Ideas for measuring cell thickness and Ph level ...
Beau Schwabe
Posts: 6,568
A friend of mine is heading up a research study this summer involving the promotion of 'cell' growth.
Tonight he was asking me if I had any clever ways that I could think of to measure/monitor the thickness (growth density) of the cells. Also ways to measure and monitor micro Ph levels during the normal growth cycle of the cells.
Just thought I would bounce the idea off of the forum and see what turned up.
Note: This is not 'stem-cell' research ... this is strictly using existing healthy cells to promote new growth.
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
Post Edited (Beau Schwabe (Parallax)) : 5/29/2010 2:53:03 AM GMT
Tonight he was asking me if I had any clever ways that I could think of to measure/monitor the thickness (growth density) of the cells. Also ways to measure and monitor micro Ph levels during the normal growth cycle of the cells.
Just thought I would bounce the idea off of the forum and see what turned up.
Note: This is not 'stem-cell' research ... this is strictly using existing healthy cells to promote new growth.
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
Post Edited (Beau Schwabe (Parallax)) : 5/29/2010 2:53:03 AM GMT
Comments
If you've got cells that phagocytize tiny particles, I think there might be nanospheres that change color with respect to pH.
This isn't the article I remember reading, but I suppose it's the same concept:
www.rsc.org/publishing/journals/OB/article.asp?doi=b900260j
If he's not looking for anything too advanced, I'm in the middle of making an optical densitometer from the TSL230. I have no idea yet how well it will work, but it's probably a reasonable device to tinker with. The density of cell growth should affect the passage of light through the sample in a meaningful way, I hope.
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Watching the world pass me by, one photon at a time.
Post Edited (ElectricAye) : 5/29/2010 4:13:04 AM GMT
it was On or Off .
but with a analouge photo transistor or what not you could find a way to shine a precice amount of light thrugh the cells and see how much is lost with transmission .
and cause your not counting cells just the thickness a larger tube might work ..
it would would work BUT it would need a Very presice mesurement ..
If the buffer fluid with no cells = lets say 37 and the FULL healthy cells with the fluid is like 165 then just - the Fluid mesurement in the MCU to get the actule cell reading ..
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If its In a petri dish then reflective Might work
I am not a Bio major [noparse]:)[/noparse] but I remember a bit . May I ask what kind of cells .
Eg a amouba do to its non uniform shape would not lend its selfe to passive mesurememts Like this ..
Peter KG6LSE
EDIT ElectricAye HAHAHAH we had close ideas at the same time ..
Great minds Do think the same ..
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"Carpe Ducktum" "seize the tape!!"
peterthethinker.com/tesla/Venom/Venom.html
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LOL
Post Edited (Peter KG6LSE) : 5/29/2010 4:18:36 AM GMT
The cells are human cells located within the trachea. That's about all I know right now.
I might add, that the cells must remain intact and whatever method is applied cannot be destructive to the cells.
The most recent application area that he has worked with involves the development of regional angiogenic imaging bio-markers for cancer.
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
This might seem like a stupid question, but are you talking about monitoring cells that are located within the trachea still inside a living human or are you talking about monitoring cells that are from the trachea but are grown in vitro, inside something like a cell culture dish outside of a human?
"Cell organelles are the tiny functional units that exist in cells – examples include the nucleus, the mitochondria, the chloroplasts, the ribosomes and the golgi body and endoplasmic reticulum (rough and smooth). A typical animal cell has a diameter of about 50 micrometers – a micrometer being one thousandth of a millimeter. Cell organelles all fit inside the cell and are therefore all much smaller than this. The measurements we use to say how big they are include the micrometer but we also tend to use the nanometer, which is one thousandth of a micrometer. This allows us to more easily describe the sizes of the smaller organelles. For example, a cell nucleus can be about 10 micrometers in diameter but a ribosome is about 25 nanometres across. This is easier than saying 0.0025 micrometers. A cell and the major, larger organelles can be seen with a light microscope but the smaller ones need to be viewed with an electron microscope. "
http://www.blurtit.com/q538934.html
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http://www.bdbiosciences.com/nvCategory.jsp?action=SELECT&form=formTree_catBean&item=378205
You might need a cell strainer:
http://www.bdbiosciences.com/nvCategory.jsp?action=SELECT&form=formTree_catBean&item=619502
I'm not trying to sell you anything but I know people who sell these items.
"This might seem like a stupid question...are you talking about monitoring cells that are from the trachea but are grown in vitro, inside something like a cell culture dish outside of a human?"
