Imaging XPS and photoemission electron microscopy; surface chemical mapping and blood cell visualization.

ABSTRACT

Combined photoemission electron microscopy (PEEM) and imaging x-ray photoelectron spectroscopy (XPS), i.e., electron spectroscopy for chemical analysis in the nanoregion, has been used for surface characterization of bio-relevant and biological samples. In the first example, the authors prepared a gold patterned silicon substrate, stepwise surface modified by self-assembled monolayers followed by quantum dot (QDot) specific linking and investigated by means of work function mapping and elemental imaging in the submicrometer range. Spatially resolved core level images of C1s, V2p, and Y3d are obtained, which verify the selective thiol adsorption on the gold squares and specific binding of europium doped yttrium vanadate QDots on the self-assembled monolayer. The second example is platelet adhesion to Immunoglobulin G modified silicon surfaces, investigated by means of laterally resolved PEEM. Images of platelets clearly show activated cells with a morphology change including an enlarged surface area and elongated pseudopodia, with a lateral resolution of 140 nm. In the last example, neutrophils were allowed to attach to plain silicon surfaces and investigated by means of PEEM and imaging XPS. Here, the cells show a round shaped morphology, as expected. Threshold imaging with work function contrast is used to localize the area of interest, followed by elemental specific mapping on cells in the submicrometer region. Chemical shifts of C1s in photoemission are used to distinguish vital parts of the cell structure. The strong C1s (C-C) signal is achieved from the region of the cell membrane, i.e., high density of phospholipids, while C1s (C-N) and C1s (C-O) signals are obtained from the core of the cell, in good agreement with the presence of cytoplasm and deoxyribonucleic acid containing cell nucleus. The combination of PEEM and imaging XPS is shown here as a tool to deliver new insight into biological samples, i.e., a rapid sample overview is obtained based on low energy secondary electrons with work function contrast, followed by detailed studies in the narrow mode for elemental compositions based on photoemission. This study illustrates the strength of combined PEEM and XPS in the imaging mode on cell studies.