The UCLA Core facility offers two flow cytometry courses.  One class teaches basic flow cytometry principles in addition to offering hands-on training on the operation of Becton Dickinson analytic flow cytometers.  The other course teaches principles of nucleic acid analysis by flow cytometry as well as hands-on instrument set-up and proper use of the specialized DNA analysis software. In addition, the Core facility provides individual training in the use of the RoboSep magnetic cell sorter which is mandatory for all  new users of the RoboSep. To arrange for this training, please contact Beverly Redsar or Ingrid Schmid. 

WHO SHOULD TAKE THE BASIC FLOW CYTOMETRY COURSE?

• Users of the core facility who plan to acquire their own samples and wish to operate the Becton Dickinson analytic flow cytometers under the supervision of the Core staff. Only graduates of this class are permitted to run their own samples on the instruments.
• Investigators who do not intend to operate a flow cytometer but plan to use flow cytometry in their research. The theoretical tutorial will provide investigators with an overview of flow cytometric basics and will allow them to gain insight into the research potential of flow cytometry techniques.
• All researchers and research associates (within UCLA and off-campus) with access to their own flow cytometers who wish to utilize this training resource.

BENEFITS

• Graduates of this class can acquire samples at a discounted rate compared to the technician-assisted rate on the analytic flow cytometers available in the core facility.
• Graduates not only learn valuable flow cytometry principles and techniques, but they can reserve instrument time regardless of other technician-assisted appointments and continue to utilize the core facility’s resources and personnel expertise.
• Investigators will enhance their insight into flow cytometry as a research tool.

COURSE FEES

Compared to the cytometer manufacturer’s courses, the UCLA Core flow cytometry course is an excellent value for UCLA faculty, staff, and students.  Please, refer to the "services" section for the class fee associated with your membership status. This fee includes pre-course reading material and a course booklet.  If you have to cancel your appointment, please call 24 hours in advance.  If you fail to do so, we reserve the right to charge the entire course fee.

GRADUATE USER FEES

Graduates who run their samples on the instruments available in the core facility receive a reduced rate compared to users who need staff to run their samples (see rate table in "services" section). Core personnel assist course graduates during any appointments subsequent to training (if necessary) at the discounted fee per hour until they are comfortable acquiring samples on the flow cytometer and analyzing data on their own.

COURSE OVERVIEW

These one-day classes, held in the Core facility, begin at 9:00 a.m. and end at approximately 4:45 p.m. There is a one hour lunch break and two short breaks; one in the morning and one in the afternoon.

The two major components of the course are:

1. Tutorial on the technical aspects of flow cytometry, and
2. Hands-on training in the operation of the Becton Dickinson analytic flow cytometers.

The theoretical tutorial gives a brief overview of basic flow cytometric principles including:

1. Components of flow cytometers (fluidics, lasers, filters, detectors)
2. Fluorescence absorption and emission
3. Spectral overlap
4. Fluorescence compensation
5. Fluorescence intensity measurements
6. Data analysis
7. Biosafety recommendations

The hands-on training includes:

1. Introduction to instrument set-up
2. Introduction to CellQuest software
3. Introduction to gating principles
4. Set up of multi-color flow cytometry
5. Electronic compensation adjustments
6. Data analysis techniques
7. Troubleshooting tips
8. A brief overview of Core Laboratory Policies
9. Practical Biosafety considerations

REGISTRATION

To register for this course please call Beverly Redsar at (310) 825-1043 or click name to send an e-mail.  You will be contacted with information about course availability and date.

The course is held on an as-needed basis, usually once every month, most often on Mondays and Tuesdays. A minimum of four confirmed students is needed to arrange a suitable class date.  The maximum class size is eight students.

• Investigators who plan to use flow cytometry to measure DNA content, DNA ploidy, cell cycle kinetics, RNA content, and apoptosis in their research.  The theoretical tutorial provides investigators with an in-depth overview of the basics of this sub-discipline of flow cytometry and allows them to gain insight into the research potential of these techniques.  Prior attendance of the basic flow cytometry course if preferred, but not required.  Please note that the attendance of the nucleic acid course per se does not entitle graduates to independent data acquisition privileges on the analytic flow cytometers.

• All researchers and research associates (within UCLA and off-campus) who would like to learn more about nucleic acid analysis by utilizing this training resource.

Graduates not only learn valuable flow cytometry principles and techniques, but they can utilize the core facility's considerable resources and personnel expertise to enhance their insight into flow cytometry as a research tool.

Compared to instrument manufacturers' DNA analysis courses, the UCLA Nucleic Acid flow cytometry course is an excellent value for UCLA faculty, staff and students. Please, refer to the "services" section for the DNA class fee associated with your membership status.   Fees include pre-course reading material and a course booklet.  If you have to cancel your appointment, please call 24 hours in advance.  If you fail to do so, we reserve the right to charge the entire course fee.

THEORETICAL PART

1. Brief Introduction to Flow Cytometry
2. Nucleic Acid Stains (DNA Specific Stains, Nucleic Acid-Specific Stains, Non-Specific Polyanion (DNA/RNA) Stains)
3. Fundamentals of Nucleic Acid Analysis
   • Cell Cycle Phases
   • The DNA Histogram
   • Cell Cycle Kinetics
   • Bromodeoxyuridine
   • Quenching of Hoechst Fluorescence by BrdU Incorporation
   • Ploidy
   • Cell Death and Apoptosis
   • Chromosome Analysis
   • DNA/RNA Measurement
4. Combination of Nucleic Acid with Cell Surface Antigen Analysis
• Cell Surface Antigens and DNA Content
• Cell Surface Antigens and DNA/RNA Content
5. Flow Cytometer Setup
6. Cell Cycle Analysis Theory (Histogram Deconvolution)
7. Sample Preparation Protocol

PRACTICAL PART

1. Sample Acquisition (Optimizing Settings, Recognizing Aggregates/Debris)
2. ModFit™ Analysis (Used for DNA Histogram Analysis)
3. Analysis of Nucleic Acid Stained Samples with CellQuest™
   • Dead Cell Discrimination
   • Apoptosis
   • Combined Cell Cycle Analysis and Immunofluorescence

To register for the nucleic acid course please call Beverly Redsar at (310) 825-1043 or click on the name to send an email.  You will be contacted with information about course availability and date.

The course is held on an as-needed basis.  A minimum of four confirmed students is needed to arrange a suitable class date.  The maximum class size is eight students.