Light Beads Microscopy: A Breakthrough in Volumetric in vivo Brain Imaging
In their recent publication, Jeffrey Demas and co-authors introduced “Light Beads Microscopy”, an important technological breakthrough in 2 photon microscopy. The
The Light Beads Microscopy (LBM) unit is a groundbreaking add-on for a two-photon microscope, enabling the acquisition of a single volume with a depth of approximately 0.5mm at the same speed as capturing a single plane in a conventional microscope. This innovative module was developed by Dr. Alipasha Vaziri’s Laboratory of Neurotechnology and Biophysics at Rockefeller University in New York, NY.
Current two-photon scanning microscopy techniques for brain imaging often face challenges in balancing image resolution, acquisition speed, and signal-to-noise ratio (SNR). These challenges become particularly pronounced during volumetric imaging due to the light-scattering properties of brain tissue.
Light Beads Microscopy (LBM) offers a solution by optimizing spatiotemporal signal acquisition to record data at rates limited by the fluorescence lifetime of neural calcium indicators, such as GCaMP. LBM leverages high-energy femtosecond laser pulses at a relatively low repetition rate (~4.5 MHz to 5 MHz) to enable the highly temporally multiplexed acquisition of up to 30 axial planes within the same timeframe required for traditional two-photon microscopy to capture a single plane. Additionally, LBM enhances SNR by utilizing one excitation pulse per voxel.
Laser Frequency (MHz) | Maximum Number of Planes* |
4.5 | 32 |
4.7 | 30 |
5 | 28 |
*assuming a temporal window of 6.8ns for the demultiplexing channels
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In their recent publication, Jeffrey Demas and co-authors introduced “Light Beads Microscopy”, an important technological breakthrough in 2 photon microscopy. The
MBF’s software utility is underscored by the number of references it receives in the worlds most important scientific publications.
Bellafard, G. Namvar, J. Ko, A. Vaziri, and P. Golshani.
Volatile working memory representations crystallize with practice.View Publication
J. Manley, S. Lu, K. Barber, J. Demas, H. Kim, D. Meyer, F. Martínez Traub, and A. Vaziri.
Simultaneous, cortex-wide dynamics of up to 1 million neurons reveal unbounded scaling of dimensionality with neuron number.View Publication
Demas, J., Manley, J., Tejera, F. et al.
High-speed, cortex-wide volumetric recording of neuroactivity at cellular resolution using light beads microscopyView Publication
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