1. 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

2. 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.
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3. 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.
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4. 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.
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5. 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.
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6. 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!
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7. 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
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8. No meniscus formation in µ-Slide Angiogenesis
Optical improvement I
No meniscus formation in µ-Slide Angiogenesis
All cells in focus
Meniscus in 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
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9. Optical improvement II
No meniscus formation in µ-Slide Angiogenesis
All cells in focus
Meniscus in 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
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10. 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.
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11. 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

12. 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
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13. 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

14. Free sample program Get free samples on:
ibidi will send free samples worldwide to all customers and distribution partners. Please visit to fill the free sample request form.
ibidi's free sample program is free of charge.
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