Courses Overview

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 Salem Haile.

Basic Flow Cytometry Course

Who should take this 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.

What benefits does it provide?

• 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.

Theoretical Tutorial Outline

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

Hands-On Tutorial Outline

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

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.

Registration and Times

To register for this course please contact Salem Haile. 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. 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. A minimum of four confirmed students is needed to arrange a suitable class date. The maximum class size is eight students.

Nucleic Acid Flow Cytometry Course

Who should take this course?

• 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.

Theoretical Tutorial Outline

1. Brief Introduction to Flow Cytometry
2. Fundamentals of Nucleic Acid Analysis
   • Cell Cycle Phases
   • The DNA Histogram
3. Stem Cell "side population"
4. Cell Cycle Kinetics
   • Pulse processing
   • Nucleic Acid Stains and their Properties
5. DNA/RNA Measurements
6. Proliferation
   • Bromodeoxyuridine
   • EDU Click-iT Technology
   • Pulse-Chase Experiment, Cell Cycle Arrest
   • Dye Dilution Assay
7. Ploidy
8. Chromosome Analysis
9. Measurement of Apoptosis and Dead Cell Discrimination
10. Combination of Nucleic Acid with Cell Surface Antigen Analysis
11. Combination of Nucleic Acid with Intracellular Protein Analysis

Hands-On Tutorial Outline

1. Sample Acquisition for DNA content analysis (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

Course Fees

Compared to commercial courses, the UCLA Nucleic Acid and Proliferation 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.

Registration and Times

To register for the nucleic acid course please call Salem Haile. 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.