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The neural activity of one million neurons in the mouse brain with unprecedented resolution
Capturing the intricate brain activity requires resolution, scale and speed-when millions of neurons actively send out signals from remote corners of the cerebral cortex in less than a second, we can distinguish with extreme clarity Rate to see them
.
Now, researchers have developed a microscope technology that allows scientists to accomplish this feat, capturing detailed images of a large number of cell activities at different depths of the brain with unprecedented speed and clarity
.
The research published in the journal “Nature Methods” demonstrates the innovative power of this kind of light-bead microscope.
"To understand the nature of the brain's tightly connected network, it is necessary to develop new imaging techniques that can capture the activity of neurons in a wide range of brain regions at high speed and single-cell resolution," said Rockefeller's Alipasha Vaziri
.
"Light bead microscopy will allow us to study biological problems in a way that was impossible before
Focusing microscope
Whether it is looking for a dangerous beard by swinging back and forth, or helping humans to hit a baseball's hand-eye coordination, animals rely on the brain's sense, movement, and visual areas of the call and response
.
Cells from the distant part of the cerebral cortex coordinate this feat through a network of neural activity that weaves the distant areas of the brain into an interconnected symphony
With the help of cutting-edge microscope technology, scientists are just now beginning to unravel this net
.
The combination of two-photon scanning microscopy and fluorescent labels is the gold standard for imaging neuronal activity in opaque brain tissue, which easily scatters light
But the two-photon microscope has a basic limitation
.
Neurobiologists need to record the simultaneous interaction between sensory, motor, and visual areas of the brain, but it is difficult to capture activity in such a wide area of the brain without sacrificing resolution and speed
Designing an ideal microscope to visualize the interactions between the various areas of the brain feels like blocking a hole in a shipwreck
.
In order to obtain high resolution, scientists often have to sacrifice scale-or shrink the image to obtain a larger structure, at the expense of resolution
Vaziri said: "We need to capture many neurons in the remote part of the brain at the same time with high resolution
.
" "These parameters are almost mutually exclusive
An innovative solution
The light bead microscope provides a creative solution and pushes the limit of imaging speed to the maximum achievable-only limited by the physical properties of fluorescence itself
.
This is done by eliminating the "dead time" between consecutive laser pulses, when no neural activity is recorded and scanning is required at the same time
This technique involves decomposing a strong pulse into 30 smaller sub-pulses—each of which has a different intensity—these sub-pulses sneak into the brains of scattered mice at 30 different depths, but induce the same number at each depth.
Fluorescence
.
This is achieved through a mirror cavity, which ignites each pulse staggeredly and ensures that they can reach the target depth through a microscope focusing lens
.
With this method, the only limit to the sample recording rate is the time for the fluorescent label to glow
.
This means that a large number of brain regions can be recorded at the same time, while a traditional two-photon microscope can only capture a small number of brain cells
.
Vaziri and colleagues then integrated the lamp bead microscope test into an optical microscope platform that allowed access to a large brain volume to record the activity of more than one million neurons in the cortex of the entire mouse brain for the first time
.
Since Waziri's method is an innovation based on two-photon microscopy, many laboratories already have or can commercially obtain the technology necessary to implement light bead microscopy, as described in the paper
.
Laboratories less familiar with these technologies can benefit from the more widely used simplified, stand-alone modules that Vaziri is currently developing
.
"We have no good reason not to do this five years ago," he said
.
"This was possible because microscopy and laser technology already existed
.
No one thought of this
.
"
Ultimately, our goal is to supplement, not replace, existing technologies
.
"For some neurobiological problems, a standard two-photon microscope is sufficient," Vaziri said
.
But the light bead microscope allows us to solve problems that cannot be solved by existing methods
.
"
DOI
10.
1038 / s41592-021-01239-8
Article title
High-speed, cortex-wide volumetric recording of neuroactivity at cellular resolution using light beads microscopy
