Researchers have developed a new fluorescence microscopy approach that — by acquiring three views of a sample simultaneously — can give them a better understanding of how healthy cells work and what goes wrong when diseases occur.
Biologists usually use fluorescence microscopy to study things like embryo development and the processes within cells. But most of the methods that they use don’t capture much of the fluorescence emitted for the sample, which means a loss of information and poor image resolution.
The new "multi-view" approach is an improvement of an older technique that the researchers developed called dual-view plane illumination microscopy (diSPIM), which is used by scientists around the world. It leverages a thin sheet of light and two objective lenses to excite and detect fluorescence.
The resolution of diSPIM was restricted by the numerical aperture of the upper lenses and the fluorescence that’s emitted, but not captured. The researchers reasoned that if they could image the neglected signal by adding a higher numerical aperture lens that acquired the bottom view, then they could increase the lateral resolution.
They tested the multi-view technique by imaging biological samples and were able to see detailed features that are not usually observable. Using the light-sheet mode, they were able to clearly see the 3-D dynamic nature of small protrusions on living white blood cells when they captured 150 triple-view images over the span of 40 minutes.
The team is now researching more biological applications for the new system, and is also working to extend the technique to other microscope modalities like confocal microscopy.
