To capture the cellular images, the interdisciplinary UB team of biologists, chemists and physicists, led by Prasad, utilized an advanced biophotonic approach that combines three techniques: a nonlinear, optical imaging system (CARS or Coherent anti-Stokes Raman scattering), TPEF (two-photon excited fluorescence), which images living tissue and cells at deep penetration and Fluorescence Recovery after Photobleaching to measure dynamics of proteins.
"For the first time, this approach allows us to monitor in a single scan, four different types of images, characterizing the distribution of proteins, DNA, RNA and lipids in the cell," says Aliaksandr V. Kachynski, PhD, research associate professor at the ILPB and co-author.
The resulting composite image integrates in one picture the information on all four types of biomolecules, with each type of molecule represented by a different color: proteins in red, RNA in green, DNA in blue and lipids in grey, as shown on the PNAS cover.
Multiplex imaging provided new information on the rate at which proteins diffuse through the cell nucleus, the UB scientists say.
Before apoptosis was induced, the distribution of proteins was relatively uniform, but once apoptosis develops, nuclear structures disintegrate, the proteins become irregularly distributed and their diffusion rate slows down, says Artem Pliss, PhD, research assistant professor at the ILPB and co-author on the paper.
"This research gives us the unique ability to study and improve our understanding of individual subcellular structures and the transformations they go through," says Pliss.
Such precise information will be especially useful for monitoring how specific cancer drugs affect individual cells.
"For example, say drug therapy is being administered to a cancer patient; this system will allow for the monitoring of cellular changes throughout the treatment pro
|Contact: Ellen Goldbaum|
University at Buffalo