New software helps in research
I don’t remember exactly when I got my first microscope. It could have been my birthday, or maybe Christmas, but I remember sitting at the dining room table on a sunny day trying to catch the light just right in the mirror at the base to illuminate the magnified image.
And I remember the excitement of getting a new one years later with an electric bulb, so I could peer into the mysteries that lay hidden in smallness – even on cloudy days.
Using those simple light microscopes at home and later in school during biology classes, I was able to see just some of what is invisible to the naked eye.
Immunofluorescence (IF) microscopy is an imaging technique used by researchers to learn about the location and quantity of a particular protein in a sample. A sample can be a small piece of muscle, skin or other tissue taken from a biopsy. And the pictures made by IF microscopy are beautiful! They even have art competitions for the images produced using IF microscopy.
If a researcher wants to know how much dystrophin (an important protein for muscle function) is located in the muscle fibers they’re examining, they could use IF microscopy to find out.
They would use antibodies chemically labeled with dyes that light up (fluoresce) under the right conditions. The antibodies ensure that the dyes only stick to the proteins you want to see.
In itself, this is pretty amazing stuff. But it gets even better.
Dystrophin, the protein in our example, is the key element that is missing or defective in people with Duchenne or Becker muscular dystrophies. Understanding how much dystrophin is in the muscles of children with these forms of muscular dystrophy, and how the dystrophin level relates to muscle function, is critical for developing new therapies and understanding what makes a potential therapy effective.
Using immunofluorescence microscopy to illuminate the dystrophin in the cells is the first step, but developing an automated, reliable, unbiased approach to quantifying it can be challenging. In a study with hundreds of images to be quantified, it would take one person a long time to do this. And because of human variability in interpreting the images, having multiple people could introduce inconsistencies and make it hard to rely on the results.
Enter Tatyana Vetter, PHD, research scientist and director of microscopic imaging in the Center for Gene Therapy at Nationwide Children’s Hospital. She has developed and published an approach that does just that. By developing a program for microscopy software that is commonly used and easily accessible, she has built an approach that provides consistent results with little training time.
Dr. Vetter’s approach is a series of many steps that are combined to create what the software calls “recipes,” allowing users to apply the steps automatically without needing to do any coding or complicated technical adjustments to get the measurements they need for their studies.
And she and her collaborators have decided to share it freely with other researchers.
The approach of using recipes in the system, in the style of a gift that keeps on giving, is modifiable to measure many proteins on many tissues, creating a multitude of possible applications.
Abbie Roth is managing editor of Pediatrics Nationwide and Science Communication at Nationwide Children's Hospital.
Reach Roth at Abbie.roth@nationwidechildrens.org