FIB Preparation and SEM Investigations for Three‐Dimensional Analysis of Cell Cultures on Microneedle Arrays

Abstract

We report the investigation of the interfaces between microneedle arrays and cell cultures in patch-on-chip systems by using Focused Ion Beam (FIB) preparation and Scanning Electron Microscopy (SEM). First, FIB preparations of micro chips are made to determine the size and shape of the designed microneedles. In this essay, we investigate the cell-substrate interaction, especially the cell adhesion, and the microneedle's potential cell penetration. For this purpose, cross-sectional preparation of these hard/soft hybrid structures is performed by the FIB technology. By applying the FIB technology followed by high-resolution imaging with SEM, new insights into the cell-substrate interface can be received. One can clearly distinguish between cells that are only in contact with microneedles and cells that are penetrated by microneedles. A stack of slice images is collected by the application of the slice-and-view setup during FIB preparation and is used for three-dimensional reconstruction of cells and micro-needles.

3D imaging of cell interactions with electrospun PLGA nanofiber membranes for bone regeneration

“…FIB-SEM tomography is widely used in studying microstructure changes in superalloys [40] as well as biological and geological materials [41]. Recently, FIB-SEM has been used to investigate the cell-substrate interaction between microneedle arrays [42] and complex structures including interconnections of dentine tubules [39]. Despite the introduction of FIB-SEM as a high-resolution imaging tool, evaluation of the cellular interactions with porous tissue scaffolds is lacking.…”

Role of α2β1 integrins in mediating cell shape on microtextured titanium surfaces

“…FIB milling has traditionally been used to prepare various materials for transmission electron microscope analyses . It can also be used to prepare cross sections of biological samples for interrogating cell/surface interfaces . We systematically investigated changes in cell morphology with varying microroughness using FIB/SEM, cytomorphometric analysis, and three‐dimensional (3D) reconstructions.…”

“…25,26 It can also be used to prepare cross sections of biological samples for interrogating cell/surface interfaces. [27][28][29][30][31] We systematically investigated changes in cell morphology with varying microroughness using FIB/ SEM, cytomorphometric analysis, and three-dimensional (3D) reconstructions. Cells were cultured on smooth and microtextured Ti surfaces for 3 days, during which initial attachment occurred and mRNA could be generated in response to surfaces.…”

PoreGenic® - Patch-on-Chip-System für Zellnetzwerke

“…Allerdings ist eine zusätzliche Interpretation der aufgezeichneten Potentiale nicht möglich, da vor allem die Chiptypen A-C mit Silizium-Substrat starke photoelektrische Effekte durch die Beleuchtung des Mikroskops zeigen und damit die Auswertung verfälschen würden. Zum qualitativen Nachweis der Zellpenetration wird die Rasterelektronenmikroskopie (REM) zusammen mit einer fokussierten Ionenstrahlpräparation (FIB) kombiniert [12,13]. Die resultierenden Querschnittsdarstellungen sollen den Erfolg der lokalen Elektroporation an der Schnittstelle von Nadelelektrode und Zelle dokumentieren, siehe Bild 2 D und Bild 3 A&B. Dazu werden die Zellen sofort nach den Versuchen fixiert, dehydriert und mit einer dünnen leitenden Platinschicht überzogen.…”