Which solvents and chemicals are compatible with the ibidi labware?
For a quick reference, we have classified a list of solvents as either compatible or incompatible with the ibidi labware. However, some of the noncompatible solvents may still work with certain products, but only for a short time or in low concentrations. Please test the chemical compatibility of your compound with a free sample.
Lids for all ibidi products
µ-Slides, µ-Dishes, µ-Plates with ibidi Polymer Coverslip Bottom
Is there a chance of chemicals leaching into my assay when using ibidi polymer products?
No, the leaching of chemicals can be excluded, because ibidi does not use any plasticizers, stabilizers, or other chemical additives for the injection molding process of their labware products. Therefore, classic leachables, like phthalates, terephthalates, or epoxies are not present.
What are the dimensions of the ibidi labware products?
Please refer to the labware product section of our website. Here, you will find the dimensions for each specific µ-Slide, µ-Dish, µ-Plate, as well as other labware products.
Are the µ-Slides, µ-Dishes, and µ-Plates biocompatible?
All ibidi labware, including the µ-Slides and µ-Dishes, are made of biocompatible materials. The biocompatibility of all ibidi products conforms to the EN ISO 10993-5 standard (Tests for Cytotoxicity—In Vitro Methods).
What is the ibiTreat surface?
ibiTreat is a physical surface modification that provides improved cell adhesion on µ-Slides, µ-Dishes, and µ-Plates. The surface is comparable to that of standard tissue culture-treated cell culture flasks and Petri dishes. The adhesion of cells to ibiTreat µ-Slides is strong enough to perform flow experiments that simulate the physiological shear stress of blood flow. Among the ibidi products that are cited in more than 10,000 publications, the labware with an ibiTreat surface bottom is the most common.
My µ-Slides/µ-Dishes show scratches on the bottom. Where do these come from and how can I prevent them?
The ibidi Polymer Coverslip is sensitive to mechanical scratching. If you do not use extra caution when placing the ibidi µ-Slide/µ-Dish directly onto the bench or inside the incubator, it is likely to be harmed. As a result, small scratches may be visible under the microscope. To avoid this effect, we recommend protecting the bottom material by using a µ-Slide Rack or a µ-Dish Rack, for example. This solution also allows for convenient handling of up to eight µ-Slides or six µ-Dishes at once, respectively.
What is your recommendation for reusing μ-Slides, µ-Dishes, or µ-Plates?
We do not recommend reusing the slides, due to the following issues: protein remnants, maintaining sterility, cross-contamination, and more.
How can the evaporation of medium be prevented?
Depending on the incubation conditions, small volumes of medium can evaporate quickly, especially during long-term experiments. In addition, all cell culture incubators require a lengthy time to recover humidity, particularly after opening the door. While temperature and CO2 recover within minutes, full humidity recovery can take several hours. Due to these issues, we recommend using one of the following techniques for minimizing evaporation:
Place the cell culture vessel into a Petri dish that is packed with wet tissues.
Ph stands for phase contrast. The Ph+ versions of the µ-Slide 2 Well and µ-Slide 4 Well are specifically designed for phase contrast microscopy. Working with the Ph+ versions will diminish the meniscus effect, ensuring that excellent phase contrast microscopy is possible over the entire well.
The Ph+ versions are easily filled by using a standard pipette. Special openings near the well edges allow for easy access to the liquid in the well. Filling and medium exchange can be done without creating air bubbles. Due to the geometry, air bubbles cannot get trapped when the correct well volume is used. Please find a detailed description in the Instructions.
Yes. Over time, motile cells may move or proliferate over the barriers in a µ-Slide 2 Well Co-Culture, especially when they are seeded at high confluence. To avoid this, it may be helpful to coat the surface of the slide.
No, it is not possible, because there is a high chance of cross contamination between the minor wells of the µ-Slide 2 Well Co-Culture. However, two independent experiments can be performed in the two major wells.
We recommend the ibidi Mounting Medium, a glycerol-based mounting solution that is optimized for immunofluorescence stainings of, for example, adherent cells or tissue sections when using the ibidi µ-Slides, µ-Dishes, µ-Plates. The ibidi Mounting Medium is a ready-to-use solution that contains an inhibitor, which retards photobleaching of fluorochromes.
