The Y2 plate is optimized for time-lapsed imaging of yeast cells with solution exchange. The unique microfluidic cell trapping region holds yeast cells in a uniform focal plane for time-lapse cell microscopy during perfusion flow. Two independent flow units (with identical flow properties) allow simultaneous imaging of two sets of cell/medium combinations. Also see Y16 microfluidic plate.

Key Features:

• Microfabricated cell trap ensures single focal plane
• Switch between 2 medium solutions
• Perfusion culture on your microscope for 3+ days
• Two fully independent perfusion units
• Pipet friendly sample wells
• #1.5 glass bottom for optimal imaging quality

Ordering Info
Instructions
Technical Note

Figure 1. Well Layout

The Y2 microfluidic plate offers the most advanced technology for live imaging of yeast cells. It is the only product that combines the ability to trap yeast in a fixed focal plane with long term continuous perfusion and real time solution exchange.

The easy to use format and fool-proof operation allow any user to run complex live cell imaging experiments with confidence.

The well layout of the Y2 plate is schematically depicted in figure 1. Each flow unit consists of 5 wells arranged in a single row. There are two flow inlets for solution switching, a cell inlet, an open well for imaging, and a flow outlet.

The two units can be operated simultaneously, allowing comparison of two different experiment conditions with identical flow properties.

A #1.5 thickness glass bottom enables high NA imaging on an inverted microscope. The microfluidic plate will fit to any standard 96-well stage holder.

Validated for S. cerevisiae and S. pombe. Please contact info@cellasic.com to check on the suitability of other cell types.

The microfluidic cell culture area is depicted in figure 2. The cells are loaded in a large (1mm x 3mm) rectangular trapping area. This region consists of three 1mm x 1mm trapping pads of progressively smaller ceiling heights for optimized loading of cells (4-6 micron for haploid, 5-8 for diploid). After loading, cells are held firmly in place by the elastomeric ceiling.

Nine (3x3) position markers are incorporated into each trap area to facilitate image navigation. Each marker has a missing dot at the location corresponding to the global position in the array and a "I" or "II" mark to indicate unit number.

A key feature of the Y2 design is the ability to change the solution exposed to the cells in real time. After switching the flow on the control panel, the solution in the cell trapping area will completely turn over as reported in Figure 5. Because of the highly laminar flow profile, a sharp boundary interface between the two solutions will "sweep" across the chamber without forming a mixing gradient. Cells closer to the flow inlets will experience proportionately faster exchange rates.

The expertly designed microfluidic network ensures rapid laminar flow exchange, continuous flows for over 3 days without refilling, and elimination of cross-flows between the exposure channels.

Figure 2. Cell Culture Area

Figure 3. Cell Trapping Mechanism

Figure 4. Flow Rate

Figure 5. Switch Time

Figure 6. S. cerevisiae in the microfluidic device
Courtesy of the Lim Lab, UCSF

Figure 7. S. pombe in the microfluidic device
Courtesy of the Forsburg Lab, US

Figure 8. 3-D Reconstruction of live yeast cells expressing a mitochondrial targeted RFP, imaged on a Spinning Disk Confocal microscope at the UCSF Nikon Imaging Center. Courtesy of Susanne Rafelski of the Marshall lab, UCSF.