Course on Fluorescence Spectroscopy: Principles and Biomedical Applications
In the early 1990s we recognized the expanding use of fluorescence technology in basic biochemistry and biophysics research, and especially the rapid growth in the medical diagnostics and cell imaging. Our discussions with colleagues on campus using our CFS instruments suggested the need for additional training in the many possibilities available from the fluorescence measurements. The same need was found during discussions with several instrument companies. Hence we decided to offer a course in 1993 on the Principles and Applications of Fluorescence Spectroscopy. This course has been offered every year since, until disrupted by Covid. In 2024, we decided to prepare video versions of the lectures which is what you will see below.
These lectures are not intended for a quick glance, but rather to serve as detailed lectures on the principles of fluorescence to working scientists. The course lectures are prepared according to specific topics in fluorescence spectroscopy, and not according to fields of research. The individual lectures are about 30 minutes long and so far there are lectures on:
- Introduction to Fluorescence Spectroscopy
- Instrumentation, Spectra and Quantum Yields
- Introduction to Time-resolved Fluorescence: Time-Domain Measurements
- Introduction to Frequency-domain Time-resolved Fluorescence
- Fluorescence Probes
- Solvent and Environmental Effects on Fluorescence
- Advanced Topics in Fluorescence Resonance Energy Transfer (FRET)
- Quenching of Fluorescence
- Introduction to Fluorescence Anisotropy
- Fluorescence Polarization Assays
In each, we describe the basic principles of the topic, then select biochemical and biomedical examples of the method. The lectures also describe the most common errors in the measurement. The lectures are suitable for a graduate-level course on all aspects of fluorescence.
We are hopeful the lectures may be useful for individual wanting a deeper understanding of fluorescence. We welcome faculty from other institutions to use these lectures as the basis of a graduate course on fluorescence technology. More sections are planned for the future.
Joseph R. Lakowicz, Ph.D.
Baltimore, January 2025
All lectures are available in the Playlist - to locate a specific lecture, select the 'Playlist' menu in the top right corner of the video, then chose the lecture from the sidebar list.
History of the CFS Course
In the early 1990s we recognized the expanding use of fluorescence technology in basic biochemistry and biophysics research, and especially the rapid growth in the medical diagnostics and cell imaging. Our discussions with colleagues on campus using our CFS instruments suggested the need for additional training in the many possibilities available from the fluorescence measurements. The same need was found during discussions with several instrument companies. Hence we decided to offer a course in 1993 on the Principles and Applications of Fluorescence Spectroscopy. The first class in 1993 consisted of about 40 Ph.D. scientists and engineers from instrumentation companies. The scientist’s goal were to design better experiments. The engineer’s goal was to learn the possibilities and to guide instrument development in their companies.
In 1994, we introduced a new component to the course. In addition to lectures, we designed hands-on experiments which could be performed by the course attendees with our own instruments. The hands-on experiments were very popular with the attendees. The range of instruments in the course was expanded by inviting companies to bring in their instruments and also perform demonstration experiments. By 1997, we no longer had the space needed for 70 attendees plus instruments from about 10 companies. All types of fluorescence instruments were brought into the laboratory including steady-state spectrofluorometers, instruments for time-resolved fluorescence in both the time and frequency domains, and microplate readers. We were pleasantly surprised when a couple of microscopy companies brought in a laser-scanning confocal microscopes and a microscope for single molecule detection.
As one can imagine there was considerable disruption of our own research. Because of the combined number of attendees and company personnel the course lectures were moved to nearby hotels and the attendees came to my laboratory for the hands-on experiments. By 2005, we had over 150 attendees and the course. This was before Power Point. Ring binders with several hundred pages needed to be prepared for each attendee.
To assist us with this work, and to bring the CFS course to Europe, one company (PicoQuant in Berlin, Germany) made us an offer we could not refuse. They offered to provide all the facilities and staff support for the CFS course starting in 2006. The only requirement was for me to travel to Berlin to present the lectures. We insisted all instrument companies be welcomed to exhibit their instruments. There were no contracts involved in this process and no funds paid to anyone for the arrangement. It was a pleasant experience to make agreement by a simple handshake and not pages of contracts.
I participated in the course which extended to 2021, when it was interrupted by the COVID epidemic. As a result I gave my lectures by Zoom in 2021 but I found this difficult, without attendee participation. The questions and answers were always the best part of the course.
During the years from 1993 to 2021 the course was always filled to capacity of about 90 attendees int the USA and in Europe limited to 70 attendees. Most of the attendees were at the post-doc level or higher. Attendees at the course included a Nobel Lauret, several members of the National Academy of Science, and a President of the University of Maryland, Baltimore. Over the duration of the course we trained over 2,000 individuals.
In 2024 we decided to prepare video versions of my lectures. Preparation for these video lectures proved to be difficult. In person it was easier to explain concepts even if the slides were not perfect. This is not possible in a video format. There are about 2-fold more slides and more detailed information in these video lectures than in-person lectures. My video lectures include only my slides from the parts of the course, and do not include material from PicoQuant or other lectures.