All common aqueous and non-aqueous mounting media can be used in our Immunofluorescence Chamber Slides. For mounting the glass slides, a hardening permanent mounting medium, such as Fluoroshield™ (Sigma-Aldrich), Vectashiel® (Vector Laboratories, Inc.), or ProLong® Antifade (Thermo Fisher Scientific), is recommended. Always ensure that the chosen product is suitable for your staining technique. Please note: The ibidi Mounting Medium is not recommended for immunofluorescence Chamber Slides because it is non-hardening and stays liquid.
Reusing the silicon gasket of the Immunofluorescence Chamber Slides is not recommended. Although both materials are autoclavable and compatible with alcohols, ibidi only guarantees the products’ adhesive capacity for one single use.
Due to the open well structure of the µ-Slide 18 Well - Flat and the low volume of substances being used, evaporation rates are fairly high. Therefore, the slide is only recommended for the use in short term assays that do not exceed 48 hours.
Can the sticky-Slide be detached from its substrate mounting?
All sticky-Slides make a tight seal that is not easily broken with mechanical forces. To detach the slide from its mounting, acetone can be used (e.g., by immersing the slide in a beaker of acetone). Keep in mind, however, that acetone may be incompatible with your surface or substrate.
Microscopy With the ibidi Labware
Which immersion oils are compatible with the ibidi labware products?
ibidi Glass Coverslip Bottom labware: 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. ibidi Polymer Coverslip (ibiTreat, Uncoated, and protein-coated) bottom labware: 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, and the resulting leakage may harm objectives and microscope components. All immersion oils that are not listed in the table below should be considered non-compatible.
ibidi Immersion Oil
Immersol W 2010
Silicone Immersion Oil
Which ibidi labware would you recommend for confocal microscopy?
We can recommend the ibidi Glass Coverslip bottom for all super-resolution microscopy techniques. At this time, however, only dSTORM is proven compatible with the ibidi Polymer Coverslip. We are still gathering data on the ibidi Polymer Coverslip’s compatibility with the vast number of microscopic techniques available. If you would like to take advantage of the ibidi Polymer Coverslip bottom features and test them with your own super-resolution microscopy experiments, please check out our free sample! Then, give us feedback on how the ibidi Polymer Coverslip worked with your technique of choice.
When using fluorescent microscopy, how much light can cells be exposed to during live cell imaging without being damaged?
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 cells’ behavior changes under heavy light exposure.
What are the microscopy properties of the ibidi Polymer Coverslip bottom?
The µ-Slides and µ-Dishes are optimized for inverted microscopes. The ibidi Polymer Coverslip matches the No 1.5 coverslip thickness, and is made from high-quality optical polymer material. Autofluorescence, birefringence, and the refractive index of the ibidi Polymer Coverslip are similar to those of glass, allowing for the use of all kinds of objective lenses (up to 100x oil immersion). The slide optics are designed to be used with 0.17 mm corrected objective lenses.
Is the coverslip thickness important when using inverted microscopy, and are different thicknesses available?
Yes, the thickness of the coverslip is a crucial aspect of imaging quality. Most objective lenses for microscopy are made for a coverslip thickness of 0.17 mm (170 µm, No. 1.5). A thinner or thicker substrate will require correction collars on the objective lenses to prevent the formation of blurred images due to spherical and chromatic aberrations. Therefore, in addition to the standard No. 1.5 thickness, further coverslip thicknesses are available:
0.08 - 0.12 mm
0.13 - 0.16 mm
0.16 - 0.19 mm
0.165 – 0.175 mm
0.19 - 0.23 mm
What is the thickness of the ibidi bottom materials?
The bottom materials of the ibidi µ-Slides, µ-Dishes, and µ-Plates provide a standard No. 1.5 thickness with a very low variation.
* With the 3 Well | 8 Well | 12 Well Chamber Slides, removable, ibidi provides self-adhesive, removable silicone chambers that are mounted on a standard glass slide. They are suitable for upright and inverted microscopy and enable the long-term storage of samples after immunofluorescence staining.
