Flow Cytometry
 

Facilities:

The VGTI Florida® Flow Cytometry core facility provides users with state of the art instrumentation and services. The principal assets of the core consist of three analytic instruments (two BD LSR II instruments and one BD FacsArray) and two sorting instruments (BD Aria II). The core also provides data analysis support and operates four Apple workstations with FacsDiva and FlowJo software. 

Sorting Facility

The sorting facilities of the core are housed in two rooms, one dedicated for the sorting of uninfected samples (room 314C) and another for the sorting of infected samples (room 135K). The BD Facs Aria II is a flexible and highly adaptable bench top sorter.  It contains four fixed-aligned, air-cooled lasers: red (635nm), green (532nm), blue (488nm) and violet (405nm).  In its present configuration, the sorter is capable of performing 18 parameters (forward and side scatter plus 16 color flow cytometry). The Aria II has an increased sensitivity and flexibility due to the octagon and trigon optical arrays.  
 
It can yield more information from each sample and rare event analysis is more accurate and efficient because of the increased flow rate and the digital acquisition system. 

These instruments are equipped with temperature-controlled chambers and have the ability to sort single cells into 96 well plates. Both instruments also have the aerosol management system in place to eliminate the emission of aerosols generated during operation. Further, the Aria II designated for sorting of infected samples will be operated by individuals wearing respirators to further decrease the potential for contact with infectious aerosols. These instruments are only operated by qualified individuals on a pre-scheduled basis.  


 

Analytical Facility

The primary analytic facilities of the core are housed in room 318B, which contains 2 BD LSR II analytic instruments. The BD LSR II is a flexible and highly adaptable bench top analyzer. It contains four fixed-aligned, air-cooled lasers: red (635nm), green (532nm), blue (488nm) and violet (405nm). In its present configuration, the analyzer is capable of performing eighteen parameters (forward and side scatter plus sixteen color flow cytometry). The LSR II has an increased sensitivity and flexibility due to the octagon and trigon optical arrays. It can yield more information from each sample and rare event analysis is more accurate and efficient because of the increased flow rate and the digital acquisition system.
These instruments are equipped with HTS units that allow acquisition of samples automatically from 96 and 384 well plates. Additionally, they are also equipped with BD FACS flow supply systems to automate fluid handling. The core provides training to individuals of the Institute so they are able to operate these instruments unassisted by core staff.
 
The room 318B also contains a two laser, 6 parameter BD FacsArray. This instrument is dedicated to performing multiplex bead assays and high throughput screening experiments. The core will provide training to individuals so they can operate this instrument without core assistance.
 
Services:
The flow cytometry core will provide the following services:
  1. Sorting of infected and uninfected samples.
  2. Analysis and acquisition of flow cytometry data.
  3. Training on the use of flow cytometry technology and equipment.
  4. Assistance in the development of flow cytometry staining panels.

Advice and consent on experimental design.


Personnel:
John Schatzle Ph.D. (Director of Scientific Operations)
(772) 345-5718
 
Yu Shi Ph.D., MD (Research Associate, Core Facility Coordinator and Technical Specialist)
(772) 345-5722
 
Kim Kusser (Senior Research Assistant)
 
Zong He Ph.D. (Research Associate)
 
Sample Submission:
These services will be scheduled by contacting Kim Kusser. Fees will be assessed on an hourly rate based on actual usage (for the user initiated analysis and for analysis by the core). Cell sorting will be scheduled in ½ day blocks.

 

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