This week I attended the 1st South East Asian Course on “F-techniques: FCS, FCCS, FLCS, FRET, FLIM, and FRAP.” The course was dedicated to a variety of fluorescent microscopy techniques and their applications in the life sciences. The organizers for the workshop were the Institute of Medical Biology Microscopy Unit, EINST Technology Pte Ltd. (both from Singapore) and PicoQuant GmbH (Germany).
The first day of the course was dedicated to an in depth introduction to the techniques. I began the course with a ground level understanding of fluorescent microscopy so I felt completely exhausted after the first day of introduction! Then again, the fact that I was listening to presentations from 9:00 to 6:00 may have also had something to do with that (the course provided ample amounts of coffee). Many of the following days were applications of these techniques in scientific research (all days were equally as exhausting).
To give myself some credit, I believe that the first day was more so mentally taxing because it served as a double introduction for me. The obvious introduction was the description of the different techniques: Fluorescent Correlation Spectroscopy (FCS), Fluorescent Cross Correlation Spectroscopy (FCCS), Forster Resonance Energy Transfer (FRET), Fluorescent Lifetime Imaging (FLIM), and Fluorescent Recovery After Photobleaching (FRAP). The second introduction, which now I recognize as equally apparent is the biophysics behind all of these imaging techniques.
Accompanying every picture or description of technique there was some type of formula. Sometimes they were simple ratios that promoted a correlation in data, other times they were multivariable functions sporting integrals with sprinkles of sine, cosine, and tangent-clearly the demonic offspring of geometry and calculus. Really though-once I became familiar with what the different variables meant, they were not as intimidating.
Before this course my experience with microscopy has been that you use the images you obtain to verify the presence of a structure or event; you use microscopes to provide qualitative data. While this is still true (and an excellent use of microscopy), I have also learned that these techniques and many others that I have yet to encounter can be utilized to quantify processes in biological systems.
In this regard, these fluorescent techniques help to address a personal issue of mine with biology-generally speaking the underrepresentation of quantitative data as compared to the other hard sciences. Chemisty, physicist, engineers, and obviously mathematicians all take extensive courses in math, whereas biologists typically do not. Why?
At first, I thought the answer was simply that the other areas of science we more complicated and therefore inherently relied on mathematics for explanation. Conversely, biology is a simpler subject and could be evaluated effectively through qualitative observation.
However, now I prefer to think that biology is too complicated to be easily explained by mathematics. A living system has many variables whose effects all compound on one another to create the whole, therefore mathematics has shied away from the subject. Models and in vitro experiments simplify the system for experimental evaluation, but after everything I just said is biology really just the sum of its parts?
This week I learned that F-techniques can be used to quantify in vivo experiments and therefore can glean more accurate depictions of what is occurring in a living organism. For example, FRET (my personal favorite out of the bunch) and FCCS can be used to determine the dissociation constants, diffusion rates, and concentrations of proteins in biological pathways (this is an area of research interest for my PI- Dr. Sohail Ahmed).
On most days after the course, I would review my notes and create mini PowerPoints to further interact with the material. I will share them with you (once I have our microscope guru review it) so you can get a glimpse of what I have learned this week-enjoy!
I have also learned about Superresolution microscopy which can identify the location and movement of single molecules (proteins, lipids) in a system. One of the people who spoke on this topic is Dr. Boris Lukiyanchuck, whose research publication “Optical virtual imaging at 50 nm lateral resolution with a white-light nanoscope,” captured global attention. I found his personal website and am amazed by how many Nobel laureates he has worked with (he gives them thanks-honor thy mentors) and am impressed with how many places he has worked abroad (can I count him as an example for how working abroad makes you a successful person?). Here is his website if you are interested: http://www.lukiyanchuk.ru/
I am very thankful that Dr. Sohail Ahmed invited me to attend this course, even though it was meant for graduate students and Post-docs. It has opened my eyes to new research techniques and shown me how they can be applied to address the complexity of the life sciences.