[removed]

[deleted]

[removed]

[removed]

It will not sterilize your sample since the dose rate is far too low. It will kill a few bacteria but most will survive and produces a new generation. It might lower the overall population a little for a short time but considering the adaptibility of bacteria to their environment it wouldn't take long for them to adapt.

What if the bacteria/fungus in the starting sample were unable to propagate in the liquid? Either no food or maybe even frozen for the duration of the "treatment"?

For all intents and purposes, no. This is roughly the dose rate of simply being on a plane at cruising altitude. There are plenty of bacteria that can survive living in radiation environments substantially worse than that indefinitely.

For context the standard dose for sterilizing medical devices is ~25 kGray or 2.5 million rad which is very roughly (neglecting for this back of envelope calc the difference between absorbed dose and effective dose) 2.5x10^10 microsieverts. So I mean if you wanted to wait a couple hundred thousand years...

[removed]

[removed]

[removed]

[removed]

>This is roughly the dose rate of simply being on a plane at cruising altitude.

My initial research suggested this rate was 5 times that of cruising at altitude, but your point is taken. Thank you.

I think that freezing should allow one to achieve sterilization with a low emission source by basically tilting the replication rate to kill rate in a severe way.

Low emission rate sources need more time to achieve sterilization.

Still though, depleted uranium generally provides alpha and beta emissions with a bit of gamma. The alpha won't make it through the walls of the ampule. I think that much of the beta also won't make it through. You'll be dependent on the low gamma emissions to slowly achieve your sterilization.

In terms of magnitude, 5x is about the same given the range of emmisivity that is observed with radioactive sources.

When one considers sterilization, one is looking for magnitudes of "kill". For instance, cooking chicken to FDA mandated temps results in log -6 reduction. One in 1 million salmonella survive the cooking process. I think that dental steam sterilization practices are going for log -9 reduction.

A multiple of 5 is not a considerable effect in the consideration of sterilization.

I think it's indeed a bit higher than the background radiation in that altitude, but something like an electron beam irradiator can output as much as 11000 Gray per SECOND, the dose required for sterilization is pretty high. Also, in some places like Ramsar (Iran) and Guarapari (Brazil) the background radiation can be as high as 40uSv/h but that's pretty rare

It's important to remember that cells are only really vulnerable to radiation damage when they are actively dividing because it's only in this stage that the DNA fragments are exposed and aren't attached to the 'scaffolding' used to securely store DNA in the nucleus. And when the DNA is stored it is readily repaired by the cell's repair mechanisms. (The DNA can still be repaired during cell division but the repair process is much easier to disrupt.) So you would have to concentrate alot of radiation energy directly into the stored 'DNA bundle', to cause enough damage to the DNA and the structure used to support it in order to overwhelm a cell's repair mechanisms. The only way to do this at this dose rate is to have the stored DNA hit by an alpha particle but the range of alpha particles from radioactive decay is so low that none would make it to your sample bacteria.

So if the bacteria are unable to move due to a lack of nutrients, and so not actively dividing, but are still at a temperature typical of their environment they would be incredibly resilient to radiation damage and you are unlikely to sterilize the sample.

Now if they were frozen it would be possible for the bundled DNA to accumulate enough damage over time to kill the bacteria once they were thawed but again not in any reasonable time frame with this does rate.

Fun fact: people who are cryogenically frozen will accumulate enough DNA damage from high energy cosmic radiation that they will effectively receive a lethal dose of radiation before being thawed if they are frozen long enough. Probably not the best financial investment over the long haul. #justsayin

edit: removed 'any reasonable time frame' from the second paragraph. / added 'at this dose rate' in the first paragraph.

>I think that freezing should allow one to achieve sterilization with a low emission source by basically tilting the replication rate to kill rate in a severe way.

>Low emission rate sources need more time to achieve sterilization.

>Still though, depleted uranium generally provides alpha and beta emissions with a bit of gamma. The alpha won't make it through the walls of the ampule. I think that much of the beta also won't make it through. You'll be dependent on the low gamma emissions to slowly achieve your sterilization.

The 15 microsieverts per hour measurement should already take that into account. The uranium sample itself is in glass so all of the alpha and most of the beta should be contained.

[removed]

First, I love this post and this is why I follow the sub.

Second, are you performing any experiments that have been, potentially, been contaminated?

Sorry, I misunderstood your use of the term ampule. I figured that your sample of bacteria was in the ampule rather than your uranium was encapsulated in glass.

In any case your work will be done with a weak gamma emitter which will probably work damnably slowly.

