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FAQ: Light Microscopy

What is Differential Interference Contrast (DIC) Microscopy?

DIC uses polarized light to convert phase delays into intensity changes (contrast). The effect is called differential, because contrast is created only in neighboring areas.

Which kind of µ-Slides would you recommend for confocal microscopy?

For confocal microscopy, all ibidi µ-Slides and µ-Dishes work well. The general recommendation is to use a glass coverslip, or a coverslip-like plastic material, such as the ibidi Polymer Coverslip products.

Are ibidi materials compatible with super-resolution microscopy (STORM, PALM, STED)?

Super-resolution is possible with the ibidi Polymer Coverslip, but we recommend using a glass bottom.

What is the Halo effect?

The halo effect is an unfortunate artifact in phase contrast microscopy. It is the display of spurious bright areas around phase objects, or the reverse contrast in images. The effect is seen very often with specimens that induce large phase shifts, such as round cells during mitosis.

Is there an existing chamber where you can mount your own pre-treated circular coverslips?

Not as far as we know, but one option is to use the ibidi sticky-Slides (for rectangular coverslips 25 mm x 75 mm). The ibidi µ-Dish 35 mm, high Glass Bottom can also work, but it is already pre-assembled for direct use.

When using a microscope with a 40x objective and a 2x zoom, will it be strong enough to show intracellular events?

Yes, you will most certainly see intracellular events. With a demo experiment, you can test whether or not the image quality is good enough. Please contact your microscope/objective lens supplier and ask for a demo system.

How do you keep the cells in focus for a Matrigel™ assay?

Keeping the focus of the low-resolution movie (e.g., the tube formation assay) can be challenging. We have tested many methods, but have found that only autofocus helps. This issue is also the cause of the illumination changes, because the system automatically adjusts the focus, brightness, and contrast.

What are the criteria on how much light the cells can get, without being damaged during live time series in confocal fluorescent microscopy?

The general rule is that the least amount of light possible is best for the cells. Additionally, we recommend doing control experiments with a very low light exposure (which will result in a bad image quality), just to test whether or not the cell’s behavior changes under heavy light exposure.

Which immersion oils are compatible with ibidi products?

For ibidi Glass Bottom products:

When using ibidi Glass Bottom products with oil immersion objectives, there is no known incompatibility with any immersion oil on the market. All types of immersion oils can be used.

For ibidi Polymer Coverslip (ibiTreat, uncoated, and protein-coated):

When using oil immersion objectives with the ibidi Polymer Coverslip, use only the immersion oils specified in the table below. The use of any non–recommended oil could damage the ibidi Polymer Coverslip. The resulting leakage may harm objectives and microscope components. All immersion oils that are not listed in the table below should be considered as non-compatible.

Company Product Ordering Number Lot Number Test Date
ibidi ibidi Immersion Oil 50101 16-12-27 01/2017
Zeiss Immersol 518F 444960 160706 01/2017
Zeiss Immersol W 2010 444969 101122 04/2012
Leica Immersion Liquid 11513859 n.a. 03/2011
Cargille Type A 16482 100592 01/2017
Cargille Type HF 16245 92192 01/2017
Olympus Silicone Immersion Oil SIL300CS-30CC N4190800 01/2017
Carl Roth Immersion oil X899.1 414220338 01/2017