One of the problems with
imaging living biological cells is that they don’t want to be held
still. Or, more accurately, they don’t want to be held to a surface like
a microscope slide. Prepping and fixing the cells changes them
irrevocably, altering whatever a scientist was trying to observe. Red
blood cells are like this: try to stick them fast to a cover slip and
their form changes, killing them before anyone gets quality images out
of the scope. Lots of bacteria are like this, too. But what if they
didn’t have to be stuck against a surface at all? What if they could be
held quite still without a physical touch?
“The principle underlying this laser beam is similar to the concept to be found in the television series Star Trek,” says Dr. Thomas Huser, head of the research group.
Continuing to build on the same underlying concepts as the
optical tweezers
from earlier this fall, physicists from the University of Bielefeld
have further developed that procedure for use in superresolution
fluorescence microscopy. The idea is that this new method can get
fluorescence images of living cells with resolution we could heretofore
only get with electron microscopy, which you can only use on nonliving
cells. Consequently, even living cells that resent being affixed to a
plate can now be imaged with higher resolution.
Distribution of the genetic material inside these rod-shaped E. coli bacteria. Image: Diekmann, Huser et al, 2016
To obtain images with such microscopes,
researchers add fluorescent probes to the cells they wish to study,
which then light up when a laser beam is directed at them. The
experimental setup then uses sensors to record this fluorescence, so
that researchers can even get three-dimensional images of the cells.
In their new method, the Bielefeld researchers use a
second laser beam as a single-cell optical trap — in other words, a
tiny tractor beam
— so that the cells float under the microscope and can be moved at
will. “When this laser beam is directed towards a cell, forces develop
within the cell that hold it within the focus of the beam,”
says
Robin Diekmann, coauthor. What forces? Depending on the power of the
laser, the tractor beam has a quality called “stiffness,” against which
cells experience a restoring force.
Using their new method, the physicists
succeeded in holding and rotating first a polystyrene bead and then
individual bacterial cells in midair, on a plane just a few micrometers
above the slide, in such a way that they could take images of the cells
from several sides and focus the image throughout the whole depth of the
cells. With the ability to rotate the cells in 3-space, the researchers
can study the three-dimensional structure of the cells’ genetic
material, at micron resolutions.
Abbreviations:
AOTF: acousto-optical tunable filter, FT: focusing telescope, TM:
translatable mirror, DCM: dichroic mirror, NA: numerical aperture, TIR:
total internal reflection, ND: neutral density filter wheel, PMF:
polarization-maintaining fiber, 4f-T: 4f-telescope, M: mirror, BP:
band-pass filter, SP: short-pass filter. Image and description:
Diekmann, Huser et al, 2016
Diekmann and colleagues’ next steps will be to
deploy the combination of fluorescence microscopy and optical tweezers
to study cells through different parts of their life cycles, including
watching bacteria and other cells as they’re infected by other
pathogens. No word on how long it’ll be before we have a tractor beam
that can scale up for starships.
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