The new development comes from the U.S. National Institute of Biomedical Imaging and Bioengineering. For this, scientists have successfully combined two different microscope technologies together in order to produce sharper images which show the rapidly moving processes inside a living cell.
The research builds upon the microscopy method of total internal reflection fluorescence. This method allows for a thin region of a specimen, usually less than 200 nanometers to be observed. This process functions through illuminating a sample via a sharp angle so that the light reflects back. In doing so, this technique illuminates only the thin section of the sample that is located close to the coverslip. As the background light is reduced, an image of a far greater contrast image is produced.
There is a downside to total internal reflection fluorescence microscopy, however. This relates to how well the technique can capture tiny features within cells. While this can be accomplished gradually, the process is very slow. This decreases the usefulness of the method for examining living cells. This means many key cellular processes are invariably missed.
A new approach looks likely to change this. The alternate process comes involves the modification of a high-speed, super-resolution microscope to act like a total internal reflection fluorescence microscope. This process is described as instant structured illumination microscopy.
The process enables biologists to capture images of rapidly moving objects, as occurs with cell processes, some 10-times faster than other microscopes at a similar resolution. Such research will assist with application like cancer detection.
Discussing this breakthrough with the website Bioscience Technology, lead researcher Hari Shroff explains further: “Our method improves the spatial resolution of TIRF microscopy without compromising speed—something that no other microscope can do. We hope it helps us clarify high-speed biology that might otherwise be hidden or blurred by other microscopes so that we can better understand how biological processes work.”
The research has been published in the journal Nature Methods. The paper is titled “Single-shot super-resolution total internal reflection fluorescence microscopy.”
