Frequently Asked Questions

Technical






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How do I detect migrating cells?

You can detect cells that have migrated through the Barrier by adjusting the focus of your microscope onto the bottom of the membrane and/or onto the coverglass. A detailed procedure for conducting and analyzing a migration experiment is available by request

How often should I change the media?

For CytoVu®, the limiting factor for changing the media is evaporation rather than depletion of nutrients. This is due to the small volume of media that is used in the device. To combat evaporation and prolong the amount of time between replacements, we have installed a lid on CytoVu®.

Even with these measures, we recommend that researchers replace the media once daily.

How do I focus on imaging a specific side of the membrane?

CytoVu® gives you the possibility to culture on both sides of the membrane and on the coverslip. To focus on a different surface, merely adjust the z-plane of focus on your microscope.

How do I co-culture with this device?

Briefly, you can co-culture in CytoVu® by pipetting cells into the apical and basal wells. Cells pipetted into the apical well will settle and attach to the apical side of the membrane. Cells pipetted into the basal will settle and attach to the basal side of the membrane after inverting the slide. The cells will then be in perfect communication across the membrane.

Please watch our tutorial on YouTube for more information.

Can I add Matrigel to the membrane?

Matrigel can be added to CytoVu®. Customers typically deposit 4 ul of Matrigel into each well. The small volume requires that you keep the following points in mind while working with Matrigel in CytoVu®:

Keep the Matrigel and all materials/equipment chilled while applying it to CytoVu®.
Work quickly to keep the Matrigel from solidifying early or irregularly.

Why should I pre-coat the wells?

You pre-coat the membranes by pre-incubating media or your ECM component of choice to the membranes before you add your cells. This helps to ensure that the necessary compounds are laid down on the membrane that will allow your cells to attach efficiently.

Can I remove adhered cells from CytoVu®?

Cells that adhered to the membrane can be removed by treating with trypsin or cold PBS. We do not recommend mechanical modes of removing adhered cells because the membrane in CytoVu® is fragile.

Do liquids added to one well transfer between wells?

The media from the apical and basal wells will intermix when there is a monolayer that is not fully congruent. The presence of a complete monolayer will limit diffusion between the compartments to only those molecules that are actively transported across the monolayer or to those that are able to passively diffuse through the monolayer.

How can I get my cells to adhere better?

If your cells are not adhering to the CytoVu® membrane it is most likely that your cells require a layer of ECM proteins in order to attach to the membrane. The CytoVu® membrane is very similar to glass and frequently requires presence of ECM components for cells to attach. Try one of the following to promote adherence.

1. Incubate media in CytoVu® overnight before seeding your cells.
2. Conduct a literature search to determine the ECM components that are required for your cell type to bind to glass. Pre-coat the membrane with these components by incubating overnight.

A second reason why your cells may not be adhering is because too much media evaporates in between replacements. In order to ameliorate this, try to replace the media more frequently or store the CytoVu® slide in a sealed petri dish that also contains wet culture paper.

How can I limit evaporation in CytoVu®?

To combat evaporation, try the following:

1. Make sure the lid is firmly attached.
2. Place the CytoVu® slide in a sealed petri dish before putting it in the incubator.
3. Place a damp piece of culture paper in the petri dish along with CytoVu® slide.

How do I sterilize?

CytoVu® should be sterilized before use. It is recommended that it be sterilized by autoclaving. Remove CytoVu® from all packaging materials and place it in an autoclave-safe container. Autoclave using the standard dry cycle.

Additionally, CytoVu® can be treated with ethanol and/or UV radiation without any negative effects.

How do I make the degradable NanoBarrier™ degrade?

The degradable NanoBarrier™ will begin to degrade following normal exposure to media.