Controlled Cell Adhesion With ibidi Micropatterning
The ibidi µ-Patterning technology enables spatially defined cell adhesion for various 2D and 3D cell culture applications.
Miniaturized adhesive patterns (e.g., lines, squares, or dots) are irreversibly printed on the non-adhesive Bioinert surface of the ibidi Polymer Coverslip, allowing for precisely controlled cell adhesion. The µ-Patterns are dry-stable, sterile, and ready to use.
For various single cell assays (e.g., CAR-T cell activity assay)
For culturing 3D cell aggregates (e.g., spheroids or organoids) or cell patch monolayers
For determining the most suitable micropatterning geometry for the respective application
- Long-term stable, and biologically inert surface
- Superior to the standard ultra-low attachment (ULA) surfaces: no cell or protein adhesion (full passivation)
- Layered onto the ibidi Polymer Coverslip—the highest optical quality for imaging
Learn more here.
|Size (resolution)||> 3 µm; different sizes possible|
|Geometry||Circles, squares, lines, your specific geometry|
|Available surfaces||- Specific cell adhesion (RGD or specific molecule/peptide)
- unspecific cell and molecule adhesion
- Custom-specific adhesion via click chemistry
|Optics||- Very low autofluorescence for high resolution imaging
- No visibility of µ-Patterns in phase contrast/brightfield
- Optional µ-Pattern visibility under fluorescence
The size of the µ-Pattern can be adapted to the morphology of the cell type of interest, so that an array of single cells can be conveniently analyzed using applications such as high-resolution imaging.
Single-cell array with RCC26 tumor cells. µ-Patterning was done with RGD adhesion spots on the Bioinert surface. Spot size 40 µm x 40 µm. Phase contrast microscopy, 10x objective lens.
By using different geometries and sizes of the µ-Patterning, multi-cell arrays can be performed with defined adhesion for various applications, such as high-resolution imaging.
Multi-cell array with Rat1 cells. µ-Patterning was done with RGD adhesion spots on the Bioinert surface. Spot size 200 µm x 200 µm. Phase contrast microscopy, 4x objective lens.
Defined adhesion spots, surrounded by Bioinert, are able to catch all adherent single cells from a cell suspension. Bioinert is fully non-cell-attachable. This forces all cells to aggregate to each other at the adhesion spots, thus forming spheroids in a defined and controllable way.
Suspension of NIH-3T3 cell line seeded on 200 µm adhesion spots, 64 hours live cell imaging, phase contrast, 4x objective lens.
Spheroid and Organoid Culture
The size and the cell adhesion properties of the µ-Patterning can be adjusted to hold 3D spheroids and microtissues in position. Using this setup, cells in 3D can be studied during proliferation, differentiation, invasion, and migration.
NIH-3T3 3D spheroids (left) plated on adhesive, Cy3-labeled spots (right) with a 300 µm diameter. 10x objective lens.
Generation of 3D spheroids from HT-1080, MCF-7, and NIH-3T3 cells. The spheroids were generated using the agarose assay, then put on 300 µm adhesive spots for positioning. Phase contrast, 4x objective lens.
Immunofluorescence staining of spheroids from RCC26 (left) and Rat1 cells (right) in in the µ-Slide VI 0.4 Luer, patterned with 200 µm circles at 600 µm distance. The cells were stained with phalloidin (green) and alpha-tubulin (red). Nuclei were stained with DAPI (blue). Widefield fluorescence microscopy, 4x and 10x objective.
Spheroid Culture Under Constant Media Circulation
By creating micropatterns within a µ-Slide VI 0.4 Luer, aggregates on the pattern can be constantly supplied with fresh media by perfusing the system with the ibidi Pump System.
Fibroblasts (3T3) under flow conditions at 3 dyn/cm2 compared to the static control, cultured in the µ-Slide VI 0.4 Luer for 14 days.
By applying a constant media flux around the cell aggregates, spheroids become more compact and round.
Fibroblast (3T3) aggregates under flow conditions at 3 dyn/cm2 form a compact and round spheroid over time.
3T3 fibroblasts were seeded on a patterned µ-Slide VI 0.4. A shear stress of 3 dyn/cm2 was applied 7 days after cell seeding. 15 h time lapse microscopy, 4x objective.
CAR-T Cell Killing Assays
CAR-T cells represent a promising new cancer therapy tool. Live cell imaging allows to analyze T cell/cancer cell interaction in real time with single cell resolution. However, analysis of confluent cell layers is very time-consuming and therefore not possible in high throughput screens. To facilitate high throughput label-free analysis of T cell potency in a live cell imaging setup, we generated arrays of homogenously distributed cancer cells. By combining optical analysis and advanced image processing, cytotoxic T cell activity over time on a single cell level can be evaluated without the use of any labeling.
Single Cells in a 2D Environment
Time lapse microscopy of a CAR-T cell killing assay with RCC-26 tumor cells and JB4 T cells on a single cell pattern. Data were analyzed using FastTrack AI by MetaVi Labs.
Multicell Spots in a 3D Collagen Matrix
RCC-26 cancer cells immobilized on multi cell pads. Effector T cells applied in a collagen I matrix (Collagen Type I, Rat Tail) induce apoptotic body formation of cancer cells.
Presented at the Annual Meeting of the Biophysical Society 2020, San Diego, USA.
Presented at the ASCB|EMBO Meeting 2019, Washington DC, USA.
Presented at the µTAS Conference 2019, Basel, Switzerland.