Patch Clamp - How do you study Cells?
What have I been doing this week that kept me so incredibly busy?
Typical university stuff I suppose. Another lab practical, although this one was a bit more intense than the others (I didn't believe that's possible!).
The goal of this lab practical was to teach us the patch clamp technique, which is used to observe ionic current in living cells.
I can almost hear some of you thinking "what the fuck is that supposed to mean? And why would anyone want to do that? Biologists are damn weird". And yes, I agree, biologists are weird people. But the patch clamp technique has an actual use.
Cells aren't closed systems. They stand in constant contact with their surrounding cells. Several of these communication methods work with channels inside the cell membrane, which let ions pass through. That causes all sorts of things, explaining all of them in detail would take ages and probably bore most of you. Instead, I'll just say that we really want to know what's going on in there!
To use the technique, I first needed some living cells, preferably neurons. Acquiring those was not very pleasant, as they came from the olfactory bulb of the mouse you see above. Of course, I needed to kill the mouse. Awful way to start the day.@suesa
After spending some time taking everything apart and doing some cleanup, I was able to isolate the olfactory bulb.
With the razor blade, you can see at the top of the picture, I cut several thin slices. It was important that everything was really, really cold all the time so the cells wouldn't die off.
Here's a close-up of one of the slices, photographed through the lense of a microscope:
It's not just the olfactory bulb, I also got a good portion of the brain in there.
The slice was then placed under a special microscope which has a constant influx of a "nutritional solution" to keep the cells alive. There are also two electrodes and a tiny, tiny glass pipette.
The pipette is actually self-made! We have thin glass pipes that are put into machine with a heating spiral. The glass is then heated to about 60°C. At the bottom of the pipe is a weight. When the glass reaches a certain temperature, the weight pulls the lower part to the bottom which stretches the glass until it seperates. What's left are two shorter pipes with very thin tips.
Fancy, isn't it. It's actually funny how much "self-made" and "really fucking expensive" clashes in the lab all the time. In one room, we have a microscope that cost several thousand euros. And in the other room, I've been working with this setup:
I'm sure several parts of this are older than me. Especially the VHS player. And look at this:
This syringe (the blurry thing at the bottom) is attached to a tube that's connected to the glass pipette. Its purpose is to supply a certain pressure so it's easier to manouver the pipette through the tissue.
So you put some liquid into the pipette (should be a mix that's ok for the cell because it's going to be pumped into the cell), attach the pipette to the microscope, lower it into the tissue and closely watch the TV as everything is projected onto it. Then you press the pipette against a single cell, suck it in slightly and open it by creating a small vacuum. If you didn't fail (oh how many times I failed and destroyed the cell), you have now control over the voltage of the cell's membrane and can measure fun stuff.
Block channels! Shock the cell! Add poison!
All fun until you do it for 4 hours straight. 3 days in a row.
The scientist's life I guess!
Pictures were all taken by me
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Now I understand where your SF stories come from! :)
Being into science helps :P
Omg you are doing patch clamp
I was told that it is one of the most difficult but tedious technique, and people have to be trained for months before they can really move to the real experiment with reliable results.
I can tell you one thing: After a week of doing it, I still mess up most of the time before I even TOUCH the cell.
I guess you will be in great demand after equipped with this technique XDD
This is probably the only way to characterize ion transport via channel (?)
Btw after hearing “patch clam” stuff for years finally have a chance to see some nice pictures of it ~
Very interesting! I have also worked with optical tweezers to study cells (but as a physicist so dont ask me what it was about), but it it pretty difficult because the cells can easily die if you overheat them when using the laser.
My girlfriends is a microbiologist and her friend is also. That friend will use this technique to study some interaction between bacteria at different lengths from each other. I forgot the interaction unfortunately.
Cool! So I’m one of those people who wants to know “why”. I know you have a short answer above but could you expand it a little. Why are we interested in what’s going on in there? Will this help with some type of cancer research etc? What are you recording when you make observations of the cell activity. Thanks again, this was cool.
Knowing how cells communicate and how to manipulate that has several possible applications. Neurodegenerative diseases could be healed or at least treated. Pain therapy is another thing that comes to mind.
Cancer not necessarily in this case, as it's mostly a technique to study neurons.
Oh and about the recording: we record things like "do sodium ions leave or enter the cell? How much? If not, why?"
Sehr sehr coole Laboreinblicke!!
The scientist's life ain't really as boring as the Big Bang Theory led many to believe :)
It is always amazing to take a peep at behind the scene and get our
eyeshands "dirty" and see how the results that are neatly tabulated in science journals are gotten.I wanted to know something, does this test provide a different result from what is already known?
No, we just reproduced already known things. It was a practical intended to teach us how to use this technique. You won't believe how many glass pipettes I broke during this week..
Ok. I can understand, having broken quite some few things in our own lab :)
Great post! I only learned about patch-clamp in my textbooks but I'd love to see that irl. What is your field of study? (I know, biology, ofc, I mean more precisely :D )
Human- and Molecular Biology!
Yey! We're on the same boat! :D ... what about your bachelor or master degree thesis? Or already a phd?
I hope to finish my bachelor thesis in June :)
Excellent post to introduce people to patch clamp, especially because it is a bit forgotten technique in biology. It reminds me of time when my colleague and I tried to do a patch clamp experiment with cancer cell lines, we had a lot of fun!
Definitely an upgrade from labs done at high school. This really is interesting! Studying nature in a lab is always a thrilling task. Good to see you're still going :)
I read through the whole thing.
I was too biological for me.
Which living thing did you use as a specimen?
A mouse, as mentioned in the text.
Takes a lot mental strength to dissect a living organisms.
Right?
Being a vegetarian, I know how hard it is.😊
The killing is the hardest part. I don't like killing animals. As soon as it's definitely dead, it's easier. I know I can't hurt it anymore so there is less inner conflict.