An Introduction to Microinjection

Microinjection is one of the key techniques I will be using during my summer project. It is an incredibly versatile and widely used technique, with applications from ranging from IVF to producing transgenic organisms. Its application during my project will be to introduce the gold nanoparticles (GNPs) into the cells for photothermal imaging.

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TEM of VEGF SmartFlares after uptake

Throughout all of the fluorescent microscopy experiments, we’ve been seeing SmartFlare fluorescence in punctate structures. Using these techniques, it’s very difficult to say with certainty that the SmartFlares are within vesicles or (for example) aggregated/agglomerated in the cytosol. As this is pertinent to the current investigation, we took cells loaded with SmartFlares to the electron microscope which has the resolution to discern membrane structures as well as the gold particles.

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Imaging inside cells with nanoparticles – a BBSRC-sponsored summer project

Hello! My name is Charlotte Dawson, I’ve just completed my second year studying Biochemistry at the University of Liverpool. Over summer I’m undertaking a 10 week research experience placement funded by the BBSRC, co-supervised by Dr Raphael Levy and Dr Violaine Sée.

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Direct Microinjection of SmartFlares

We’ve so far, not seen any response of the VEGF SmartFlares to treatment of our cells with DMOG. Combined with the punctate distribution of the signal, we have no evidence that the SmartFlares make it to the cytosol in 18 hours. As we’re not sure if the problem lies in the SmartFlares or the delivery, I microinjected the SmartFlares directly into the cells thus bypassing the need for endosomal uptake.

Microinjection
Image Courtesy of the Liverpool Centre for Cell Imaging

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