Super resolution imaging made easy
Image: Open source method 'DeSOS' produces super-resolution images in live animals.
US-based researchers have unveiled an open source tool to view the physiological state of living organisms at the cellular level.
Developed by Professor Scott Howard and colleagues at the Indiana-based University of Notre Dame, 'DeSOS' combines conventional microscope images to create a single, high resolution, 3D image.
The imaging tool relies on two key methods, blind deconvolution (De), that can recover blurred images, and stepwise optical saturation super-resolution (SOS), which extends resolution beyond its typical diffraction limit.
As Howard points out: "Super-resolution approaches are limited... in longer-term intravital imaging, but this [technique] combines deconvolution with stepwise optical saturation microscopy to circumvent issues and image cells in their native physiological environment."
"Other than a traditional confocal or two-photon microscope, this approach requires no additional hardware," he adds.
Crucially, DeSOS can identify differences between the two uploaded images and produce one image with significantly greater clarity than previously possible with standard lab equipment.
Before and after with DeSOS: actin remodelling in zebrafish.
"Our program is also faster and has more functionality than a super-resolution microscope when evaluating living organisms,” claims Howard.
As part of the study, Professor Cody J. Smith from Biological Sciences, used DeSOS to improve imaging for his research on the development of spinal cords.
Smith and colleagues analysed specialised cells that grow and extend to form the nervous system of zebrafish.
According to the researchers, the images developed using DeSOS allowed them to see the structures of the zebrafish’s nervous system considerably better than traditional images taken on confocal and two-photon microscopes.
“Because of DeSOS, we could see individual neuronal fibres during the zebrafish spinal cord development process, including components of our research we couldn’t clearly recognize without this technology,” highlights Smith.
The researchers also used DeSOS in time-lapse imaging to generate super-resolution movies in zebrafish, creating a video of neuronal fibres growing to form branches during development, eventually extending towards the head and posterior to form the nervous system.
"The ease of acquisition and processing with this approach provides a universal technique for biologists to answer questions in living animals," says Howard.
Research is published in Development.