Bloodworks Research Institute
Flow Cytometry Lab
The Flow Cytometry Laboratory (Flow Lab) provides cell sorting and analysis services for in-house and external investigators. The Flow Lab staff operates the sorter and assists investigators using the analyzers. The staff also provides consultation regarding cytometry experimental design, sample preparation and data analysis. In addition, the staff welcomes the opportunity for external research project contracts.
The Flow Lab is equipped with FACSAria cell sorter, LSR II cytometer and Amnis ImageStream Mk II imaging cytometry system. The lab is directed by cell analysis specialist Dr. Xiaoping Wu. Prior to joining BloodworksNW Research Institute, Dr. Wu had extensive research training in Immunology and Molecular Biology.
The Flow Lab has been actively involved in ongoing and previous projects addressing bleeding disorder, transfusion complications, coagulopathy in traumatic brain injury, sickle cell disease, and thrombosis, megakaryocytopoiesis, platelet generation, autoimmunity and therapeutic immune modulation for inhibitory antibodies against factor VIII.
The Lab is located in Room 1136 in the Research Institute of the BloodworksNW at 1551 Eastlake Ave E, Seattle, WA 98102.
Charges
- $130/hour for cell sorting by FACSAria
- $90/hour for analysis on LSR II
- $250/hour for analysis on Amnis ImageStream Mk II
Biosafety
The staff requires information about the use of infectious agents, sample sources and preparation procedures before accepting samples for processing in the Flow Lab.
Reservation
Service can be reserved via phone (206) 568-2238, lab (206) 568-2162 or email [email protected].
Reserved service can be canceled no later than 3 hours in advance of the scheduled time. If not canceled, half of the fee for the reserved time will be charged.
Cytometer Specifications
| Cytometers | Lasers | Commonly Detected Dyes |
| FACSAria cell sorter | 488nm, 633nm, 561nm and 405nm | FITC, PE, PerCP-Cy5.5, APC, Alexa 700, APC-Cy7, PE-TexRed/mcherry, PE-Cy5, PE-Cy5.5, PE-Cy7, Pacific Blue, AmCyn, Qdot605 and Qdot655 |
| LSR II analyzer | 488nm, 633nm, 405nm and 355nm | FITC, PE, PerCP/PE-Cy5/PerCP-Cy5.5, PE-Cy7, APC, Alexa Fluor700, APC-Cy7, Pacific Blue, AmCyn, BV605, Indo-Violet and Indo-Blue. |
| Amnis ImageStream Mk II | 488nm, 561nm, 642nm and 405nm | FITC, PE, PE-TexRed/m-cherry, PerCP-Cy5.5, PE-Cy7, Pacific Blue/DAPI, AmCyn, efluor625/BV605, APC and APC-Cy7 |
Representative Publications
- Su RJ, Epp A, Feng J, Roy J, Latchman Y, Wu X, Bolgiano D, Josephson NC. Suppression of the immune response to FVIII in hemophilia A mice by transgene modified tolerogenic dendritic cells. Mol Ther. 1896-904, 2011. PMID21772255
- Wang W, Gilligan DM, Sun S, Wu X, Reems JA. Distinct functional effects for dynamin 3 during megakaryocytopoiesis. Stell Cells Dev. 2139-51, 2011. PMID21671749
- Lin Y, Wu X, Shen Y, Bu P, Yang D, Yan X. A novel antibody AA98 V(H)/L directed against CD146 efficiently inhibits angiogenesis. Anticancer Research. 27: 4219-24, 2007 PMID18225593
- Komori A, Xu Z, Wu X, Zan H, Casali P. Biased dA/dT somatic hypermutation as regulated by the heavy chain intronic iE enhancer and 3’E enhancers in human lymphoblastoid B cells. Molecular Immunology, 43: 1817-26, 2006 PMID16412510
- Wu X, Tsai CY, Patam MB, Zan H, Chen JP, Lipkin SM, Casali P. A role for the mismatch repair Mlh3 protein in immunoglobulin class switch DNA recombination and somatic hypermutation. The Journal of Immunology, 176: 5426-37, 2005 PMID16622010
- Kim EC, Edmonston CR, Wu X, Schaffer A, Casali P. The HoxC4 homeodomain protein mediates activation of the immunoglobulin heavy chain 3′ hs1,2 enhancer in human B cell: relevance to class switch DNA recombination. Journal of Biological Chemistry 279: 42258-69, 2004. PMID15252056
- Wu X*, Zan H*, Komori, A, Holloman W, Casali P. CD40-induced and AID-dependent generation of resected double-strand DNA breaks and recruitment of Rad52/Rad51 in somatic hypermutation. Immunity 18: 727-738, 2003. (*Equal contributor) PMID12818155
- Wu X, Feng J, Komori A, Kim EC, Zan H, Casali P. Double strand DNA breaks, AID and error-prone DNA synthesis in Immunogobulin somatic hypermutation. Journal of Clinical Immunology 23: 235-246, 2003 PMID12959216
- Schaffer A, Kim EC, Wu X, Zan H, Testoni L, Salamon S, Cerutti A, Casali P. Selective inhibition of class switching to IgG and IgE by recruitment of the HoxC4 and Oct-1 homeodomain proteins and Ku70/Ku86 to newly identified ATTT cis-elements. Journal of Biological Chemistry 278: 23141-23150, 2003 PMID12672812
Lab Photos
Dr. Xiaoping Wu, cell analysis specialist of the Flow Cytometry Core, is conducting experiments on our flow cytometers.
Funded by M. J. Murdock Charitable Trust, Dorothy F. Thorne Foundation, Seattle Foundation Medical Funds and BloodworksNW Faces For Life events, our high-performance FACSAria cell sorter, the LSR II analyzer and two other cytometers, FACSCalibur and FACSVantage, are essential tools for BloodworksNW researchers in thrombotic and bleeding disorders, and transfusion medicine.
Amnis ImageStream Mk II
BD FACSAria cell sorter
BD LSR II cytometer
Xiaoping Wu, PhD
Cell Analysis Specialist
Lab Manager
Bloodworks Northwest
Research Institute
1551 Eastlake Avenue E, Suite 100
Seattle, WA 98102
Email: [email protected]
Phone: (206) 568-2238
Lab: (206) 568-2162
FAX: (206) 587-6056