Essentially I made a piece of jewelry containing a concentrated solution of fluorescent protein. It began as a sterile solution and a sanitized glass ampule, but a mishap during the final step sealing it up (with me breathing over top) may have unfortunately introduced some contamination.

I didn't include any toxic preservatives like sodium azide for safety reasons in case it ever breaks.

It may be that a thorough irradiation sterilization will end up denaturing your protein anyways.

Maybe one could accept that a beautiful thing could be beautiful also because it is temporary.

At what temp will your protein denature? If the protein denatures at a high enough temp, you could hold it at a lower temp to make life really hard for bacteria and wait it out for a few days to kill it down while not denaturing your protein.

Basically pasteurize the thing at a temp that is below the temp that will damage the protein you care about.

[removed]

[removed]

No worries about the denaturing, fluorescent proteins are notoriously stable structures. They are even resistant to Proteinase K... which is ridiculous.

[removed]

>The radioactivity is due to the local geology. Underground water dissolves radium in uraniferous igneous rock and carries it to the surface through at least nine known hot springs.[15] These are used as spas by locals and tourists.

Uh. That feels... unsafe.

[https://en.wikipedia.org/wiki/Ramsar,_Iran] (https://en.wikipedia.org/wiki/Ramsar,_Iran)

[removed]

With enough radiation though you would create a large number of ROS which can chemically stress the cells and kill them in all sorts of ways.

> but most will survive and produces a new generation

Might be fun to feed those bacteria on purpose, and see how well they mutate and evolute towards radiation-hardiness!

[removed]

[removed]

Risks of low doses and low dose rates, such as from elevated natural background radiation exposures, appear not to exist or be lower than such risks that one assumes by applying the LNT model in the evaluation of epidemiological data. This and the unequivocal evidence of experimental findings of adaptive protection speak against the LNT hypothesis, which should be replaced by a model that takes into consideration that low doses can induce alterations in the physiologically individual balance between cancer causation and cancer prevention.

Source: Cancer Mortality Among People Living in Areas With Various Levels of Natural Background Radiation

Why not just heat sterilize it? If you do that in a pressure vessel the pressure inside your jewelry and surrounding it will be the same, and it thus won’t explode.

Also is there anything in there but the protein? Even if you introduced something, stuff won‘t be able to grow on just a single protease resistant protein snywsy.

Not like it‘s meant to be used for injection. Just needs to not go opaque right?

[removed]

[removed]

[removed]

If they autoclave it, a sealed vessel will still explode, as the pressure inside the vessel and the machine will still change at different rates. Plus, killing the bacteria won't make the solution less cloudy, it will just be cloudy with dead bacteria instead of living ones. And, worst of all, heat treatment will most likely kill the fluorescent proteins anyways.

[removed]

[removed]

[removed]

[removed]

[removed]

[removed]

[removed]

The dose rate seems to be very high for DU gamma radiation. I calculated 400nSv/h using 0.7 Sv/Gy on contact for 0.2 w/o U235 for newly depleted material. The dose rate will increase a bit as daughter products accumulate over thousands of years, but that isn’t particularly relevant.

>Without being replenished by new protein, the GFP would photobleach pretty quickly anyways, so I don't think the necklace is worth trying to sterilize. While GFP is stable to protease digestion, it doesn't really remain fluorescent in a purified solution for more than a few days if you just leave it our like you would with normal jewelry. Plus, killing the bacteria probably won't improve the appearance, as they likely broke down a lot of your GFP for food and their corpses will keep making the solution cloudy even if they're no longer alive. Honestly, it's probably easier to write this one off and make a new one.

I think you underestimate how much fluorescent protein we are talking about... This is milligrams. This is many orders of magnitude more protein than you are seeing photobleached in an immunofluorescence assay.

To put this in perspective, this liquid containing fluorescent protein entirely absorbs a 473nm laser I have. None of it makes it out the other side.

[removed]

Nothing but protein, but there is phosphate from the buffer as well.

Edit: An autoclave would almost certainly denature it. There is resistant to denaturating and then there is resistant to autoclave lol. Prions are one of the few proteins that are resistant to autoclaving and they are considered exceptions.

>The dose rate seems to be very high for DU gamma radiation. I calculated 400nSv/h using 0.7 Sv/Gy on contact for 0.2 w/o U235 for newly depleted material. The dose rate will increase a bit as daughter products accumulate over thousands of years, but that isn’t particularly relevant.

This is just the default calculation of cpm to microsieverts by my radiation counter (GM500) so I suppose it could be that the calculation is wrong.

