Frequently Asked Questions:
1. What applications are the ONIX™ Platform suited for?
The platform is designed for dynamic flow control during time-lapse imaging of live yeast and mammalian/adherent cells. Typically, any experiment that can be performed with an inverted fluorescence microscope will work on the microfluidic plates. Some example experiments performed by our customers include:
- Gene expression via GFP/YFP/CFP/RFP intensity
- Protein translocation
- Cell cycle analysis
- Intracellular binding via FRET
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2. Is there background fluorescence from the microfluidic plates?
In tests with 60X and 100X oil immersion objective lenses, no significant background fluorescence has been found in the typical wavelengths used for imaging (>300 nm). The materials in contact with the cells are: 1) optical quality glass slide (#1 thickness), and 2) PDMS polymer in the walls of the microfluidics. The optical properties of PDMS are reported to be similar to BoroFloat glass for biological analyses (Piruska, et al, Lab Chip, 2005, 5, 1348-1354).
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3. What yeast cell strains can I use?
The Yeast microfluidic plates are designed to accommodate yeast cells from 4-7 micron in diameter. This has been validated with S. cerevisiae (diploid and haploid) and S. pombe.
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4. Does the cell trapping mechanism affect cell behavior?
The trapping pad is a soft elastomer, analogous to an agar pad, and does not seem to affect cell growth and normal functions.
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5. How long can I run the microfluidic plate in a single experiment?
The small volume of the perfusion circuit means that very low flow rates (on the order of nl/min) are sufficient to rapidly replenish the medium around the cells. The suggested experiment limit is 48 hours, however, longer culture periods are possible.
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6. Can I use the microfluidic platform to observe yeast cell division?
Yes. A key motivation for the design of the cell trap region was to allow improved quantitative analysis of cell division. A major advantage of our method is that even while the yeast cells bud, the daughter cells are kept in the same microscope focal plane. This is not possible with alternative methods such as agar pad and perfusion chambers, where oftentimes the budding cell will move in and out of the focal plane, obscuring quantitative analysis.
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7. What is the minimum sample volume I can use?
Theoretically, the microfluidic plates can be operated with about 1 ul of liquid. However, in order to comfortably cover the bottom of the 96 well format reservoir, we recommend using at least 40 ul of sample per well. Smaller volumes can be used if care is taken.
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8. Can I run the different units of the M8/Y8 plate on different days?
Yes-- however, care must be taken to prevent evaporation of the wells in between experiments. Once the well dries out, it can no longer be used. Therefore, we recommend using multiple Y2/M2 plates instead.
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9. Are the two cell types uniformly exposed in the M2/Y2 plate?
Yes. The design of the microfluidic channels ensures that both units receive identical flow profiles, giving identical exposure to the two populations.
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10. How uniform are the flows if I run all 8 units of the M8/Y8 plate in one experiment?
Flow lag (if any) between the eight units of the plate is less than a few seconds. This is due to the careful design and fabrication of the microfluidic channels, as well as the nature of laminar flows.
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11. Where can I get additional fittings to adapt my air/vacuum inlets to the control panel?
If your supply line does not fit to the included 3/8" barbed adaptor, please find the appropriate fittings via your preferred hardware vendor. CellASIC recommends McMaster-Carr (http://www.mcmaster.com).
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12. Do I need vacuum and/or compressed air line nearby to operate the system?
The ONIX micro requires lab air and vacuum to operate.
The ONIX Standard and Advanced versions contain internal pumps for pressure and vacuum.
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13. Is there a warranty on your products?
If for any reason you find a defect in a product or are unsatisfied with a product ordered from CellASIC 6 months from the date of purchase, we will provide a replacement for you as soon as possible.
Contact Customer Service at:
customerservice@cellasic.com
Toll Free: 1-866-316-1259
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