New Fluorescence Microscopy Technique for Nanostructure Visualization Within Cells

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Researchers at Purdue University have developed a new technique to dramatically enhance the resolution achievable when imaging intracellular structures with super-resolution fluorescence microscopy. The technique uses the distortions created by a specimen to pinpoint the location of individual molecules, and thereby infer the location of intracellular structures. The technique could be particularly useful in studying neurological diseases such as Alzheimer’s.

Understanding
the structural complexity of cells involved in disease is crucial to developing
treatments. However, at the intracellular level, it has been difficult to
accurately visualize small structures using fluorescence microscopy because of
the distortion caused by light emitted by molecules within the specimen.

Now,
researchers at Purdue have turned this situation on its head, and are using the
distortion produced by molecules within the specimen to plot their location,
enabling significantly increased resolution.     

“Our technology allows us to measure wavefront distortions induced by the specimen, either a cell or a tissue, directly from the signals generated by single molecules – tiny light sources attached to the cellular structures of interest,” said Fang Huang, a researcher involved in the study published in Nature Methods. “By knowing the distortion induced, we can pinpoint the positions of individual molecules at high precision and accuracy. We obtain thousands to millions of coordinates of individual molecules within a cell or tissue volume and use these coordinates to reveal the nanoscale architectures of specimen constituents.”

The
researchers can pinpoint the location of specific biomolecules in cells and
tissues to within a few nanometers, allowing for incredibly accurate
visualization of tiny structures. The technology could be useful in studying pathological
processes within cells, paving the way for new treatments.

“This
technical advancement is startling and will fundamentally change the precision
with which we evaluate the pathological features of Alzheimer’s disease,” said Gary
Landreth, another researcher involved in the study. “We are able to see smaller
and smaller objects and their interactions with each other, which helps reveal
structure complexities we have not appreciated before.”

Here’s a video of a 3D animation generated using the new technique:

Study in Nature Methods: Three-dimensional
nanoscopy of whole cells and tissues with in situ point spread function
retrieval

Via: Purdue





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