Collective noun for a group of electrophysiologists?

My husband and I have had an ongoing discussion for quite a while now about what the collective noun for a group of electrophysiologists should be. This discussion came up again a few nights ago, where he firmly believes that the collective noun should be a ‘superstition‘. I, on the other hand, think it should be a ‘paranoid‘, or maybe an ‘expletive‘.

I started my first career-related job recently. I’m now working as a manual (conventional) patch clamp electrophysiologist for a contract research organisation. It’s definitely a completely different world to academia and I’m having to pick up a lot of new concepts very quickly, but happily, I’m enjoying myself very much (despite having to write my thesis in the evenings and weekends).

Although it’s a whole new world for me, there’s definitely something oddly familiar about the environment and the people I work with.

I find myself smiling whenever I think back to my uni days, when I would sit at my patch clamp rig and fantasise about electrophysiology equipment that wasn’t held together with dog-eared autoclave tape or covered in aluminium foil that was torn in several places. I also used to dream about what it would be like to have a comfortable chair, a responsive computer and enough workspace to accommodate a keyboard, a mouse, an open lab book and a monitor (or 2 – if I was doing MEA experiments). On my first day at work it came as a little bit of a surprise to find that some things really don’t change.

I operate two patch clamp rigs now (not at the same time!) and although they are pretty much identical, they each have their own foibles. Whether it’s the optics of the microscope, or the amount of patience (and swearing) it requires to get the perfusion flow rate just right, or the extra stability a few centimetres of tape gives to the manipulation of the patch electrode, or the aluminium foil that was once wrapped around a cable but is now half hanging off and catches on the stage of the microscope. There are even uncomfortable chairs and very limited workspace! But thankfully I still get to work beside a window (yay daylight!) and at least the computers (and IT support) are much better now.

I sometimes used to think that maybe I’m a little bit too particular when it comes to conducting electrophysiological experiments. I have a certain way of doing things, often in a certain order, and I like my work environment to be organised and tidy. I also like to have particular (and in my opinion, essential) bits of equipment to hand in case I need it (ie., plastic syringes, forceps, super glue, copper wire, silver wire, tubing, foil, soldering iron, hot air gun, pipette glass, heat-shrink, blu tac, tape, lab roll, pipette tips etc etc). Having now found myself surrounded by highly skilled and very experienced electrophysiologists, I’ve discovered that maybe I’m not so weird after all. I may have a certain way of doing things, but I’m definitely not the only one. It also turns out that I’m not the only one who gets paranoid about flushing the perfusion tubing sufficiently, or the particular sequence of events that need to be followed in order to successfully patch a cell, or having too many people in the room when you’re trying to do an experiment.

There’s even a hint of slightly irrational superstition. One of my colleagues has said that their two best days, in terms of the success rate of patching a certain cell line, was when they were listening to music by a certain band. As we’re currently having our patience tested by this really troublesome cell line, it is now a firmly held belief that our chances of getting data will be massively improved by listening to this particular band. The only caveat being that my colleague was the only person in the lab at the time, so it may not work if the rest of us are there to watch/try it out. We all solemnly agreed that the conditions should be kept the same as last time, otherwise it will never work.

So this has provided my husband with more proof that a group of electrophysiologists should be called a ‘superstition‘, which is a good call, but I personally think the collective noun should still be a ‘paranoid‘ or an ‘expletive‘. Funnily enough though, I get really anxious when my husband puts new shoes on the table. I know it’s completely irrational, but I just can’t help myself. Maybe electrophysiology appeals to a certain kind of person.

attempted murder of crows

When the stress gets too much

It’s not easy being an electrophysiologist. Most of the time we’re under a lot of pressure and not just from our supervisors and managers, but from ourselves too. Sometimes we go for weeks, even months, without a single successful experiment. We know that it has worked for us before, but now that that all important deadline is coming up, electrophysiology refuses to play ball.

Very true

Very true

When asked “what’s wrong?”, most of the time the answer will be “I just don’t know”. This is not because electrophysiologists are lazy and have no idea what they’re doing. On the contrary, we’re very acutely aware that many, many different things have to be just right in order for an experiment to work – and sometimes even that is not enough.

Disaster can strike at any time

Disaster can strike at any time

Going through the list of things that have to be absolutely perfect can be time consuming, but any electrophysiologist worth their salt will have a vague idea of what the weak links are likely to be.

Some days are a bit 'meh'

Annoying

Some days the electrodes  are not absolutely perfect

Sometimes the problem can be really obvious.

