Non-invasive, live cell imaging is critical to modern biological and medical research. Two-photon fluorescence microscopy is an important method used to visualize tissue in depth. However, it suffers from constraints on image contrast because of out-of-focus fluorescence that overwhelms the detected signal. This technology proposes a method to extend the depth limit and increase contrast by performing stimulated emission reduced fluorescence microscopy (SERF). The subsequent reconstructed image using this method has a significantly improved signal-to-background contrast. This allows for improved depth and resolution of two-photon fluorescence microscopy for live cell imaging of biological specimens including brain slices, embryos, whole organs, and even live animals.
This technology uses a focused laser beam that preferentially switches fluorescence on and off at the focal point. This effect selectively induces stimulated emission, reducing the fluorescence intensity at the focal point, but does not decrease fluorescence of the background. This leads to a large contrast between the image and the background, which is significantly improved over other two-photon microscopy methods. Additionally, this method is also beneficial in that it increases the imaging depth limit.
The technology has been demonstrated by both analytical theory and numerical simulations.
Patent Pending (WO/2013/173698)
Patent Pending (US 20150168703)
Tech Ventures Reference: IR CU12311