Tag Archives: tobacco seeds

D2O Effects on Life, D2O1, Water Type Effects on Organism Growth

D2O1: Day 0

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Some things to note:

  • all D2O mixes are mixtures between D2O and DDW
  • SMOW (Standard Mean Ocean Water) is 15mM D2O
  • 1% D2O is 37.5 times more D2O than SMOW so figure ~562mM D2O
  • Because of an unforeseen sunburn (which probably was foreseen by someone) I didn’t get to keep on my schedule. So these were in the fridge until today, a full week after setup. I’ll keep an eye on the sprouting to make sure that everything proceeds as normal, but this may be a throw away experiment.
  • Also, recall that there was a mishap with the 20% D2O sample. There is a huge air bubble in there and that might negate this sample.

DOI: 10.15200/winn.142802.21830 provided by The Winnower, a DIY scholarly publishing platform

D2O Effects on Life, D2O1, Water Type Effects on Organism Growth

D2O effects on tobacco seeds setup (D2O1)

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Didn’t expect that one did you? The purpose of this experiment is to study a much more narrow range of D2O mixtures based on the RC experiments. We want to determine if there is some amount of D2O that maximizes growth in tobacco seeds. If there were it would be somewhere between none and 5% in my opinion, and could quite possibly be right at the natural water amount of D2O (15mM).

So this experiment will be setup exactly like the RC experiments but we’ll be tracking length with some software after the images are acquired. Here is how I setup the experiment:

  • There are 8 samples of tobacco seeds and each sample contains 35-42 seeds.
  • The 8 amounts of D2O are:
    • pure DDW (no D2O)
    • standard mean ocean water (15mM D2O)
    • 1% D2O in DDW
    • 5% D2O in DDW
    • 10% D2O in DDW
    • 20% D2O in DDW
    • 25% D2O in DDW
    • 33% D2O in DDW
  • I know the amounts are strange but I figured in following experiments I will do different amounts for a more robust data set.

Below is a table of the amount of D2O and DDW in each sample and the number of seeds in each sample as well:

And here is the setup:

  1. I began by preparing 8 15ml falcon tubes with the water required as shown in the table above. I used a 1mL, 2.5uL, and 100uL pipettemen to make sure the amounts were as accurate as physically possible (for me).
  2. I then setup 8 analyslides and poured the amount of seeds shown above into each sample chamber.
  3. I added the water from the first step to the chambers and then closed them.
  4. I then sealed the chambers with the vacuum grease and placed the samples in the fridge to synchronize their growth. I will remove them on Monday for Day 0 pictures.

It is important to note that there was a mishap with the 20% D2O in DDW sample. As I was closing the slide, about 1/3 of the water and a couple seeds leaked out (clumsy hands). I don’t anticipate much of a problem with the experiment except for the added exposure to atmosphere so at the end of the trial I will probably discount the results from that sample.

DOI: 10.15200/winn.142802.21826 provided by The Winnower, a DIY scholarly publishing platform

RCD, Water Type Effects on Organism Growth

RC7: D2O Only Setup and Day 0 pics

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The setup for this experiment was rather simple:

  • Opened 6 analyslides (see Experiments Products page above)
  • poured at least 30 seeds into each analyslide
  • added 6mL of D2O
  • closed chambers and sealed with corning vacuum grease

I’m going to refer to this experiment as RCD from now on both in titles and in the categories on the side. This will be the second to last Repeating Crumley Experiment I do, for thoroughness sake. The final experiment will be my attempt to do a few samples on moist pads to show that the seeds grew because of phenomenon other than D2O. How I plan to achieve this is another story. Preplan? Preplan…

Now on to the pics:

Since the camera setup for the analyslides will be used for a different experiment (coming soon) I’ll just be taking pictures of the seeds as a lump group (but I’ll have to take better ones than this batch). I’ll also do this every 7 days, but I’ll check more frequently than that in case there is some random sprouting.