Another experiment ruined and a day wasted

My experiment may not have worked, but I do enjoy a bit of DIY

I hate it when the perfusion system doesn't co-operate

Occasionally stress can be released in the form of exclamation marks and profanities.

Sometimes the tools you work with can let you down and spoil your day

And right after I put on some drugs too

Sometimes it's the computer program that can let you down

But when the stress gets too much, sometimes the best thing to do is walk away, have a cup of tea and calm down.

Next time you see an electrophysiologist alone in the corner with a cup of tea, approach with caution

Next time you see an electrophysiologist alone in the corner with a cup of tea, approach with caution

 

Be careful! Sometimes pent up stress can erupt when you least expect it:

A group of PhD students were in the breakout space one afternoon, taking a break from work. Everyone was happily chatting away and conversation turned to favourite Family Guy sketches. Someone quoted a particularly funny phrase and everyone laughed. One person in particular laughed so hard, he could barely breathe. Everyone starts laughing again and this time two more people laughed so much that they struggled to speak. The uncontrollable laughing continued, but not everyone was doing it anymore. Only three people were laughing, but their sounds of laughter started to sound more and more like sobs. There was a bit of an awkward silence and one by one people started to leave and return to the lab. The PhD students wiping their eyes and taking deep breaths looked round at each other and realised that they were electrophysiologists.

Like I said before: if we didn’t laugh, we’d cry and believe me, we’ve cried a lot.

You have been warned

 

For the love of brain (well, the hippocampus) II

So I’ve already described how much I love being able to look at live, functional neurons in real time. But there are other things I enjoy seeing everyday too, in particular my favourite part of the brain; the hippocampus.

The rodent hippocampus in stained, coronal sections

The rodent hippocampus in stained, sequential coronal sections

At the start of every patch clamp experiment, when I look down the microscope at a brain slice, I need to locate a specific structure before I can focus on finding individual neurons. This structure is called the hippocampus (so called because a cross section of it looks like a seahorse) and every brain has a pair of them (one in each hemisphere). The hippocampus is very special because it has a well-defined and distinctive structure, which makes it instantly recognisable to anyone who has studied neuroanatomy.

Freshly cut transverse brain slices. I store them in a plastic tea strainer, submerged in  carboxygentated aCSF. The arrow points to where the hippocampus is loacted

Freshly cut transverse brain slices. I store them in a plastic tea strainer, submerged in carboxygentated aCSF. The arrow points to where the hippocampus is located

 

Acute, transverse hippocampus as seen during my patch experiments - notice the patch electrode emerging from the CA1 region in the right-side image

Acute, transverse hippocampus as seen during my patch experiments – notice the patch electrode emerging from the top of the CA1 region in the image on the right

As I’ve said before, the brain is not a homogenous blob, but contains many intricate and beautiful structures. Many of these structures look pretty much the same across the majority of species, no matter what size or shape the brain may be, and the hippocampus is no exception.

Drawing by Camillo Golgi of a hippocampus stained using silver nitrate

Drawing by Camillo Golgi of a hippocampus stained using silver nitrate

Why do I love the hippocampus? Well, for one thing I love how ‘organised’ the anatomy is (I won’t go into detail here, but I do find the Wikipedia page extremely useful). The hippocampus is a network and I love how you can stimulate a particular pathway and get other neurons to fire in response (ie. stimulate the axons of the CA3 pyramidal neurons and the CA1 pyramidal neurons will produce a response). I also love the fact that it’s involved in lots of different aspects of learning and memory.

So, going back to the point about being able to stimulate pathways in the hippocampus, I also do a lot of experiments using multi-electrode arrays (MEAs). Being able to stimulate particular pathways and measuring evoked responses can tell you a lot about the synapse involved and the neurons on either side. For my experiments, I stimulate the Schaffer collaterals (axons of CA3 pyramidal neurons) and record the responses of the CA1 pyramidal neurons. Instead of observing signals from individual neurons, I look at the combined response of a whole population of neurons, which all fire in unison when stimulated by an extracellular electrode.

I take a photograph (before and after!) of every hippocampus I use for my MEA experiments. It allows me to select which electrodes to stimulate so that I can record evoked field potentials

I take a photograph of every hippocampus I use for my MEA experiments so that I can select which electrodes to stimulate in order to produce evoked responses. This is not a stained section – it’s the actual colour of a ‘living’ hippocampus!

