Acute brain slice electrophysiology

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It’s been a busy, but at the same time really exciting, few months for this acute brain slice electrophysiologist. Presenting a poster, which was featured in the Imagining Science blog, at a huge international conference (Society for Neuroscience 2013) last November reminded me that there’s life beyond the four walls of the E-Phys. lab. It was great in the sense that I felt like people (who were not my supervisors or colleagues) really wanted to know about my research – a feeling I’ve never had before.

Not long after the end of the festive holiday, just as the fuzzy warm glow from SfN had started to wear off, I gave a 10 minute oral presentation at the Pharmacy Postgraduate Research Showcase 2014. Even though I presented the same piece of research that was on my SfN poster, it was still quite a challenge to fit it all into a coherent 10 minute talk. However, I was pleasantly surprised by a number of things that day. Firstly, half way through the presentation I started to enjoy myself – it’s not every day that you get to talk about your research to a captive audience of 50+ scientists. Secondly, at the end the presentation I was asked a lot of very good questions, which allowed me to expand on some of the points I had to briefly gloss over due to time constraints. Finally, I won the prize for the best 2nd year PhD student talk!

With this recent boost in confidence I’ve now decided to take part in Reading Science Week’s Science Slam where I hope to introduce to people the very interesting findings from my patch clamp experiments.

Acute brain slice

Image by Thomas Hill

 

Scanning Electron Microscopy (SEM) of 3D Neural Cultures

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The University of Reading EM Lab (formerly know as CfAM) has been assisting in our studies of 3D human neural cell cultures derived from a type of neural stem cells (normal human neural progenitors – NHNP). Here at Reading I’ve  been investigating the electrophysiological properties of cultures of neurons and glial cells differentiated from NHNP, grown in a 3D substrate called Alvetex. In order to see how cells are expanding and morphologically orientate themselves in Alvetex, I’ve been using immunofluorescence and SEM. Not only have the EM Lab done the hard graft of the SEM work, one of our images went up in their image of the month hall of fame for 2013. All part of the art of electrophysiology!

SEM (left) and immunofluorescent staining (right) NHNP cells grown in 3D scaffold SEM imaging: Matthew Spink @ EMLab, Immunostaining: Immy Smith @ UoR ERG. Cells – Lonza NHNP, Scaffold – Alvetex by Reinnervate, Antibodies – B-tubulin III (Millipore) in green, GFAP (Dako) in red, hoechst 33342 counterstain and secondary antibodies from Life Technologies)

 

This scanning electron micrograph shows a normal human neural progenitor cell (supplied by Lonza) which has been false coloured in blue. A normal human neural progenitor cell is a type of stem cell which forms neurons and glia in the brain during development. This cell has not yet commenced differentiating into neural lineages, and is still in the stem cell stage. The cell is grown in a 3D culture scaffold called Alvetex. This work is being undertaken by Dr Immy Smith who is part of the Electrophysiology Research Group in the School of Pharmacy. The work forms part of a project investigating functional differences in these cells grown in 2D vs. 3D to produce models for neurotoxicity and pharmacological studies. Sample preparation and microscopy assistance was provided by Matthew Spink.

DIY Electromyogram

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Just back from Society for Neuroscience (http://www.sfn.org/annual-meeting/neuroscience-2013) which was a five day fest of all things neuro and feel very much revitalised about our work. Aside from the forthcoming posts about more research-relevant things arising from SfN, I was delighted to see that Backyard Brains (https://backyardbrains.com/) had a stand at the conference as I bought one of their very early ‘build your own ‘ Spikerbox’ kits (https://backyardbrains.com/products/diyspikerbox) a year or so ago; the kids and I had great fun doing some earthworm and other invertebrate experiments and we built a cheap and cheerful Faraday cage from an XBox360 case!

So taken by their latest offering, I immediately snapped up their EMG Spikerbox kit (https://backyardbrains.com/products/emgspikerboxkit), complete with conductive gel, cables, electrodes etc. and have been looking forward to playing with this on my return. True to my word, I unpacked and – in a highly jet lagged state – worked my way through the muscle action potential experiment just to get familiar with the kit (https://backyardbrains.com/experiments/muscleAP). It’s another really excellent piece of kit and am now struck by the idea that it could make a fantastic MPharm project that develops new experiments that could be taken into schools as part of outreach events. I wonder if I can persuade any of my project students to rise to the challenge?

Anyway, lots more to do with the kit this weekend as I finally got Audacity configured to acquire the spike data and so now need to start spike detecting and sorting (see http://wiki.backyardbrains.com/Data_Analysis for details of how to get data onto PC/<insert name of device>).