I have written about the use of fibre photometry to record Ca2+ activity in vivo, and today I’ll be exploring a more advanced (and far more complex) version of that. Namely, the use of a head-mounted miniscope to record videos of individual neurones.
I first learned about head-mounted miniscopes at the same time as photometry – in 2015 when Chen and Betley showed how AgRP neurones really work1,2. Nobody could read the Betley paper with their beautiful head-mounted miniscope data, and not be excited by that data and want to do it for themselves.
But, one must also recognise that it is clearly an exceptionally complex technique, and that you should only use it when you absolutely need to, ie. don’t do the super-difficult version when you can get just as good an answer with fibre photometry. And having said that, I don’t know if miniscopes were necessary for Betley’s paper – Chen found many similar results without them.
Anyway, my point here is to reiterate what I always say, which is to make your experiments as simple as possible, to give you the strongest and cleanest answer. So in that vein, I will investigate a paper that used miniscopes to find a response that wouldn’t have been possible using photometry, a 2018 paper by Chen et al.3
This paper combines head-mounted miniscope recordings of Galanin-expressing neurones (Gal-cre) of the dorsomedial hypothalamus (DMH) and telemetry-based EEG recordings of brain activity (Figure 1A). They combine the data to allow them to correlate the EEG activity showing different phases of sleep/wake and GCaMP signal from individual neurones (Figure 1C). What’s really interesting is that they show two distinct subpopulations of Galanin neurones, with opposite behaviour during REM and non-REM sleep.
So they performed a series of exhaustive tracing studies (which I won’t go into here), that showed strong and mutually exclusive projections from the DMH galanin neurones to the preoptic area (POA) and the raphe pallidus (RPa). To show these correlated with the REM and non-REM sleep patterns, they redid their miniscope experiments on the DMH, but this time they used a retro-transported AAV-GCaMP to label specifically the differently projecting subpopulations (Figure 2A/E). This elegant experiment showed that the POA-projecting subpopulation was active during non-REM sleep (Figure 2C/D), but the RPa-projecting population was active during REM sleep (Figure 2 G/H).
The authors then go on to perform another exhaustive series of experiments, this time using optogenetics to show that the different DMH projection sites don’t just correlate to REM or non-REM sleep, they can also drive changes between those sleep states.
Lastly, I’m just going to briefly go into my interest in doing these experiments myself. A year or so ago, I enquired with Inscopix (who make the benchmark miniscopes, and I think were spun out from the lab that originally developed them) about purchasing one from them4. The quote came to £60k, which was far too much for us, so I forgot about them for a while to focus on other things.
And then recently, while exploring options related to developing fibre photometry, I came across the open source head-mounted miniscope project from UCLA5. I had seen this before but the sheer complexity put me off. Essentially, they have developed their own miniscope, and have made the designs freely available online. The problem is that this is such a complex technique, I wouldn’t be happy having to build the microscope myself as well as learning and optimising the system; I could just see it being a massive waste of time to get it working well.
Anyway, when I revisited the UCLA miniscope site recently, I found that they have not only released a new lightweight and more advanced version of their miniscope, they also have started selling them fully assembled on the open ephys website6. And their price? £1,940 (including the acquisition box). So, needless to say, I will be requesting from my supervisor that we buy one. Or five. The price is reasonable enough that I think the only reason he’ll say no is if he considers it a waste of my time. Or more to the point, that playing around with one of these will distract me from my -real- work.
There is major challenge with getting a miniscope from anyone that isn’t Inscopix, and that comes down to the GRIN lenses that you need to do the imaging in the brain (for any that don’t know, the GRIN lens is like a fibre optic that has a precise structure that means you keep the image in focus). Anyway, it turns out that the only company in the world that makes GRIN lenses longer than about 4 mm of a type that you can use for in vivo imaging is called GrinTech, and they have an exclusivity deal with Inscopix. Which means that they won’t sell them to you, you need to go to Inscopix, which means spending £60k.
So, for any “real” neuroscientists that work on structures such as the hippocampus or cortex near the brain surface, you should be fine to get the cheap miniscope and get shorter GRIN lenses from places such as Edmund optics. I, on the other hand, and anyone else who works on more interesting and deeper brain regions, will have to keep searching.
1. Chen et al., Cell 160, 829-841 (2015) Sensory detection of food rapidly modulates arcuate feeding circuits.
2. Betley et al., Nature 521, 180-185 (2015) Neurons for hunger and thirst transmit a negative-valence teaching signal.
3. Chen et al., Neuron 97, 1168-1176 (2018) A hypothalamic switch for REM and non-REM sleep.