact microscopy may provide a powerful tool for studying cell structures
Summary
Soft X-ray contact microscopy (SXCM) is a potent technique enabling imaging with a nanometric resolution of specific features in cells [1,2]. Most structures of mammalian cells have already been visualized by electron and fluorescent microscopy, commonly used in biological research [3,4] These methods were biased by protocols for consecutive sample preparation and had insufficient resolution to analyse the structures inside the cell [5,6]. SXCM works within the energy range between the carbon and oxygen K-absorption edges known as the “water window” (2.3–4.4 nm wavelength; 284–540 eV photon energy) [9]. In this spectral region, water-containing intracellular compartments absorb much less than carbon-rich structures (e.g., plasma membranes and organelles), which absorb photons efficiently, leading to high contrast images. In the “water window”, intact biological specimens up to 10 μm thick can be observed [10,11]
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