Nah, I don't think so. The work was pioneered by Konerth and Sakman and was done in immature rats at 1-2 month of age.

There are some papers in mice looking at somatosensory cortical cells, again in young animals.

In AD, pathology is usually heavy in entorhinal cortex and other cortices so that may be a good place to look for changes in the physiology.

well, I have recoded 2-3 years old mice but only in vitro see:

http://www.jneurosci.org/content/32/17/5891.full

no idea about in vivo recordings.

In vivo under anaesthesia, this is likely doable, provided you have either experience and/or years to work it out. Awake in vivo whole cell is always on the extreme end of what is possible, but it is still doable as long as you can get your pipette pointed at the target. Doing these types of measurements in the entorhinal (as suggested above) is somewhere between very and extremely difficult because of the location of this cortex, never mind if you want awake rather than under anaesthesia...much much worse if you want to go for free exploratory behavior in VR. Lateral EC might be a techically wiser choice for a target, although our in vivo understanding of spiking in the lateral versus medial is significantly less developed. The only in vivo recordings from the mEC in behaving animals were done before 10wks of age, and the goals of these papers were very specific (identifying neurons that produce grid patterns and possible clues to network mechanism), rather unlike what your aims appear to be. Be very sure what you want to measure will require in vivo whole cell...rather than in vitro whole cell or tetrode recordings in vivo instead. You could go in vivo whole cell after you have the phenomenon pinned down and just want the gold standard proof as the final nail.

What does "in vivo" mean in this question? Sometimes slice recordings are called "in vivo", but also "in vitro". This categorizing totally depends on the field of researchers. Especially, biochemists like to call slice recordings in vivo. John, please clarify to avoid further misunderstandings.

thanks for your answers, I know there is lots of people doing whole-cell patch-clamp in vivo in young or even adult mice, but older animals I can't find a thing. Sahar your in vitro experience might be helpful, considering the huge span you covered! How much harder did it get with age to get seals and/or break in? Did the quality of the prep deteriorate more quickly? Would be nice to know a bit more about that, also in vitro should give some idea. Cheers!

Some people routinely patch neurons in living adult rats (~200-300 g) if that's what you mean by in vivo. See for example: Constantinople & Bruno 2013, Oberlaender, Ramirez & Bruno 2012

You need to be prepared to accept a lower success rate. Instead of almost every pipette fetching you a cell with access in the 1-15 MOhm range, only a fraction of the pipettes will turn up a cell, with a larger range of initial access (e.g., 1-50 MOhm).

sorry I missed the last 2 answers before! By "in vivo" I mean the living animal, whereby I was thinking of an anesthetized mouse before trying anything more complicated like an awake prep.

Jonathan, I'm not sure what you mean by "provided you get your pipette pointed at the target". Are you saying it's doable as long as you don't go for a blind approach? I have thought of trying 2-photon guided patching in L2/3 parietal cx, but I no experience in that (only the blind approach).

I'm interested in confirming in vitro results from parietal cortex and hippocampus, by looking in vivo at things like spike shape, Vm, and PSCs if possible (given likely bad voltage clamp). We'd like to link these things with some EEG parietal cx data which is likely more general than just parietal cortex. So hippocampus (CA1 or DG) looks like our target, although LEC is an interesting idea (given how little is known, would be fun to look in AD but probably not the first place to look).

Any insights into just how much harder it might be? I find that in the anesthetized rat it's rather easy to get a whole-cell recording (in CA1), but the fact that there seem to be no publications out there is a bad sign. Is there any published patch-data from older rats in vivo?? There is just one paper with sharp electrodes from the Hyman lab (Stern et al., 2004), would be very happy to hear of anything else!

The oldest rats that I have obtained whole-cell patches in vivo were about 4 months. But I think old rats would be possible as well, but I imagine the stability and quality of these patches would be lower.

The only person I know of doing this is Dr. Claire Meehan in Denmark. She has been the modern pioneer for in vivo recordings from motoneurons in mice, and I believe she has recently been working in aged animals (I saw her present this work at a conference about a year ago, but I haven't seen her publication on the data come out yet). However, she is doing sharp recordings rather than whole-cell patch clamp recordings. You may be able to adapt her technique and she might have a few tricks to increase recording success rates in older animals. I imagine she might be a very good resource for any questions you might have. Her profile is here: https://www.researchgate.net/profile/Claire_Meehan2/

You may also want to look at the work of the Brecht group.

See for instance:

http://www.ncbi.nlm.nih.gov/pubmed/12971892

http://www.ncbi.nlm.nih.gov/pubmed/16908406

http://www.ncbi.nlm.nih.gov/pubmed/19247288

I've made whole-cell recordings from aged animals in a urethane anesthetized preparation. That being said, it is much more difficult and time consuming to get low-resistance seals with mature animals and may not be feasible. Your success will depend on what brain area you are interested in, because moving the pipette through tissue decreases your chances of getting a seal. Therefore, recordings will be more successful your target is superficial, such as layer 2/3 of the cortex.