I’d love to go into more detail about MEA experiments, but I’ll save it for another post 🙂

For the love of brain

What gets me out of bed in the morning and into the lab by 8.30am?

Answer: The love of neuroscience.

 

I freeze my aCSF in the shape of little hearts <3

I freeze my aCSF in the shape of little hearts

 

I absolutely love the brain. It’s such a complicated, yet incredibly delicate, structure and was the last major organ I learnt about during my 20+ years of full-time education. Of all the things that humans have discovered and achieved in the last few thousand years, the brain still holds a lot of secrets.

For every experiment I did during my PhD I had to cut very thin slices of fresh rat brain. The slices were 300 micrometres thin, which is just under a third of a millimetre (0.3 mm). They had to be this thin in order for light to shine through them so that I could see individual neurons for my patch clamp experiments.

 

This is what I see when I do my patch clamp experiments

This is what I see when I do my patch clamp experiments – note the patch clamp electrode approaching the middle cell from above. The cells are pyramidal neurons (because they are triangular in shape) from the hippocampus.

 

I really enjoy seeing actual neurons in situ. There’s nothing quite like it in my opinion. It got even better when I would successfully ‘patch’ a cell and see the wiggly lines on my computer screen form the distinctive shapes of action potentials, which are the electrical signals that neurons use to communicate with each other.

In the very beginning I found it slightly odd that the smooth spiky structure on the black and white screen was able to produce the electrical signals on the oscilloscope without looking as though it had changed in some way. In other experiments I’ve done that involved living tissue, there was always something that moved to show that it was still alive and functioning properly. But when it came to successfully patching a neuron in a fresh brain slice, I suppose I was half expecting the neuron to contract or flash or something. But no, it just stays there, unmoving until you’re finished with it (unless it’s very unhappy with having a hole in its membrane and decides to swell and die).

Neurons aren’t the only things I see in brain slices. A lot of the time I see red blood cells, usually in capillaries. I think it’s really cool and it also gives me a better sense of scale when comes to visualising how big neurons are (as it turns out, they’re bigger than red blood cells).

 

A capillary in a brain slice - pretty cool huh?

A capillary in a brain slice – pretty cool huh?

 

Being able to see living, functioning neurons in real time isn’t the only thing that gets me out of bed in the morning, but that’s another story 🙂

 

What not to say to a patch clamper

1. “It’s character building”

Often said when you’ve been banging your head against a brick wall for weeks, trying to get your experiment to work.

I agree, it is character building when you’re just starting out – I think it’s really important to instil the notion that patch clamp is incredibly difficult and frustrating. But when you’ve been doing it for at least a year with virtually no data to show for all those weeks and months of “character building”, then the person saying this is in danger of getting stabbed in the eyes with patch electrodes.

 

Death by patch electrodes

Death by patch electrodes

 

2. “I can see lots of cells”

The person saying this is either looking over your shoulder at a screen, or looking down a microscope at a cover-slip.

Yes, they think  they can see lots of cells. They assume that all those perfectly round, transparent blobs are cells that can be patched. But the truth is is that all those “cells” are in fact dead. Some are so dead, that all that’s left are ghosts of cells. When hunting around for that one reasonable-looking cell; a cell that can temporarily alleviate your misery, the last thing you need is someone with an untrained eye making completely pointless, unhelpful and incorrect observations.

 

3. “Are those miniature currents?”

Similar to the above, where someone (usually a PI) looks over your shoulder at a computer screen showing the latest trace of your cell’s electrical activity.

Once again, they think they can see miniature currents and yes, those currents are quite tiny. What they fail to notice, however, are the signs of a crappy cell, let alone the fact that the cell isn’t being perfused with any drugs (in order to isolate the miniature currents).

The same can be said for the relatively huge spontaneous currents. No, they are not inhibitory postsynaptic currents because I haven’t actually isolated them yet. I’m waiting for you to go away so that I can dash around the lab and prepare my solution before this cell dies.

 

4. “When will you have this dataset complete?”

How long is a piece of string? In an ideal world I can get 6 replicates (the minimum amount I need to run a statistical test) in a couple of weeks. The reality? A couple of months.

 

5. “Can you just do a quick ‘look-see’ experiment for Dr So-and-so?”

Seriously? You think that I can just do a ‘quick’ patch clamp experiment? For someone else? In a different part of the brain? With a completely different cell type and shape? On top of all the other experiments I have to do?

How about….. no.

Patch clamp