The 3D:M plate enables long term perfusion culture of cells in a three dimensional gel. Load your cells and gel into the micro-chamber via a passive capillary loading method. Cells are fed by gravity perfusion, thus requiring no additional equipment (besides a pipette) to operate this advanced microfluidic plate.

Key Features:

• Twenty four fully independent perfusion units
• Gel confined to a 150 nL micro-chamber
• Perfusion enables long term 3D culture
• Pump-free culture in a standard incubator
• Pipet friendly sample wells
• #1.5 glass bottom for optimal imaging quality

Ordering Info
Instructions
Technical Note

Figure 1. Well Layout

The 3D:M microfluidic plate offers the most advanced technology for 3D cell culture experiments. Our pioneering microfluidic design allows long term culture of cells in a gel while providing continuous perfusion exchange.

The easy to use format and fool-proof operation allow any user to run long-term 3D cell culture experiments with confidence.

The well layout of the 3D:M plate is schematically depicted in figure 1. Twenty-four flow units are tiled on a 96 well plate frame. Each flow unit consists of 4 wells arranged in a single row, representing the flow inlet, the cell culture chamber, the cell/gel inlet, and the flow outlet.

The fluidics are operated using passive forces, such as gravity and surface tension, eliminating the need for any external hardware. All that is necessary to run the 3D:M plate is a pipette and an incubator.

A #1.5 thickness glass bottom enables high NA imaging on an inverted microscope. The defined 200 micron thick 3D gel layer significantly improves imaging quality compared with traditional gel culture methods.

A set of air channels allows gas diffusion into the cell culture area and upstream flow channels (light blue). Due to the high gas permeability of the materials and rapid microscale diffusion, this innovative design maintains a well balanced oxygen and CO₂ supply to cultured cells.

The microfluidic cell imaging design is depicted in figure 2. The culture chamber is 1.5x0.5x0.2 mm in size. Cells are introduced via a passive capillary flow method as depicted in figure 3. As cells travel down the streamlines (black arrows), they are loaded into the culture chamber, assuring a low pressure, low stress culture environment.

Cells can be loaded mixed with a gel of your choice (embedded) or with the gel loaded after cell introduction (overlay). The chambers can also be used in the absence of gel for a 2D culture.

After the cells/gel are loaded, they are fed by perfusion of media from the inlet well. The perfusion barrier surrounding the gel chamber has 2 micron pores to allow free diffusion of liquids into and out of the gel chamber (figure 4). The barrier also prevents cells/gel from blocking the flow channel.

The continuous perfusion enables cultured cells to be studied for long term experiments (figure 5). The short diffusion distance of the micro-chamber provides turnover on the order of 20 minutes. The gravity perfusion rate is set at 40 microliters per day, allowing continuous flow for up to 3 days before refilling the inlet is necessary.

The small volume of the microfluidic chamber makes this method especially useful for limited cell/gel samples and primary cells.

Suitable for adherent and non-adherent cell types. Please contact info@cellasic.com if you have questions regarding a particular cell type.

Figure 2. Cell Culture Area

Figure 3. Capillary Cell Loading Mechanism

Figure 4. Gravity Perfusion

Figure 5. 3D Morphology of Cells in Gel