This group uses adult mice with success: http://www.ncbi.nlm.nih.gov/pubmed/22561988

Also, as mentioned by Nicola, Michael Brecht pioneered this work and happens to be at your university! http://www.ncbi.nlm.nih.gov/pubmed/12136268

You want to look at David Tank's group out of Princeton.

Harvey, Collman, Dombeck and Tank (2009) Intracellular dynamics of hippocampal place cells during virtual navigation, Nature, Vol461, pp941.

Once you have that paper do a Science citation search on it (web of Science) or google scholar, and see who has cited that paper recently... odds are they will be doing the same thing. in the Nature paper aanimals were mice, and just barely mature. 8-12 weeks. But the technique will work in mature animals, just a lot lower yield, and greater difficulty in getting a good seal.

thanks for the great response! I am actually in the Brecht lab :), but we patch from rats and always relatively young (around p28) with very clear drops in success as the rats get older.. Thanks for the refs for 8-12 week old mice, but I'm thinking of really old mice (where AD symptoms pop up), anesthetized. It's clear that things get harder, but how much harder?? Timothy, it seems you're one of the few to have done this in really aged animals, but you're using hamsters??

Tim, Randy, Heather, great to hear you have this experience! Randy I had a look at the papers, great work and encouraging. Can someone give some (more) rough numbers on just how much harder things get with age? E.g. chance per penetration of getting a seal, succesful break-in? Rough average of penetrations needed to get a cell per animal?

Most importantly, what is overall succes rate per animal? I would assume the prep rundown is worse in older animals (??), so if you combine that with low succesrate per pipette there's a risk of just getting nothing.. Other questions of interest are typical access resistance range and recording length. Maybe I'm asking a bit much, but any specifics would be great. Cheers!

Funny. If you are in the Brecht lab...why are you asking us? For sure, as one of the few labs having ever achieved whole cell in vivo awake behaving...your lab must have the know-how somewhere in there.

As far as my previous comment regarding targeting your recordings, again...being in that lab, you should have already been well aware of the challenges of getting a pipette pointed at the MEC. It is not like finding the barrels with a skull window, for example. The MEC is in a horrible location, as those who have managed it will tell you.

Success rates in whole cell (in vitro or in vivo) are all dependent on you when you choose to do something this obviously difficult.

Once again, I would suggest trying to make this your closing figure data, rather than your meat and potatoes experimental plan.

Hi John, sorry for my late answer.

my tricks to record old cell: a smaller pipette is the better, I usually keep the resistance between 6-7 for older cells. as these cells are more fragile a smaller pipette helps not to ruin them when you are breaking through. I used to prepare my slices with 350 micro M thickness. patch clamping is easier in thinner slice as the animal gets older. May be because it is easier to recognize the cells. therefore I did 250 instead of 350 for my aging project.

the old slices and cells look as good as the young ones and the stay a day long healthy. However old cells are more sensitive as soon as you start poking them!

If you succeed to make a seal and break through then the cell stays with you as long as a young cell.

I hope I could help, let me know if you need more information.

Sahar

Talk to Jonathan Ting (here on RG). He has protocols for whole-cell in older animals.

Thanks to Daniel for the plug...

Hi John,

I have found that there are several relatively simple modifications that can be made for the acute slice protocols that allow very routine successful patch recordings across many brain regions in adult and aging mice (see ). However, it is hard to imagine how to apply this to in vivo recordings and hard to imagine that the cells I am patching could be any more healthy (they are almost certainly less so) than cells in older anesthetized rat in vivo (in the intact brain). I can say that it is possible to make slices from old animals that are about as easy to work with as the typical slices from juvenile animals when applying the methods I have referred to. If the levels of glutathione are restored to high levels the slices from old animals can also remain stable and healthy for extended time frames for the experimental session. So hopefully this hints at the strong possibility of related or as yet unknown methodological improvements that could also work for improving the success of in vivo recording in old animals.

It sounds to me as though the deterioration in cell viability over time with older animals in the anesthetized prep may have more to do with the effects of the anesthesia itself than the true stability of the cells in the intact brain. Perhaps a closer look at adjustments to anesthesia that may be more gentle for the less resilient aging animals would be a good place to start. This is analogous to slice physiologists figuring out that brain tissue from older animals was less tolerant of large temperature and osmolarity fluctuations during the slicing procedure (e.g. work by Moyer and Brown). With some minor adjustments the success rate went up significantly.

I did sharp micro pipette recordings in hippocampal slices from 20-26-month-old rats and mice. Despite cells from aged animals are more fragile than younger ones, recordings are successful. I hope sharp pipette recordings work also in vivo.

I am doing two-photon targeted paired whole-cell clamp (TPTP) in adult mice. Success ratio is not significantly different with juvenile mice, in TPTP. But in my slice experiments that was significantly different. I think one of the important issues is cell visualization, using fluorescence dye and proteins are helpful for doing TPTP in aged mice. Another important issue is increasing of pipette resistance in patching cells in deeper areas of  brain of aged mice, in vivo, one solution is using more positive pressure.