Multicolor two-photon light-patterning microscope exploiting the spatio-temporal properties of a fiber bundle.

Area of Science:

• Neuroscience
• Optical Engineering
• Biotechnology

Background:

• Genetically encoded tools enable light-based control of neuronal activity.
• All-optical methods offer precise, single-cell resolution targeting in 3D.
• Existing techniques are complex and often limited to a single wavelength.

Purpose of the Study:

• To develop a simpler, dual-color optical approach for precise 3D neural targeting.
• To overcome the limitations of single-wavelength and complex existing methods.
• To enable reconfigurable targeting of large neuronal populations.

Main Methods:

• Utilized a fiber bundle and a spatial light modulator for light patterning.
• Employed two-photon excitation for high-resolution spot generation.
• Leveraged fiber bundle properties for simultaneous dual-color patterning in 3D.

Main Results:

• Demonstrated high-resolution excitation spot generation in a 3D volume.
• Achieved simultaneous dual-color two-photon light patterning.
• Showcased a simple and effective method for multicolor cell targeting.

Conclusions:

• The novel optical approach offers a simple and suitable alternative for precise multicolor cell targeting.
• This technique simplifies the setup and operation of advanced optogenetic tools.
• Facilitates precise manipulation of neuronal activity in living tissues with multiple colors.