Researchers at Texas A&M University have developed a new microscopy technique that addresses a long-standing limitation in biological imaging: the trade-off between viewing fine cellular detail and looking deep inside living tissue. The method, known as 2PE-ISM-SOFI, combines two established techniques to improve resolution without sacrificing imaging depth.
The team, led by Aart Verhoef, assistant professor of biophotonics in the Department of Soil and Crop Sciences and joint professor in the Department of Physics and Astronomy, published their findings in the Journal of Biomedical Optics. The new technique achieves lateral resolution down to approximately 75 nanometers, which is roughly five times sharper than conventional two-photon microscopy.
This allows for the visualization of structures smaller than 100 nanometers, a scale roughly a thousand times thinner than a human hair, within intact samples.
Conventional two-photon microscopy is widely used because it can peer deep into biological samples with minimal damage, making it suitable for studying living tissue in fields such as neuroscience, developmental biology, and plant sciences. However, this depth has typically required staying near the sample’s surface to see fine detail.
Super-resolution techniques like STED and PALM can resolve features below 100 nanometers but usually only close to the surface. The new 2PE-ISM-SOFI method bridges this gap by maintaining the depth advantage of two-photon imaging while achieving higher resolution.
Verhoef stated, “This technique builds on two microscopy techniques that were limited by depth and detail, but this technique allows us to look deep into tissue and improves the resolution.”
