Product information µ-Slide Angiogenesis The µ-Slide Angiogenesis is a cell culture product for angiogenesis assays or high resolution cell microscopy. Cells can be grown on a gel-matrix, e.g. BD MatrigelTM (Becton-Dickinson) or directly on the ibidi coverslip-like plastic bottom. © ibidi GmbH, Version 1.0, January 24th, Elias Horn

What is angiogenesis? Definition: Formation of new blood vessels to tumors Aim: Hinder the new vessels and you hinder tumor growth! Assays: Tube formation assay Sprouting assay Angiogenesis is an interesting approach in biomedical research. The word angiogenesis means “formation of blood vessels” When tumor cells grow they develop a huge demand for oxygen and nutrients. New blood vessels are needed and progenitor endothelial cells start to grow into the tumor. Without this tumor feeding, tumor cell growth is assumed to be limited. A better understanding of this process is a key feature in future cancer therapies. © ibidi GmbH

In vitro angiogenesis assays I Tube formation assay: Cells cultivated on a gel matrix form tubes and nodes. In a typical tube formation assay single cells (e.g. endothelial cells) are seeded on a gel matrix. Several types of collagen gels and BD MatrigelTM (Becton-Dickinson) are commonly used. After a while the cells rearrange to specific net-like patterns. © ibidi GmbH

In vitro angiogenesis assays II Sprouting assay: Cell spheroids or aortic rings cultivated on a gel matrix form sprouts. In a typical sprouting assay, a cell spheroid or piece of tissue (aortic ring) is placed onto a gel matrix. After a cell-specific time the endothelial cells form sprouts. © ibidi GmbH

Features I µ-Slide Angiogenesis 15 wells with numbers and letters 4 mm wells in 5 mm wells Inner well for 10 µl gel matrix Upper well for 50 µl medium Standard slide format Lid for low evaporation Compatible with multi-channel pipettes The µ-Slide Angiogenesis is a cell culture system based on a well-in-well technique. A small 10 µl inner well is designed for gel matrices (e.g. BD MatrigelTM, collagen gel, agarose). After the gel is polymerized 50 µl cell suspension can be filled into the upper well. This makes the cells grow on a defined gel matrix. The area between the wells can be filled with distilled water to further decrease evaporation effects. This is very useful for long-term experiments. For direct cell culture and unrestricted microscopy the µ-Slide Angiogenesis can also be used without gel matrices. The well-to-well distance of 9 mm is compatible with multichannel pipettes as used for 96 well plates. © ibidi GmbH

Features II Excellent optical properties for microscopy Homogeneous cell growth Compatible with staining and fixation Biocompatible plastic material – no glue, no leaking For various types of gels (MatrigelTM, collagen, agarose)* When the µ-Slide Angiogenesis is filled properly with 10 µl gel matrix and 50 µl cell medium there is no meniscus formation. The two plane surfaces provide good phase contrast and very homogeneous cell distribution. *Please note: The gel matrix is not part of the product! © ibidi GmbH

Filling and handling Short protocol for µ-Slide Angiogenesis Recommended protocol for µ-Slide Angiogenesis: Fill 10 µl liquid gel into the inner wells and close the slide using the lid. Polymerize the gel matrix following the manufacturer’s instructions. Seed cells in 50 µl cell suspension on top. Conduct your experiment. Gel pipetting Gel polymerization Cell seeding Experiment © ibidi GmbH

No meniscus formation in µ-Slide Angiogenesis Optical improvement I No meniscus formation in µ-Slide Angiogenesis All cells in focus Meniscus in 96 well plate Only few cells in focus In µ-Slide Angiogenesis there is almost no meniscus formation at the air-water interphase and the water-gel interphase. Due to the fact that all surfaces are parallel (not bent) to the optical plane there is only few image distortion and very homogeneous cell distribution. Depth of focus © ibidi GmbH

Optical improvement II No meniscus formation in µ-Slide Angiogenesis All cells in focus Meniscus in 96 well plate Only few cells in focus The parallel beam path using µ-Slide Angiogenesis allows growing all cells in one focal plane on the gel surface. This supports microscopy of large fields of view (with 5x and 10x objective lenses). Source: Peter Nelson, LMU, Munich, Germany © ibidi GmbH

Angiogenesis assays in µ-Slide Angiogenesis vs. 96 well plate 10 µl gel per well* No meniscus Excellent optics 15 wells Slide format 96 well plate 100 µl gel per well* Meniscus formation Poor optics 96 wells Multi well format There are two major improvements when using µ-Slide Angiogenesis instead of a common 96 well plate for angiogenesis assays. Only 10 µl gel per well/assay. (3D matrices are very expensive.) Better optical quality. (No meniscus.) *Saves 90% gel per assay. In case of BD MatrigelTM the costs are reduced from approx. 3€ to 0.3€ per well/assay. © ibidi GmbH

Microscopy properties I Cells on gel matrix Cells in gel matrix Cells without gel matrix Only for objective lenses with working distances over 1 mm For all objective lenses (up to 100x) and all microscopy techniques Due to the gel there is only limited access to high resolution microscopy when the cells grow on top of the gel matrix surface. Cells embedded into the gel can be observed with less restrictions. The µ-Slide Angiogenesis can also be used without any gel matrix. This is an interesting approach for stem cell assays with only few cells. Without the gel µ-Slide Angiogenesis is suitable for all kinds of microscopy techniques based on transmitted light and fluorescence. Even DIC is possible without using the lid. © ibidi GmbH

Microscopy properties II 10 µl gel create a plane surface ~ 5 µl gel create a bent surface When 10 µl gel is used there is a plane surface providing good optical properties and homogeneous cell distribution. Using only approx. 5 µl gel matrix creates a bent gel surface. This can be useful for small amounts of cells (stem cells) which need to be accumulated in the center. Good optics Homogeneous cell distribution Mini-well for few cells Cell accumulation in the center © ibidi GmbH

Applications and assays Cells on or in gels (e.g. BD MatrigelTM) Tube formation Sprouting assays Aortic ring assay Cell spheroids General cell culture Numerous assays and experiments require cells on or in gel matrices. Source: Peter Nelson, LMU, Munich, Germany © ibidi GmbH

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