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Department of Tropical Medicine & Hawaii Center for AIDS – Flow Cytometry Core Facility
Overview:
The Department of Tropical Medicine and Hawaiʻi Center for AIDS offers valuable resources in the area of analytic flow cytometry. The mission of the Flow Cytometry core is to provide these services to scientists and researchers affiliated with the University of Hawaiʻi’s (UH) multidisciplinary research program and non-UH users on first come first served fee for service basis. Users can be trained to use the flow cytometer equipment by experts within the Core.

Location:
Room 324
Biosciences Building
John A Burns School of Medicine
651 Ilalo St, Honolulu, HI 96813
(Parking available)

Contacts:

Flow Core Manager: Christopher K. Sugai, MSc csugai@hawaii.edu
Administrator: Jason Huynh huynhjas@hawaii.edu
Director: Lishomwa Ndhlovu, MD, PhD lndhlovu@hawaii.edu
Phone: 808-692-1604

Services and Instrumentation:
The core maintains a BD LSR Fortessa Flow cytometer instrument, computer work-stations for data analysis, and related equipment to support research requiring molecular marker characterization of cells in suspension.

BSL2 safety practices should be employed in the facility.

Availability: 24 hours, 7 days a week.

The BD Fortessa: This instrument is operated using FACSDiva Software and are available for all labs to use. This flow cytometer analyzer is equipped with 3 or more lasers. Analysis occurs at the cuvette flow cell, increasing sensitivity and resolution, allowing detection of 2 relative physical properties as well as up to 17 different colors simultaneously. It can be run at 3 different sample flow rates with a fine-tune adjustor and is capable of analysis rates of up to 40,000 events per second.

All BD Fortessa users are required to take Basic Training

Applications:

• Some flow cytometric applications that can be analyzed in the flow cytometry facility include:
• Immunophenotyping
• Cell Cycle
• Ploidy
• Mitochondrial Potential
• Apoptosis
• PhosFlow
• Live/Dead
• Stem Cell Analysis
• Cell Proliferation
• Oxidative Burs

Training:
You must receive Basic Training before utilizing any of the self-use equipment in the Flow Cytometry Core. This will cover Core logistics, flow cytometry basics, basic experimental design, instrument startup/utilization, workflow, FACSDiva software overview, running test samples, system changeover/shutdown, analysis options, data management, and general troubleshooting. The Core will provide all necessary reagents and samples for the training.

New Users: schedule a minimum of 3 hours.
Refresher: schedule a minimum of 1.5 hours.

To schedule Basic or Advanced Training please contact the Flow Core Manager. Groups of three can be accommodated per session upon conformation on the availability of a Core staff member to train you.

Sample Preparation:
The recommended sample volume is no less than 0.3 ml. Samples should be resuspended in 12 x 75 mm FALCON 5 ml tubes (polystyrene Falcon# 352052). If you think your tube may not fit properly, please ask for the tubes to transfer your sample. Samples should be fixed in 0.5-2% paraformaldehyde or methanol- free formaldehyde to kill infectious material. Do not use common laboratory grade formaldehyde. Infectious material should be fixed in paraformaldehyde for at least 30 minutes for best results. Please, no unfixed infectious material or radioactive material. Human samples must be fixed or have been tested to show that they are noninfectious.

Filter cells that are sticky or clumping with nylon mesh (Falcon 5 ml Polystyrene Round Bottom Tube with Cell-Strainer Cap 12 x 75 mm; Part number 352235). Anything less than 70 microns should pass easily through the cytometer. However, this assumes a single cell suspension. Cells that grow in monolayers, as well as large populations of dead cells or debris, will cause samples to clump. Consider adding heparin to the cell suspension if the cells are extremely sticky.

Rates and Billing
Recharge rates are available upon request. Please inquire with the Flow Core Manager.

*Disclaimer: Upon our evaluation we reserve the right to regulate access to the instruments based on the users ability and proper use of the instrumentation in the Flow Core.