µ-Slide Spheroid Perfusion

Optimal Nutrition Through Specialized Geometry. The µ-Slide Spheroid Perfusion is a specialized flow chamber for culturing free-floating 3D aggregates. It consists of 3 x 7 flat-bottomed wells, which are connected through a channel above. Each well forms its own niche, where the specimen is cultured. When applying perfusion through the channel, fresh medium continuously diffuses to the specimen. This ensures optimal nutrition and oxygen diffusion throughout the experiment without that the specimen is exposed to significant shear forces. The setup guarantees maximum of viability, but with minimum of shear stress for spheroids, organoids, or tissue.

µ-Slide Spheroid Perfusion is available with three different surfaces (Bioinert, ibiTreat, Uncoated), and is suitable for a variety of applications.

Highly Improved Spheroid Growth Rates When Cultured Under Perfusion. L929 fibroblasts show spheroid formation in the µ-Slide Spheroid Perfusion, Bioinert, days 1–14, seeding concentration 5 x 10⁵ single cells/ml. Left: no perfusion, medium exchange every second day. Right: perfusion with the ibidi Pump System, 0.75 ml/min. Phase contrast microscopy, 10x objective lens, well diameter 800 µm.

(A) Spheroid Formation of MCF-7 Breast Cancer Cells. Single cell seeding of MCF-7 breast cancer cells with spheroid formation in the µ-Slide Spheroid Perfusion, Bioinert, 1 day after seeding. Seeding concentration 5 x 10⁵ cells/ml. Phase contrast microscopy, 10x objective lens. (B) Live/Dead Staining of a Long-Term Cultured Spheroid. Live/dead FDA/PI staining of an L929 spheroid in the µ-Slide Spheroid Perfusion, Bioinert, after 14 days in culture with perfusion using the ibidi Pump System, 0.75 ml/min. Green: living cells (fluorescein diacetate, FDA); red: dead cells (propidium iodide, PI). Widefield fluorescence microscopy, 10x objective lens. (C) Fluorescence Microscopy of Adherent Cells. Fluorescence imaging of L929 fibroblasts in the µ-Slide Spheroid Perfusion, ibiTreat, fixated 2 hours after seeding. Green: F-actin (phalloidin); blue: nuclei (DAPI). Widefield fluorescence microscopy, 10x objective lens. (D) Organoid Co-Culture of PDAC Cells and Fibroblasts*. Organoid co-culture of the human pancreatic cancer (PDAC) cell line PA-TU-8988T (green, stained with CellTracker™ Green) and the murine fibroblast cell line mPSC4 (red, stained with CellTracker™ Orange CMTMR) in the µ-Slide Spheroid Perfusion. The µ-Slide was covered with a 25 µm FEP foil for matching the refractive index closer to water during upright light sheet microscopy. *The image was acquired by S. Volkery at MPI Muenster with the M Squared Aurora Airy beam upright light sheet setup. The sample was provided by K. Roth, University Marburg, Germany.