A pioneering 3D imaging method has been applied to heart cells
for the first time, allowing researchers to trace waves that can
cause arrhythmia.
Rising levels of calcium in heart muscle cells cause contraction
of the muscle, helping to regulate the beating of the heart. The
rise in calcium levels is usually uniform, but sometimes there is a
spontaneous release of calcium from isolated regions of the cells,
creating a wave of calcium.
These waves can cause arrhythmia, the irregular beating of the
heart. Arrhythmia accounts for approximately 50 percent of deaths
in patients with heart failure.
However, how and why calcium waves originate has been difficult
to study with conventional microscopy techniques. Now, physicists
from Imperial College London have collaborated with scientists from
Imperial's National Heart and Lung Institute (NHLI) to shine a new
kind of light on the problem.
Their results reveal initial findings that calcium waves
originate from healthy parts of heart muscle cells, and not
degraded regions as the researchers had expected.
The technique, called oblique plane microscopy (OPM), was
invented by physicists in Imperial's Photonics group. In order to
study cells in 3D, scientists most commonly use confocal
microscopy, which looks at one point on the sample at a time.
OPM looks at a layer of the sample at a time instead of a point,
and combines this with a method to rapidly sweep the layer being
imaged through the specimen.
This allows video-rate 3D imaging of features at the
sub-cellular scale. This is particularly important for looking at
calcium waves, since they are rare events and their point of origin
within the cell is not known before it happens.
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Using this technique, researchers at the NHLI investigated
single heart muscle cells isolated from a rat model of heart
failure.
Within heart muscle cells, there are structures in the cell
membrane called transverse tubules or t-tubules, which are
essential for normal calcium release. In patients with heart
failure, the structure of t-tubules is degraded.
The researchers had speculated that these faulty structures were
the origin of calcium waves, but when they looked at the microscopy
data, the opposite pattern emerged.
Calcium waves were more frequent from regions of the cell where
the normal t-tubule structure was preserved. "We thought more
calcium waves would be produced from regions of deranged t-tubules,
but we were surprised to find the opposite appears to be true,"
said Ken MacLeod of Imperial.
"However, this was only a small-scale study to test the
technique. We still expect the derangement of t-tubules plays a
role in the poor function of failing cells, and hopefully with more
research we should be able to see what's going on in greater
detail. Knowing more about the origin of calcium waves would be an
important first step in combatting arrhythmia."
Chris Dunsby, also of Imperial, said: "Now we have proven OPM
can give real insights into these processes, we hope to improve the
technique and continue working with the NHLI.
The paper, High speed sCMOS-based oblique plane microscopy applied to
the study of calcium dynamics in cardiac myocytes, is published
in the Journal of Biophotonics.