Acute brain slice electrophysiology

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

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.