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Cambridge Advanced Imaging Centre

 

Using our 2-photon scanning microscope for accurate 3D ablation

Two-photon imaging requires the use of high powered and pulsed lasers in order to achieve sufficient signal of the two-photon absorption process. These lasers have the added benefit of being able to achieve high localised energy densities that can ablate tissue.

Our microscope is equipped with a fast power modulation unit that allows specified regions of the scanned image plane to be treated with sufficient energy density to ablate and cut through tissue.

So far researchers in Cambridge have used this capability to:

Make accurate cuts (300nm in width and micrometres in length) to actomyosin cortex in Drosophila embryos

Shown here is labeled actomyosin cortex in a stage 9 Drosophila embryo. In this time-series a pulsed near-infrared laser is used for targeted ablation. Such experiments allow local measurements of tissue tension, essential for studying developmental mechanics. This experiment was part of research conducted by the Sanson Lab.

Trigger single cell death deep in Zebrafish retina in order to observe microglia response

 

A maximum intensity projection time-lapse movie showing Müller cells (in green) and Microglia (in red) in a 4 day post fertilisation zebrafish retina. Targeted ablation with a pulsed NIR laser allows the triggering of single cell death and continual imaging with 2-photon excitation.

This work is part of a research project conducted by Ryan MacDonald and Xana Almeida.