FAQ

Common questions on µ-Slides

Design & Properties

Physical & Chemical Properties

Can you tell me the dimensions of the Slides?
How are the microscopy properties of the slide/dish bottom
(thickness, autofluorescence, birefringence, refractive index...)?
What is the optical quality compared to glass?
Which solvents are compatible with the µ-Slides?
What is the shear stress inside a µ-Slide?

Biological Properties

Are the slides biocompatible?
What is the ibiTreat surface?
Which surface coatings are offered by ibidi?
Which cell lines have been tested so far?
How are the cells supplied with oxygen and CO₂?

Handling

How to fill the µ-Slide I properly?
How to remove and avoid air bubbles in channels?
What is the optimal mounting solution used in immunofluorescence?
Can I use the Slides for suspension cultures?
What can I do against evaporation?

Design & Properties

Physical & Chemical Properties

Can you tell me the dimensions of the Slides?
Please have a look in the µ-Slide section on our website. There all the dimensions are given for all Slides.

How are the microscopy properties of the slide/dish bottom
(thickness, autofluorescence, birefringence, refractive index...)?
The µ-Slides and µ-Dishes are optimized for inverted microscopes. The bottom matches No 1.5 coverslip thickness and is made from high quality plastic material. Autofluorescence, birefringence, and refractive index of the bottom are similar to those of glass allowing the use of all kinds of objective lenses (1x up to 100x oil immersion). Our slide optics is designed to be used with 0.17 mm corrected objective lenses.

Thickness of coverslips:

(Source: Gerhard Menzel, Glasbearbeitungswerk GmbH & Co. KG, Braunschweig, Germany)

What is the optical quality compared to glass?
Our polymer has almost the same optical quality as glass. In the PDF we show a comparison between the autofluorescence signal of the CCD camera noise, a µ-Slide and a coverslip. Additionally some normally used plastic materials like petri dishes are compared.
PDF (236 KB)

Biological Properties

Are the slides biocompatible?
The slides and dishes are made of biocompatible materials. The biocompatibility of all types of ibidi-slides and ibidi-dishes is tested regarding EN ISO 10993-5 (Test for cytotoxicity: in vitro methods)

What is the ibiTreat surface?
ibiTreat is a physical surface modification for improved cell adhesion on µ-Slides. The surface is comparable to standard cell culture flasks and Petri dishes. The adhesion of cells to ibiTreat µ-Slides is strong enough to perform flow experiments simulating the physiological shear stress of the blood flow. The surface was tested with over 20 different cell lines and primary cells.

Which surface coatings are offered by ibidi?
The growth, development, and signaling of cells cultured on surfaces depend strongly on the adhesion factors and surface modifications your cell culture labware is coated / treated with. One of the advantages of the ibidi µ-Slides is, that these can be treated similarly to standard plastic labware.
For standard cell culture applications ibidi offers µ-Slides with the following approved surfaces.

coating procedure PDF (37 KB)

Tissue culture treated surface – ibiTreat
ibiTreat is a physical surface modification for improved cell adhesion on µ-Slides. The surface is comparable to standard cell culture flasks and Petri dishes. The adhesion of cells to ibiTreat µ-Slides is strong enough to perform flow experiments simulating the physiological shear stress of the blood flow. The surface was tested with over 20 different cell lines and primary cells.

Collagen IV
Collagen IV is the primary collagen found in large, extracellular basement membrane structures and complex organs. Collagen IV substrates have been tested for a variety of standard cell lines, epithelial, endothelial, nerve, and muscle cells. The µ-Slides are coated with a mouse Collagen IV.

Poly-L-Lysine (PLL) / Poly-D-Lysine (PDL)
PLL / PDL is a polymer of the amino acid L / D-Lysine. It is one of the most commonly used adhesion substrates suitable for a large variety of cell types. It has been used especially for neuronal cultures. Adhesion via this polymer is mediated by an integrin-independent mechanism.

Fibronectin
Fibronectin is a glycoprotein widely used in cell culture coatings. It plays an important role in cell surface interactions, which is mediated by an RGD motive. It allows neurite out- growth and has been used for glial and neural cells.
Due to its short shelf life, Fibronectin coated µ-Slides are available on demand only.
Delivery time is about three weeks.

Hydrophobic, uncoated
This surface permits no direct cell growth and can be used for own specific coating procedures or for non adherent cells.

Which cell lines have been tested so far?
We have been working with the following cell lines internally:
HBE; HELA; RAT1, 3T3, NRK´s, HUVEC, CHO, COS7, 293, PC-12, B16, HCV29

If you want to know about growth of other cell lines in ibidi µ-Slides, please do not hesitate to contact us.

How are the cells supplied with oxygen and CO₂?
Like most conmon plastics, the used plastic has a higher permeability for O₂ and CO₂ than glass. Also in the media, a lot of O₂ is dissolved. Because of the height of the channel (µ-Slide I, µ Slide VI, µ-Slide y-shaped), enough O₂ and CO₂ is available for a long term experiments.

Handling

How to fill the µ-Slide I properly?
Please have a look at the videos showing the proper handling of the µ-Slides.
movie (1667 KB)

How to remove and avoid air bubbles in channels?
If you culture the cells over a longer time some air bubbles can occur inside the channel. Trapped bubbles can be removed from the channel by inclining the µ-Slide and carefully knocking at one edge.
To avoid emerging bubbles, put your media and the µ-Slide in a non gas-tight container and put both into your incubator the day before use. This will lead to equilibration of dissolved gasses.

What is the optimal mounting solution used in immunofluorescence?
We recommend to use 80% glycerol in PBS with added anti-bleaching agents like DABCO (2%), paraphenylendiamine, or n-propylgallate. The slides can be stored for approx. 4 weeks.

Can I use the Slides for suspension cultures?
Currently, we are working on a µ-Slide with a filter at one edge allowing to use the µ Slides in suspension cultures. This is still in a prototyping stage. Please contact us for further details.

What can I do against evaporation?
Depending on the incubating conditions, small volumes might evaporate quickly, especially during long term experiments. For this reason we suggest a humidifying chamber (Olaf) for incubation.

Directly on an incubated microscope we suggest using Parafilm (Pechiney Plastic Packaging Company) or silicon oil AR 200 (Fluka, Sigma-Aldrich).

Parafilm procedure:
Small stripes of Parafilm on the reservoirs of the slides prevent evaporation effects very well. Fill the slide as recommended. Afterwards, use the Parafilm until it fits tightly.

Silicon oil procedure:
Overlaying the medium with silicon oil prevents all evaporation effects and is cell culture compatible.
Autoclave the silicone oil AR 200 (Fluka, Sigma-Aldrich). Equilibrate oil and cell medium inside the incubator overnight. This step avoids the formation of air bubbles and pre-warms the solutions to 37°C. Afterwards, fill your slide with cells and medium. Overlay the medium’s surface with an appropriate amount of silicon oil. Don’t drip the oil directly on the surface, but let it down run the edges by pressing the pipette tip directly on the upper side of the reservoir. E.g., for µ-Slide VI, fill each reservoir with 30 µl cell-free medium and 30 µl silicone oil.
Please don’t use mineral oil as this would be harmful to the µ-Slides.

_________________________________