Fluorescent microscopy adds a new dimension with Rice University lab's invention
Rice University researchers have added a new dimension to their breakthrough technique that expands the capabilities of standard laboratory microscopes.
Rice University researchers have created a method to design custom masks that transform 2D fluorescent microscopy images into 3D movies.
Credit: Landes Research Group/Rice University
An electron microscope image shows a phase mask programmed by an algorithm created at Rice University. The customizable mask lets the Rice lab take images of molecules to determine their depth and how fast they move, even if they are faster than a single camera frame.
Credit: Landes Research Group/Rice University
Two years ago, the Rice lab of chemist Christy Landes introduced super temporal resolution microscopy, a technique that allowed researchers to image fluorescent molecules 20 times faster than traditional lab cameras normally allow.
They've now developed a general method to let a microscope capture 3D spatial information along with the fourth dimension, molecular movement over time.
This, they say, will help scientists who study dynamic processes view where molecules of interest are located and how fast they move -- for example, within living cells.
The Rice method to expand the capabilities of existing wide-field fluorescence microscopes is detailed in the team's open-access paper in Optics Express.
It describes the creation of custom phase masks: transparent, spinning disks that manipulate light's phase to change the shape of the image captured by the microscope's camera. The shape contains information about a molecule's 3D position in space and how it behaves over time within the camera's field of view.
A phase mask turns what seems like an inconvenience, the blurry blob in a microscope image, into an asset. Scientists give this blob a name -- point spread function -- and use it to get details about objects below the diffraction limit that are smaller than all visible light microscopes are able to see.
The original work used a rotating phase mask that transformed light from a single fluorescent molecule into what the researchers called a rotating double helix. The captured image appeared on the camera as two glowing disks, like the lobes of a barbell.
In the new work, the rotating barbells let them see not only where molecules were in three-dimensional space, but also gave each molecule a time stamp.
The heart of the new work lies in algorithms by lead author and Rice electrical and computer engineering alumnus Wenxiao Wang. The algorithms make it practical to design custom phase masks that modify the shape of the point spread function.
"With the double helix phase mask, the time information and spatial information were connected," said co-author Chayan Dutta, a postdoctoral researcher in Landes' lab. "The lobes' rotation could express either 3D space or fast time information, and there was no way to tell the difference between time and space."
Better phase masks solve that problem, he said. "The new phase mask design, which we call a stretching lobe phase mask, decouples space and time," Dutta said. "When the targets are at different depths, the lobes stretch farther apart or come closer, and the time information is now encoded just in the rotation."
The trick is to manipulate light at the spinning phase mask to optimize the pattern for different depths. That is accomplished by the refractive pattern programmed into the mask by the algorithm. "Each layer is optimized in the algorithm for different detection depths," said graduate student and co-author Nicholas Moringo. "Where before, we could see objects in two dimensions over time, now we can see all three spatial dimensions and fast time behavior simultaneously."
"Wide-field fluorescence microscopes are used in many fields, especially cell biology and medical imaging," Landes said. "We are just starting to demonstrate how manipulating light's phase within a microscope is a reasonably simple way to improve space and time resolution compared to developing new fluorescent tags or engineering new hardware improvements."
One important outcome that could have broad appeal, she said, is that the researchers generalized the phase mask design so researchers can fabricate masks to create virtually any arbitrary pattern.
To demonstrate, the group designed and fabricated a mask to create a complex point spread function that spells out RICE at different focal depths. A video shows the ghostly letters appear and disappear as the microscope moves to different depths above and below the focal plane.
Such flexibility will be useful for applications like analyzing processes inside living cancer cells, a project the lab hopes to pursue soon with Texas Medical Center partners.
"If you have a cell on a glass slide, you'll be able to understand where objects in the cell are in relationship to each other and how fast they move," Moringo said. "Cameras aren't fast enough to capture all of what happens in a cell, but our system can."
Co-authors are Rice graduate student Fan Ye; former Rice postdoctoral researcher Hao Shen, now an assistant professor at Kent State University; and Jacob Robinson, a Rice assistant professor of electrical and computer engineering. Wang is now a software engineer at Google. Landes is a professor of chemistry and of electrical and computer engineering.
The National Science Foundation and the Welch Foundation supported the research.
