The flow cytometry and cell sorting facility provides ISB scientists and affiliated companies with state-of-the-art flow cytometric analyses and high-speed sorting of rare cells from complex samples.
Services offered:
- Three-color flow cytometry using a BD FACSCalibur
- High-speed cell sorting (up to 24,000/sec) logically gated on up to 16 parameters
- Magnetic cell sorting using a Miltenyi autoMACS
- Choice of up to 8 different excitation wavelengths
- Consultation for novel cytometric assays
- Complete fluorescence emission reconstruction for novel fluorochromes
- Training for use of the BD FACSCalibur
Equipment and procedures:
The facility has a Cytopeia inFlux high-speed cell sorter as well as the prototype instrument for the commercial version, a BD FACSCalibur three-color flow cytometer, and a Miltenyi autoMACS magnetic cell separator. The cell sorting instruments can be custom configured to use a range of fluorophores and excitation wavelengths.
Example(s) of projects that use the services of the facility:
- Deciphering the regulatory network of the archaeon, Halobacteria NRC-1
- Cytometric measurements of the GAL regulon to quantify transcriptional variation in a population
- Linking phylogeny to function to determine complex community structure in extreme environmentsFlow cytometric determination of the identity of the prostate stem cell. Cytometric identification and monitoring of toxic marine diatoms responsible for harmful algal blooms (HABs)Cytometric determinations of sexual reproduction in marine diatoms.
Representative publication(s):
Gold, E.S., R.M. Simmons, T.W. Petersen, L.A. Campbell, C.C. Kuo, and A. Aderem 2004. Amphiphysin Iim is required for survival of Chlamydia pneumoniae in macrophages. J.Exp.Med. 200: 581-586.
Lansdorp, B.M., P.I. Nelson, T.W. Petersen, and G. van den Engh 2004. An optical monitor for measuring the amplitude and phase of oscillations on the surface of a capillary jet in a high speed cell sorter. Review of Scientific Instruments 75: 741-748.
Matsumoto, M., D. Einhaus, E.S. Gold, and A. Aderem 2004. Simvastatin augments lipopolysaccharide-induced proinflammatory responses in macrophages by differential regulation of the c-Fos and c-Jun transcription factors. J.Immunol. 172: 7377-7384.
Orellana, M.V., T.W. Petersen, and G. van den Engh 2004. UV-excited blue autoflourescence of Pseudo-nitzschia multiseries (Bacillariophyceae). J.Phycol. 40: 705-710.
Petersen, T.W., S.F. Ibrahim, A.H. Diercks, and E.G. van den 2004. Chromatic shifts in the fluorescence emitted by murine thymocytes stained with Hoechst 33342. Cytometry 60A: 173-181.
Petersen, T.W. and G. van den Engh 2003. Stability of the breakoff point in a high-speed cell sorter. Cytometry 56A: 63-70.
Ramsey, S., D. Orrell, J.J. Smith, M. Marelli, T.W. Petersen, P. de Atauri, J.D. Aitchison, and H. Bolouri 2005. Dual feedback loops in the GAL regulon suppress cellular heterogeneity in yeast. In preparation.
Uy, J.L., C.L. Asbury, T.W. Petersen, and G. van den Engh 2004. The polarization of fluorescence of DNA stains depends on the incorporation density of the dye molecules. Cytometry 61A: 18-25.
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