UNM Shared Flow Cytometry Resource

HyperCyt®

The HyperCyt® high-throughput flow cytometry platform interfaces a flow cytometer and autosampler. As the sampling probe of the autosampler moves from one well to the next of a multi-well microplate, a peristaltic pump sequentially aspirates sample particle suspensions from each well. Between wells, the continuously running pump draws a bubble of air into the sample line. This results in the generation of a tandem series of bubble-separated samples for delivery to the flow cytometer. Sample and bubble volumes are determined by the time that the autosampler probe is in a microplate well or above a well intaking air. Accurate quantitative measurements have been demonstrated in endpoint assays at rates of 20 to 40 samples/min over a 4-decade range of fluorescence intensity using input cell concentrations of 1-20 million/ml and source well volumes of 5-15 microL. Typical sample volumes of 1-2 microL allow scarce quantities of test cells or reagents to be efficiently used.

As the air bubble-separated samples are delivered in a continuous stream to the flow cytometer, the data are likewise collected in a continuous stream, the accumulated data from all wells of a microplate representing a single data file. The time-resolved data, with periodic gaps corresponding to the passage of the sample-separating air bubbles, are analyzed by proprietary software developed at UNM. The original analysis software, FCSQuery©, has now been superseded by IDLeQuery©, a more advanced analysis package that takes advantage of powerful array processing algorithms, graphics support and multi-platform compatibility features of the IDL programming language (RSI, Inc.).

HyperCyt® Literature

  1. Kuckuck, F. W., Edwards, B. S., and Sklar, L. A. High throughput flow cytometry. Cytometry, 44: 83-90, 2001.
  2. Jackson, W. C., Bennett, T. A., Edwards, B. S., Prossnitz, E., Lopez, G. P., and Sklar, L. A. Performance of in-line microfluidic mixers in laminar flow for high-throughput flow cytometry. Biotechniques, 33: 220-226, 2002.
  3. Jackson, W. C., Kuckuck, F., Edwards, B. S., Mammoli, A., Gallegos, C. M., Lopez, G. P., Buranda, T., and Sklar, L. A. Mixing small volumes for continuous high-throughput flow cytometry: Performance of a mixing Y and peristaltic sample delivery. Cytometry, 47: 183-191, 2002.
  4. Ramirez, S., Aiken, C. T., Andrzejewski, B., Sklar, L. A., and Edwards, B. S. High-throughput flow cytometry: Validation in microvolume bioassays. Cytometry, 53A: 55-65, 2003.
  5. Waller, A., Simons, P., Prossnitz, E. R., Edwards, B. S., and Sklar, L. A.High throughput screening of G-protein coupled receptors via flow cytometry. Comb Chem High Throughput Screen, 6: 389-397, 2003.
  6. Edwards, B. S., Oprea, T. I., Prossnitz, E. R., and Sklar, L. A. Flow cytometry for high throughput, high content screening. Curr Opin Chem Biol, 8: 92-396, 2004.
  7. Young, S. M., Curry, M. S., Ransom, J. T., Ballesteros, J. A., Prossnitz, E. R., Sklar, L. A., and Edwards, B. S. High-throughput microfluidic mixing and multiparametric cell sorting for bioactive compound screening. J Biomol Screen, 9: 103-111, 2004.
  8. Bartsch, J. W., Tran, H. D., Waller, A., Mammoli, A. A., Buranda, T., Sklar, L. A., and Edwards, B. S. An investigation of liquid carryover and sample residual for a high-throughput flow cytometer sample delivery system. Anal Chem, 76: 3810-3817, 2004.
  9. Waller, A., Simons, P. C., Biggs, S. M., Edwards, B. S., Prossnitz, E. R., and Sklar, L. A. Techniques: GPCR assembly, pharmacology and screening by flow cytometry. Trends Pharmacol Sci, 25: 663-669, 2004.
  10. Young, S. M., Bologa, C., Prossnitz, E., Oprea, T. I., Sklar, L. A., and Edwards, B. S. High throughput screening with HyperCyt flow cytometry to detect small molecule formylpeptide receptor ligands. J Biomol Screen, in press, 2005.