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Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging
Published on: December 12, 2012
A versatile platform for two-photon neuronal population voltage imaging across cortical depths
Jingkun Guo1, Kevin Barber1, M Agustina Frechou1
1Laboratory of Neurotechnology and Biophysics, The Rockefeller University, New York, NY, USA.
Nature Methods
|July 23, 2026
Summary
We developed FlatMux, a novel two-photon imaging system, to overcome limitations in voltage indicator imaging. This scalable platform enhances signal-to-noise ratio and reduces photobleaching for clearer neuronal activity recordings.
Area of Science:
- Neuroscience
- Optical Imaging
- Biotechnology
Background:
- Genetically encoded voltage indicators offer high spatiotemporal resolution for neuronal activity monitoring.
- Limitations include fast dynamics, low signal-to-noise ratio (SNR), photobleaching, and inefficient scaling to large neuronal populations.
- Suboptimal optical acquisition schemes hinder broader utility.
Purpose of the Study:
- To introduce a versatile, scalable, and efficient two-photon optical imaging system.
- To address the challenges of temporal dynamics, SNR, and photobleaching in voltage indicator imaging.
- To enable efficient scale-up for recording activity in larger neuronal populations.
Main Methods:
- Development of a flexible lateral-temporal multiplexing (FlatMux) platform.
- Implementation of a two-photon optical imaging system.
- Demonstration of reconfigurable modes including large field-of-view, high-speed (2-kHz), deep-tissue (500-µm), dual-plane, and high-SNR imaging.
Main Results:
- FlatMux platform demonstrated flexible reconfigurability for diverse recording needs.
- Achieved high-speed, deep-tissue, dual-plane, and high-SNR imaging with minimized pixel crosstalk and bleaching.
- Successfully recorded cortical spiking activity and subthreshold neuronal activity.
Conclusions:
- FlatMux provides a scalable and efficient solution for multiphoton voltage imaging in the mammalian cortex.
- The system overcomes key limitations of current voltage indicator technologies.
- Enables advanced studies of complex brain functions at single-spike and single-trial levels in large neuronal populations.
