Two-photon Micropuncture

Two-photon micropuncture is a live-tissue technique that combines two-photon microscopy with a fine micropipette to visualize and access cells, vessels, or tissue compartments deep within optically scattering specimens. Near-infrared femtosecond laser pulses excite fluorophores only at the focal point, while real-time imaging guides the micropipette to a selected target for sampling, injection, or localized manipulation with limited photodamage outside the focal plane. In cancer research, this approach can examine tumor vasculature and the tumor microenvironment, measure or alter local conditions, and relate microscopic structure to functional responses, supporting studies of drug delivery, vascular permeability, tumor cell behavior, and treatment effects in living models.

Two-photon Micropuncture - Related Videos

Research

JoVE Journal - Biology
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Micropuncture of Bowman's Space in Mice Facilitated by 2 Photon Microscopy

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Cited by 5 •

2018

We present use of 2-photon microscopy to place a micropipette within Bowman's urinary space in mice, combining 2 foundational techniques of renal physiology. Use of 2-photon microscopy overcomes critical limitations of conventional microscopy for micropuncture renal physiology studies.

Research

JoVE EoE - Urinary Tract Cancer

Fluid Aspiration from Mouse Glomeruli: A Method to Aspirate Glomerular Fluid from Mouse Using Two-Photon Mediated Micropuncture

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2023

This video showcases fluid aspiration from mouse glomeruli using the two-photon mediated micropuncture technique. The technique provides access to the living mouse’s glomeruli and thus helps gain insights into renal physiology.

Two-Photon In Vivo Imaging of the Mouse Retina

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2025

Source: Wang, Z., et al. Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina. J. Vis. Exp. (2021).This video demonstrates a technique for in vivo retinal imaging using two-photon microscopy. Anesthetized mice are prepared for imaging by securing their heads, Applying eye lubricant, and placing a coverslip over the eye. Retinal ganglion cells (RGCs) expressing fluorescent proteins are first visualized using epifluorescence for alignment. Finally, two-photon imaging is employed,...

Fabrication of 1-D Photonic Crystal Cavity on a Nanofiber Using Femtosecond Laser-induced Ablation

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Cited by 3 •

2017

We present a protocol for fabricating 1-D photonic crystal cavities on subwavelength diameter silica fibers (optical nanofibers) using femtosecond laser-induced ablation.

Fabrication and Testing of Photonic Thermometers

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Cited by 1 •

2018

We describe the process of fabrication and testing of photonic thermometers.

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