No, it does not occur. Unlike all other plastic materials used in cell culture, the ibidi Polymer Coverslip provides an excellent signal-to-noise-ratio, similar to glass. Please test this tremendous optical quality with a free sample.
Yes, under certain setups. The µ-Slide 18 Well - Flat was originally designed only for inverted microscopes. However, it is possible to use the slide with upright microscopes if the objectives have working distances of 2 mm or more. Alternatively, water dipping objectives, which are designed for use with upright microscopes and slides without coverslips, can be used. They can be dipped directly into the open cell culture well. Please contact your microscope supplier for these lens options.
3D Cell Culture With the ibidi Labware
How do you keep the cells in focus during a Matrigel™ assay?
Keeping the focus while taking images in time-lapse mode can be challenging. We have tested many methods, but have found that only autofocus helps.
Yes. You can, for example, embed a cell spheroid into the gel and cast it into the center well of the µ-Slide 2 Well Co-Culture. It is also possible to fill the minor wells with Matrigel and seed the cells on top. For detailed information, please refer to our Application Note, "Co-Cultivation Using ibidi Slides (AN 10) (PDF).
My collagen 3D gel looks very inhomogeneous. What is the reason for that?
Generally, an inhomogeneous appearance of the collagen gel is caused by starting polymerization too early. This can happen either due to insufficient cooling or incomplete mixing of the collagen solution. We recommend cooling the reagents you are using and keeping them on ice while preparing the 3D gel solution. We further recommend mixing the viscous collagen solution thoroughly before gel preparation.
My cells need pH 6.0 in the 3D collagen matrix. Can the gel be adjusted to a pH lower (or higher) than the 7.2–7.4 that is recommended in the instructions?
Most cell types grow best at neutral pH, which is around 7.2–7.4. However, if the experimental conditions require an environment with slightly basic or acidic pH, this can be achieved by increasing or reducing the amount of NaOH in the gel. For details, please follow the General 3D Gel Protocol (PDF), which is included in the Collagen I, rat tail Instructions.
Culture of Spheroids, Organoids, and Suspension Cells Using Bioinert
Bioinert is hydrogel-based. Does any swelling occur?
As soon as the dry surface is wetted, a swelling process occurs. This process is reversible and does not alter the passivating and optical properties.
How stable is the Bioinert surface? Will touching the Bioinert surface with a pipette tip damage it?
Touching the Bioinert surface with a pipet tip will not alter its properties. However, we recommend you avoid mechanically touching or scraping the surface.
Standard ultra-low attachments cultureware (ULA) is not suitable for fluorescence microscopy or high-resolution imaging. Is the µ-Dish 35 mm, high Bioinert suitable for these applications?
Yes, the thin Bioinert surface supports all types of high-resolution and fluorescence microscopy techniques, even with oil immersion. Bioinert is coated onto the ibidi Polymer Coverslip, which is optimized for high-resolution microscopy. The Bioinert surface itself does not interfere with the excellent optical properties of the ibidi Polymer Coverslip. The optical specifications, such as transmission, autofluorescence, refractive index, Abbe number, and birefringence, are not altered.
Single cells and spheroids do not attach to Bioinert. Does this mean that the spheroids can swim off the surface during imaging? If yes, how can this be avoided?
Even without cell attachment, the spheroids or clusters are positioned on the Bioinert surface in a stable way. When disturbing effects like strong convection, evaporation, or fast stage accelerations are not present, then there is no need to stabilize your cell samples on Bioinert. However, if a sample immobilization is necessary, we recommend increasing the viscosity of the culture medium (e.g., by using low melting agarose). We provide a detailed protocol in the Instructions (PDF).
How long is the shelf life of the Bioinert surface?
Please see the section, "Shipping & Storage" in the Instructions (PDF) for details on the shelf life.
In principle, this combination can be done to create a smaller well with the Bioinert surface. However, since the side walls of the silicone-based products are not passivated, cell adhesion is possible in that area.