Would this make UV sterilization impossible?

[removed]

[removed]

[removed]

[removed]

[removed]

Would UVC light work ?

[removed]

UV sterilization would definitely photobleach GFP in the time it took to kill the bacteria.

> Regardless, even large quantities of purified GFP need to be stored at -20C in the dark to remain fluorescent long-term, and raising that temperature to even 4C will dramatically shorten storage time. The rate of photobleaching and degradation at RT isn't going to be affected by the concentration of the protein, it just might take slightly longer to see a difference by eye. This is still a very temporary piece of jewelry, so sterilizing it at this point isn't going to extend its lifespan by much.

I'm sorry but this is simply incorrect. Photobleaching is by definition caused by light being absorbed. A high concentration of protein on the "shell" of the solution is going to "protect' all of the protein on the inside of that shell from from excitatory light. Again, I don't think you can conceptualize how much protein this is compared to "normal" amounts seen in laboratory experiments.

If I left it out in the sun, sure, it's going to bleach for in a week, but the photobleaching power of incidental lighting is just not enough to photobleach this amount of fluorescent protein any time soon.

I have tubes of nonsterile fluorescent protein that have been kept at room temperature for a year now that are cloudy (with contamination) but still fluorescent. Only my sample kept in the sun lost fluorescence.

Additional Note: this is mNeon, not GFP so it is definitely a "better" generation of fluorescent protein. But even GFP at this concentration is going to resist photobleaching for an absurdly long time.

>The "absorbtion" you see with your laser beam probably has more to do with scattering of the laser rather than pure absorption. Any high-density protein solution will behave similarly.

There is no blue light being "scattered" (I have used a blue filter I scavenged to check) , it is not scattering. You can also clearly see the beam go in, stay a beam but just fade into nothing.

>Would this make UV sterilization impossible?

Yes, unfortunately the fluorescent protein would absorb the UV and protect the bacteria! Although there is a small chance UVC might work.... That is well outside of the absorption spectra of the protein

[removed]

[removed]

How long would someone need to be cryogenically frozen for cosmic radiation to be a significant threat? If it’s a couple lifetimes then I could still see people taking that gamble (not something I’m into myself, just enjoy learning about this). Could treatment before freezing with something like iodine or anti-radiant treatment reduce that danger?

[removed]

[removed]

It could be autoclaved in a pressure cooker if the protein can tolerate the temp.

In a pressure cooker the pressure is also exerted on the outside of the ampule. No pressure differential, no kaboom.

At that point, the sample might be outputting enough energy to actually sustain life instead of killing it off lol

The gm500 has two tubes right? I don't know how it chooses which one to use (probably changes after it gets saturated), but if the standart tube is something like a sbm-20 or j305 it probably picks up quite a bit of beta that goes through the ampoule, and the cpm to usv is most likely calibrated with the energy of cesium, so i think the actual doserate is lower that what it shows

[removed]

>The gm500 has two tubes right? I don't know how it chooses which one to use (probably changes after it gets saturated), but if the standart tube is something like a sbm-20 or j305 it probably picks up quite a bit of beta that goes through the ampoule, and the cpm to usv is most likely calibrated with the energy of cesium, so i think the actual doserate is lower that what it shows

I placed whichever tube was more sensitive over the sample (one of them barely detected anything). Good call on the calibration.

[removed]

From your picture, it looks like the glass should be enough to block almost all of the radiation anyway.

A few cycles of freeze-thaw alone will kill the bacteria fairly effectively, the problem is that it may break your glass vial and damage the protein inside.

Yes, but that again requires a fairly high dose, since bacteria do have ways of dealing with ROS that can be stepped up in times of oxidative stress.

Exactly. I had a cosmology professor who rounded pi to 1 in estimating the average density of the universe or something

>A few cycles of freeze-thaw alone will kill the bacteria fairly effectively, the problem is that it may break your glass vial and damage the protein inside.

The protein will be fine. It's already been freeze thawed a bunch of times in a plastic tube. But yes, the glass breaking is a problem.

[removed]

Iodine only protects the thyroid against radioactive isotopes of iodine. That's interesting after a nuclear explosion or a major power plant accident because Iodine-131 is a significant product of them, but it doesn't do anything otherwise.

> How long would someone need to be cryogenically frozen for cosmic radiation to be a significant threat?

