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Updated: Jun 22, 2026

In vivo Imaging of Deep Cortical Layers using a Microprism

In vivo Imaging of Deep Cortical Layers using a Microprism

Published on: August 27, 2009

Microprisms for in vivo multilayer cortical imaging.

Thomas H Chia¹, Michael J Levene

¹Department of Biomedical Engineering, Yale University, New Haven, Connecticut, USA.

Journal of Neurophysiology

|June 5, 2009

Summary

Microprisms enable in vivo two-photon imaging of multiple neocortical layers, overcoming depth limitations. This technique provides high-resolution views of neuronal structures and blood flow, comparable to slice preparations with minimal tissue damage.

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Last Updated: Jun 22, 2026

In vivo Imaging of Deep Cortical Layers using a Microprism

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Published on: August 27, 2009

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Long-Term Imaging of Identified Neural Populations using Microprisms in Freely Moving and Head-Fixed Animals

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Published on: January 19, 2024

Area of Science:

Neuroscience
Optical Imaging
Microscopy

Background:

Cortical slices offer multi-layer imaging but lack native physiological context.
In vivo two-photon imaging is limited in depth (<500 microm) and struggles with simultaneous multi-layer imaging due to cortical stratification.

Purpose of the Study:

To introduce and validate the use of 1-mm microprisms for in vivo, two-photon neocortical imaging.
To enable simultaneous imaging of multiple cortical layers, including layer V, at an angle similar to slice preparations.

Main Methods:

Utilized 1-mm microprisms for in vivo, two-photon imaging of the mouse motor and somatosensory cortex.
Collected images with a nearly 900-microm field of view and high magnification, resolving dendritic spines.
Performed functional imaging of blood flow and assessed tissue viability using H&E, propidium iodide, and Nissl staining.

Main Results:

Microprism imaging achieved simultaneous visualization of multiple cortical layers, including layer V, with high resolution.
Images were comparable to traditional slice preparations, capturing dendritic spines and enabling functional blood flow analysis.
Histological analysis confirmed native tissue stratification and limited neuronal damage (<100 microm) at the tissue-glass interface.

Conclusions:

1-mm microprisms are a straightforward tool for advanced in vivo neocortical imaging.
This technique overcomes previous depth and stratification limitations, offering significant advantages for neuroscience research.
Microprisms facilitate high-resolution imaging of neuronal structures and physiological processes within the intact cortex.