Using the Core

FLOW CYTOMETRY AT SDSU                                                                                        September 2^nd 2008

 

Dear colleagues,

 

In early October 2005 the Becton Dickinson FACS Aria flow cytometer/sorter was installed in LS316A, and was obtained via the funding of a NSF Major Research Instrumentation grant application with Kathie McGuire as the PI, and David Lipson, Forest Rohwer, Anca Segall, and Constantine Tsoukas as co-PIs.

 

This instrument has been running and available for use since. As the contract with BD is near to its end (September 2007), a renewed management policy will now be installed to ensure the quality and performance of the instrument and, more importantly, the proper training of its users. The instrument cost was ~$330,000. It is an extremely sensitive machine with very expensive parts. Misuse of the machine can damage it, and, from the scientific point of view, distort your results without you even knowing it!

 

The people who have contributed to the writing of the grant have a vested interest in keeping this instrument in top shape for their research and yours, and had formed a committee to oversee its use and maintenance. The following protocol for ALL USERS of the instrument is an updated version based on the recommendations given by the original committee supervising the machine (date of February 7^th 2005).

 

 

Mandatory training before use of the instrument for flow cytometry:

 

1) Basic training in principles of flow cytometry, basic data analysis and the use of flow cytometers must be obtained by ALL current and future users at TSRI. The TSRI Flow Cytometry core offers a monthly 1-day long training session on the first Thursday of every month. To sign up, go to www.scripps.edu, follow the flow cytometry core links and sign up. So far, the TSRI folks have not charged us for this (but this may change depending on how many of our people they train).

We strongly recommend that, before anyone attends the Scripps workshop, potential users should also go through the online training material at

http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/

provided by Becton-Dickinson.

You can always invite the TSRI experts for personal/group training in the instrument’s use and/or data analysis for an additional cost (~$200/h). This will not substitute for the basic training course at TSRI.

2) FACS Aria training: After the basic training requirement has been fulfilled, the future user will have to sign up for a hands-on training session with Brett Hilton, the graduate assistant (GA) in charge of training users on the FACS Aria and with the maintenance of the instrument. Brett Hilton can be contacted by Email (brettjhilton@gmail.com) or phone (619-594-5353). During this session, the trainee can bring her or his own samples to the session and get some practice using these samples. Because such samples are not guaranteed to work, Brett will provide samples he knows are appropriate, and therefore he needs 1 week advance notice to schedule the training session. Exceptions to this should be rare, and at Brett's discretion. A faculty member (Roland Wolkowicz) is in charge of overseeing the instrument and the GA.

3) Data analysis: The longest learning curve is the use of the software, which can be done basically by using the instrument, getting data sets, and reading the manual, playing with the options available, and asking Brett for advice/help. I (Roland Wolkowicz) might be able to consult with you and your students for your analysis needs. Current Protocols in Cytometry are available and are in the hands of Brett. Other resources are the TSRI Flow Cytometry Core web pages and the resources listed there. 

4) Sorting: Brett can help you with the sorting set-up for your experiment. If you need to sort cells, you can always invite the TSRI flow cytometry experts (~$200/h). If you become an experienced user (see below) you will be able to sort by yourself.

All users:

All users, whether old or new, need to introduce themselves to me so I can add them to the list of SDSU ARIA users. This is not meant to intimidate but rather to keep track of users (and familiar faces) for safe use of the machine. I envision three types of users:

 

 The ‘light user’. This user will need to use the machine very few times. This person will not be allowed to use the machine by her/his own. The GA will be present at all times, but remember, the GA is not your personal technician. 

 The ‘dependent heavy user’. This user is not savvy enough to use the machine but needs it often. As with the ‘light user’, you will not be able to use it without the help of the GA. Use common sense, if you feel that the GA is becoming your FACS supporter, give her/him credit for that (for example, authorship or payment for the GA’s time, if extensive). In order to use the machine in the absence of the GA, please see the requirements to become an ‘independent heavy user’.

 The ‘independent user’. This user wants to become independent, whether heavy or light user. To become so, you need to invite (you or up to four of you) Alan Saluk for a personal training in front of the machine (payment will be provided by PIs involved, ~$200/h). Following the personal training, you will perform experiments in the presence of the GA. Only after the GA/myself has recognized your ability to use the machine by your own, you will become an ‘independent user. Again, your name will have to appear on the list of users. It is your responsibility to make sure that this is so.

 

Reminder: all three categories need the basic training at TSRI.

 

The software on the instrument logs the amount of time a user is logged in.  Please be conscious of your time spent logged in. This will be used to track the usage of the machine for payment purposes and in case of (hopefully never-happening) technical emergencies.

 

Signing up for time

 

To sign up for a designated time to use the instrument, go to the SDSU FACS website at www.sdsu.edu/facs/calendar and log in to schedule time.  Please include your name, lab and expected time of use.  This is meant to help you with your needs, especially when a sort/sterile experiment will be performed.   When allotting a time for your experiment, please consider other users and be as accurate as you can with the time you will need on the machine.  Please take in to account your preparation time.

User Fees

The fees for using the instrument are as follows:            

              SDSU Users: $31/hr

              Non SDSU Users: $75/hr

Labs also have the option to pay pre-paid hours as well as other possibilities.  Please see Dr. Wolkowicz or Brett Hilton for details. 

 

This instrument is a major addition to our programs and the quality of research in our University. I foresee an increasing numbers of users that will want to take advantage of it. Please help us keep it in top shape so that we can use it for many fruitful years to come.

 

Roland Wolkowicz, PhD

The FACS Facility Director

 

Anca Segall, PhD

Kathie McGuire, PhD

Constantine Tsoukas, PhD

David Lipson, PhD

Forest Rohwer, PhD

FACS Core Facility at SDSU