A short-term dose of ~100 mSv is the lowest amount where we are sure it increases the cancer risk. If we assume accumulated dose during freezing acts like a short-term dose then we need ~200-300 years to get there. Damage from the freezing/thawing process is probably still your main concern here. If we look at doses so high that they can kill you short-term then we need over 1 Sv, or thousands of years. This is assuming no special shielding in any way, and it's also ignoring terrestrial radiation sources. Normally most of the radiation dose comes from that part and people get something like 2-3 mSv/year, so we would reach 100 mSv after 30-50 years or so and over 1 Sv after a few centuries.

[removed]

[deleted]

>Glass breaks when there's a temperature differential between one area and another. > >Heated from room temp in a water bath, it probably won't happen.

OP was talking about freeze thaw. The expanding of the ice would be the problem.

[removed]

[removed]

[removed]

Keep in mind that the sample is tiny, the dose rate falls off as the distance squared, and only a small fraction of your body can be exposed to what ever the tiny Geiger counter right next to the sample tells you.

As far as bacteria etc goes they are far hardier when it comes to radiation, and to kill them takes billions times more radiation than your sample emits in an hour.

Addendum: this is how radiation oncology works. Cancer cells are constantly dividing, healthy cells aren't, so cancer cells are killed more quickly.

That's the elevator pitch, anyways.

[removed]

[removed]

[removed]

mbf explains everything well. I will only add that I heard this in an online lecture (MIT graduate course on nuclear physics I believe) though the professor said he didn't actually do the calculation to determine how long that would actually take. But I would estimate it between 10000 to 100000 years factoring in all possible background radiation sources which corresponds to mbf's estimate. So maybe it's not as crazy as I was led to believe.

[removed]

[removed]

[removed]

tbf when you're dealing with orders of magnitude a 5x difference is practically a rounding error

Radiation has become a boogeyman. It’s like UV light from the sun. We all experience it. Some places more than others. But it’s not certain cancer if you go to a place with higher radiation. It’s 0.05% more cancer if you hang around all year.

[removed]

[removed]

[removed]

Irradiation of foodstuffs is a thing. It's probably not used as much as it should be.

[removed]

Which would depend on your specific protein, and prions aren‘t that specific.

Sure globulins will be gone and other unstable protein, but since you didn‘t mention the protein.

Can just pasteurise in an autoclave as well.

No need to do 121C if you aren’t going for medical sterility. Most stuff does way earlier.

Well phosphate and protease resistant protein don‘t make up a very good growth mediums

So not much risk of colonies forming anyway.

Though got any further attempts you got stuff like propylenglykol or regular preservatives available. No use to go toxic.

Could even just use thiomersal if you still got some lying around.

But sorbic acid if acidic or parabenes if neutral to whatever if basic. That stuff works for creams that people touch daily just fine to prevent growth.

Other way round, find someone with an x ray in a lab and just use that.

[removed]

> So I mean if you wanted to wait a couple hundred thousand years...

This doesn't even work that way because it doesn't take into account the repair mechanisms of microorganisms. Too low doses will yield such low damage that it will be repaired in time.

In healthcare in America, we have a goal of sterility assurance to 10^-6 log reduction per AAMI standards but most IFU’s for devices set by manufacturers go for even tighter, some devices go beyond the 10^-9 but the trade off is damage to the device due to long exposure.

Part of the reason healthcare is so expensive is processing sterile instruments and replacing them fairly regularly, (a big push for single use disposable items is happening as a result but is also wasteful and expensive, but saves time on reprocessing)

Lastly a fun fact about steam sterilization, if you could fit the entire planet in a saturated steam sterilizer and ran it at 273 Fahrenheit for like 15 minute it would theoretically kill every thing on Earth. Nothing would survive. (Or it would be like .0000000000000001 survival)

[removed]

[removed]

[removed]

[removed]

I'm not certain, but the freezing may actually kill more bacteria in that scenario. At least most cells don't like freezing and unfreezing.

I'm not certain, but the freezing may actually kill more bacteria in that scenario. At least most cells don't like freezing and unfreezing.

Cosmology and astronomy are the only field of science where I feel that the numbers are just made up, and the measurements don't matter.

I'm not certain, but the freezing may actually kill more bacteria in that scenario. At least most cells don't like freezing and unfreezing.

I'm not certain, but the freezing may actually kill more bacteria in that scenario. At least most cells don't like freezing and unfreezing.

[removed]

Had a few classes where if we were within one order of magnitude of the correct answer it was marked close enough.

With numbers so big it’s very easy to be off by wayyy more than that

Your imaginary planet-scale autoclave would not come close to killing everything on earth. It would just turn it into a planet full of extremophiles!

[removed]

[removed]