In vivo imaging of deep cortical layers using a microprism

Area of Science:

• Neuroscience
• Biomedical Engineering
• Optical Imaging

Background:

• In vivo imaging of deep cortical layers is challenging.
• Existing methods often lack the necessary resolution or field-of-view.
• Visualizing neuronal morphology and microvasculature in deep brain regions requires advanced techniques.

Purpose of the Study:

• To introduce a novel microprism probe for in vivo imaging of the cerebral cortex.
• To demonstrate the capability of the microprism probe for visualizing deep cortical layers and cellular structures.
• To detail the protocol, surgical procedure, and microscope setup for using the microprism probe.

Main Methods:

• Development of a 1-mm microprism probe with a reflective coating for internal light reflection.
• Simultaneous imaging of multiple cortical layers with a large field-of-view (approx. 900 microm).
• Non-survival surgical procedures and specific microscope configurations were employed.

Main Results:

• High-resolution in vivo imaging of layer V pyramidal neurons, including apical dendrites and dendritic spines.
• Visualization of the cortical vasculature network using fluorescent dye injection.
• Measurement of red blood cell (RBC) velocity and flux rates in capillaries via line-scanning.

Conclusions:

• The microprism probe offers an effective method for in vivo visualization of deep cellular structures in the cortex.
• This technique allows for the study of neuronal morphology and microvascular dynamics with unprecedented detail.
• The microprism probe represents a significant advancement in neuroimaging capabilities.