Not a stupid question at all. They are grown in vitro with the intention of grafting them as a functional replacement similar to skin grafting for burn victims. This is why any testing or monitoring of the cell growth progress can not be damaging or irreversible to the cells themselves.
Like I said though I'm not doing the study, my friend just asked me if I had any ideas that I thought might work. This Dr has specialized over the years with pioneering various bio medical imaging techniques, i.e with 3D CT scan image processing, so I think he is very capable and has access to lots of high dollar equipment, but for this particular study he hasn't quite worked out the method he is really comfortable with that won't harm or contaminate the sample in any way.
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
Sounds like a cool project. Do you know what he's growing them in/on? Is this a monolayer of cells grown in tissue cell culture flasks or is he growing them on some kind of bio-absorbable substrate? Because mammalian cells grown in vitro tend to grow in a single layer, I'm thinking a monolayer of cells grown in a flask would be hard to harvest in a sheet form that can be grafted. In any case, I'm told that counting cells in flasks is either done by hand using a microscope or it's a megadollar robotic project, but I' wondering if a monolayer of cells might interact with monochromatic light to create some kind of interference pattern that could be evaluated somehow. Perhaps a holographic "mask" of some sort could be developed and laser light could be bounced around between/through the mask and the cell layer. When the cell layer reflections/refractions match some sort of "recognizable" density/pattern, then an interference signature could ring a bell. But please don't be fooled: I have no idea what I'm talking about here, but I thought I'd throw that out there for people to think about.
If the cells are being grown in some kind of substrate, then an optical approach might be more problematic as the substrate itself might muck things up. As always, the devil will be in the endless details.
Engineers credo: more info, please.
http://www.lrsm.upenn.edu/
http://webdev.chem.upenn.edu/chem/facilities/xray.php
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
Of course, I'm not sure this would be a wise way to do it for a product going into humans (unless the mild radiation is deemed acceptable vs. having no trachea), but it might work well enough for helping out with the initial research.
kwinn might chime in here. I bet he could comment on an approach of this sort.
Anyway, just a thought.
Thanks! ... this area is totally out of my comfort zone, although my original studies were Pre-Med Engineering. With Dee's permission, I thought I would toss the concept out here in the forum to see where it would go.
Small world story... I met Dr. Dee Wu about 3 years ago at a school function (our daughters were both in the same class) We randomly sat down at a lunch table and started talking casually... When he found out I worked for Parallax, he nearly jumped out of his chair. (<-- really it almost scared me a little) He teaches on the side and has used Parallax products in class and was very familiar with us. Dee is a smart guy with a very impressive bio, and he will find a way to get the job done, but I was honored that he asked me for any ideas involving this project.
So far I have discussed very sensitive differential capacitive techniques where the cells themselves become the dielectric. Another method which falls along a similar category with measuring the Ph level is to measure the resistance of the cells with very low voltage and current, again in a differential approach to increase the signal to noise ratio and filter out any noise.
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Beau Schwabe
IC Layout Engineer
Parallax, Inc.
Post Edited (Beau Schwabe (Parallax)) : 6/4/2010 5:21:08 AM GMT
Using low level radioactive tracer molecules is not unusual in cell culture labs. And using a spectrometer/detector is akin to giving your man-made trachea a CT scan. You can make a DIY spectrometer using a photomultiplier tube optically coupled to a material (crystal or plastic) "tuned" to whatever sort of tracer you want to feed the cells. I've built a crude spectrometer from junk off ebay and used the Propeller to handle all the controls, scanning thresholds, etc. Different detector scintillator materials and maybe shielding are used for different kinds of tracers. As for developing it "at home", the biggest drawback would be in testing anything radioactive. Yes, you can legally get low level radioactive sources to test things without any special certification, so you could get some rudimentary bench top testing done in your shop without having to wear lead underwear, but for actually feeding the cells, etc, you would need a certified lab, license, proper disposal, etc. But I'm guessing that level of testing would have to be done at the research lab anyway, and I'm sure they would have the proper certification for handling the hard core radioactive stuff.
Of course, if the guy has funding, you can probably find nifty off-the-shelf radiation detectors, so most of your hardware problems would be solved by that.
As for passing electrical current to measure cell resistance.... I'm guessing these kinds of cells are similar to muscle cells(?) so be sure to do your homework on what sort of effect small currents might have on the cells. As I understand it, many types of mammalian cells simply won't continue living unless they are in touch with their neighbors, so they are very sensitive to something (chemical? electrical?) that tells them they are part of the larger group (and not just floating around in the blood stream, for example, where they aren't wanted). If you are passing voltages through their hand-holding survival routine, you might make them feel that they are all alone in the world and cause them to die.