Also it is worth noting that one of the samples is actually a sample from the last DDW experiment. It was not recorded for data because D2O had nothing to do with purified regular water, but I used it as a control. That was over a month ago and there are still no sprouts.

DOI: 10.15200/winn.142801.12144 provided by The Winnower, a DIY scholarly publishing platform

Open Notebook, Water Analysis, Water Type Effects on Organism Growth

Project planning notes via Handrite

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Here are some notes I took using handrite for android regarding my project plans. I don’t mean to advertise for anything, but I use this app ALL THE TIME and it is super helpful. Typing on my phone is annoying sometimes and Handrite allows me to use my finger to write (albeit sloppily). These notes were taken yesterday while I was meeting with Steve (on my phone) and now I have time to paste them in my notebook. So here is some background on my ramblings:

  • I would like to effectively analyze the amount of hydrogen-deuterium exchange in my samples. This is a phenomenon I’ve talked about a lot, know very little about, and stress a lot over. We tried FT-IR of water samples to see if we can determine differences in the water types. We’ve had mixed results in this regard. It boils down to the fact, that I don’t trust anything. How do I know that deuterium isn’t sneaking into my unopened bottles of DDW, or that hydrogen isn’t getting into my D2O? What about after it’s been opened? What about in my samples? If I’m going to get reliable results with yeast and e. coli I’ll need to know if I can trust commercial products.
  • NMR (nuclear magnetic resonance) seems to be a promising route to try and there may be some equipment either in the Chemistry department or Biomedical Engineering here at UNM that I’ll need to look into.
  • Mass spectrometry may be another avenue to determine precise amounts of deuterium in water.
  • Something not written but that I talked with Steve about yesterday and just remembered is that I need to talk with Sigma to find out how they measure the purity of the DDW and how reliable those measurements are long term. If something sits on a shelf for a year, how do I know it isn’t really just pure natural water?
  • I also have some stuff in my notes about the Repeating Crumley experiment. I just setup a D2O only experiment to show definitively that tobacco seeds do not grow in D2O. Previously I would only track this data for 10-15 days and Crumley went up to a month (if I remember correctly). I’ll write the setup in a minute, but I want to do 30 days so I have some pictures for my open access, self published RC paper (spoiler alert!).
  • Also to show that I’m no slouch, I want to try and repeat Crumley’s original experiment with paper towels to show that the growth of the 100% D2O seeds was due to H-D exchange and other environmental factors and not because they eventually grow in D2O. Yea I’m a badass.
  • The rest of the notes are some ramblings that are not really private but thoughts I would prefer not to elaborate on now.

Tata for now!

DDW5, Water Type Effects on Organism Growth

DDW5: Water Day 25 (Delayed)

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I took these pictures on Day 25, but haven’t had time to post them because of all the SDM work. So here they are:

One more set of pictures to take.

DDW5, Water Type Effects on Organism Growth

DDW5: Water Day 20 (Day 21)

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DDW5, Water Type Effects on Organism Growth

DDW5: Water Day 20 (pics postponed)

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I was supposed to have the day 20 pics today, but I took one picture and the camera battery died so I’ll have to get them up tomorrow. In the meantime I have another day to build a camera holder that can hold the camera vertically so I can fit the entire cell in the frame.

RCW, Water Type Effects on Organism Growth

RCW: Day 11

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I forgot to take Day 10 pictures yesterday so I’m making up for it. This is the last set of pictures for this experiment. Results will be up tomorrow and then data will be uploaded to FigShare so keep a look out.

DOI: 10.15200/winn.142802.20776 provided by The Winnower, a DIY scholarly publishing platform

DDW5, Water Type Effects on Organism Growth

DDW5: Water Day 15

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RCW, Water Type Effects on Organism Growth

RCW: Day 9

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DOI: 10.15200/winn.142802.20766 provided by The Winnower, a DIY scholarly publishing platform