Photoemission electron microscopy and imaging X-ray photoelectron spectroscopy are today frequently used to obtain chemical and electronic states, chemical shifts, work function profiles within the fields of surface- and material sciences. Lately, because of recent technological advances, these tools have also been valuable within life sciences. In this study, we have investigated the power of photoemission electron microscopy and imaging X-ray photoelectron spectroscopy for visualization of human neutrophil granulocytes. These cells, commonly called neutrophils, are essential for our innate immune system. We hereby investigate the structure and morphology of neutrophils when adhered to gold and silicon surfaces. Energy-filtered imaging of single cells are acquired. The characteristic polymorphonuclear cellular nuclei divided into 2-S lobes is visualized. Element-specific imaging is achieved based on O 1s, P 2p, C 1s, Si 2p, and N is core level spectra, delivering elemental distribution with submicrometer resolution, illustrating the strength of this type of cellular morphological studies.

Photoemission electron microscopy (PEEM) and imaging X-ray photoelectron spectroscopy (XPS) have over the years been powerful tools in classical surface physics and material sciences, and due to recent technological advances, their uses within other fields/disciplines are rapidly growing. Lately, the XPS/PEEM based elemental analysis and characterization in imaging mode, with exquisite spatial resolution and high sensitivity, has shown the potential to deliver new mechanistic insights in cell-biology/medicine. In this work, the aim was to visualize biological processes on the cellular level, with the additional dimension of topographical morphology and element specific information, mapping chemical composition and chemical states. This is hereby demonstrated by combined PEEM and imaging XPS investigation of neutrophils and their activation processes, where fluorescence microscopy commonly used in biology is used for benchmarking. Neutrophils are phagocytic cells and are vital components in the human immune system, with the fundamental role of fighting invading pathogens. They are capable of ingesting microorganisms or particles, and in order to capture and trap foreign objects, one of their strategies is to release nuclear DNA by the formation of extracellular web-like traps (NETs). Here, we report how neutrophils are triggered by controlled nanoparticle (NP) exposure. The neutrophils and NETs formation are imaged in the presence of NPs, and we report the elemental composition of single cells and the structure of NETs. Cellular uptake of nanoparticles is proven and the states just before and after NETs release are imaged, as well as visualization of the extraordinary capability for mass transport at distances 10 times or more than the size of the cell itself. This method paves the way for element specific imaging of biorelated cells on surfaces as well as nanoparticle tracking in the submicro- and nanoregions.

Neutrophil granulocytes are the most abundant white blood cells in mammals and vital components of the immune system. They are involved in the early phase of inflammation and in generation of reactive oxygen species. These rapid cell-signaling communicative processes are performed in the time frame of minutes. In this work, the activity and the response of neutrophil granulocytes are monitored when triggered by cerium-oxide based nanoparticles, using capacitive sensors based on Lab-on-a-chip technology. The chip is designed to monitor activation processes of cells during nanoparticle exposure, which is for the first time recorded on-line as alteration of the capacitance. The complementary metal oxide semiconductor engineering chip design is combined with low temperature co-fired ceramic, LTCC, packaging technology. The method is label free and gently measures cells on top of an insulating surface in a weak electromagnetic field, as compared to commonly used four-point probes and impedance spectroscopy electric measurements where electrodes are in direct contact with the cells. In summary, this label free method is used to measure oxidative stress of neutrophil granulocytes in real time, minute by minute and visualize the difference in moderate and high cellular workload during exposure of external triggers. It clearly shows the capability of this method to detect cell response during exposure of external triggers. In this way, an informationally dense non-invasive method is obtained, to monitor cells at work.