Read the paper at https:/
This news release can be found online at https:/
Follow Rice News and Media Relations via Twitter @RiceUNews.
Related materials: Molecular imaging hack makes cameras 'faster': http://news.
Landes Research Group: https:/
Wiess School of Natural Sciences: https:/
George R. Brown School of Engineering: https:/
Rice University researchers have created an algorithm to produce custom phase masks that help analyze molecular processes below the diffraction limit and 20 times faster than traditional cameras. The team produced a mask that shows the depth of an object by spelling out RICE as the focal plane changes. (Credit: Wenxiao Wang/Landes Research Group)
Images for download:
Rice University researchers have created a method to design custom masks that transform 2D fluorescent microscopy images into 3D movies. (Credit: Landes Research Group/Rice University)
An electron microscope image shows a phase mask programmed by an algorithm created at Rice University. The customizable mask lets the Rice lab take images of molecules to determine their depth and how fast they move, even if they are faster than a single camera frame. (Credit: Landes Research Group/Rice University)
The point spread functions of single molecules, captured as double lobes through a phase mask (left), can tell researchers where the molecule is in 3D space. The distance between the lobes gives them the molecule's depth. (Credit: Landes Research Group/Rice University)
Rice University postdoctoral researcher Chayan Dutta sets up a microscope for new experiments with super temporal resolution microscopy, a technique the lab has advanced with the ability to make custom phase masks. (Credit: Jeff Fitlow/Rice University)
Located on a 300-acre forested campus in Houston, Rice University is consistently ranked among the nation's top 20 universities by U.S. News & World Report. Rice has highly respected schools of Architecture, Business, Continuing Studies, Engineering, Humanities, Music, Natural Sciences and Social Sciences and is home to the Baker Institute for Public Policy. With 3,962 undergraduates and 3,027 graduate students, Rice's undergraduate student-to-faculty ratio is just under 6-to-1. Its residential college system builds close-knit communities and lifelong friendships, just one reason why Rice is ranked No. 1 for lots of race/class interaction and No. 2 for quality of life by the Princeton Review. Rice is also rated as a best value among private universities by Kiplinger's Personal Finance. To read "What they're saying about Rice," go to http://tinyurl.
Jeff Falk 713-348-6775
Mike Williams 713-348-6728
Jeff Falk | EurekAlert!
Spintronics: Researchers show how to make non-magnetic materials magnetic
06.08.2020 | Martin-Luther-Universität Halle-Wittenberg
Manifestation of quantum distance in flat band materials
05.08.2020 | Institute for Basic Science
Scientists at the Fraunhofer Institute for Laser Technology ILT have come up with a striking new addition to contact stamping technologies in the ERDF research project ScanCut. In collaboration with industry partners from North Rhine-Westphalia, the Aachen-based team of researchers developed a hybrid manufacturing process for the laser cutting of thin-walled metal strips. This new process makes it possible to fabricate even the tiniest details of contact parts in an eco-friendly, high-precision and efficient manner.
Plug connectors are tiny and, at first glance, unremarkable – yet modern vehicles would be unable to function without them. Several thousand plug connectors...
An international research team has found a new approach that may be able to reduce bone loss in osteoporosis and maintain bone health.
Osteoporosis is the most common age-related bone disease which affects hundreds of millions of individuals worldwide. It is estimated that one in three women...
Traditional single-cell sequencing methods help to reveal insights about cellular differences and functions - but they do this with static snapshots only...
“Core-shell” clusters pave the way for new efficient nanomaterials that make catalysts, magnetic and laser sensors or measuring devices for detecting electromagnetic radiation more efficient.
Whether in innovative high-tech materials, more powerful computer chips, pharmaceuticals or in the field of renewable energies, nanoparticles – smallest...
An international research team with Prof. Cornelia Denz from the Institute of Applied Physics at the University of Münster develop for the first time light fields using caustics that do not change during propagation. With the new method, the physicists cleverly exploit light structures that can be seen in rainbows or when light is transmitted through drinking glasses.
Modern applications as high resolution microsopy or micro- or nanoscale material processing require customized laser beams that do not change during...
23.07.2020 | Event News
21.07.2020 | Event News
07.07.2020 | Event News
06.08.2020 | Earth Sciences
06.08.2020 | Power and Electrical Engineering
06.08.2020 | Life Sciences