Category Archives: DDW4

DDW4: Day 27

I asked Koch to take pictures of the plants for me while I am out of town. Here are his notes regarding the experience:

Wasn’t sure of your camera settings. I had trouble with depth of focus, so I chose aperture priority (“A”)
and F=22, which seemed to work OK. I didn’t want to change your zoom, so I didn’t. I took pictures of bottoms
until half way through when I realized to take tops too (so two pictures each for arabidopsis). This was a
mistake, but since I don’t know if the photos are useful, I’m stopping now.

Photos are labeled TYPE CUVETTE_NUMBERS UP/DOWN and I left in the actual photo number, as follows

1 VG cuvettes 1-3
2 VG cuvettes 4-6
3 VG cuvettes 7-9
4 VG cuvettes 10-12 (DI, TAP)

5 H cuvettes 1-3
6 H cuvettes 4-6
7 H cuvettes 7-9
8 H cuvettes 10-12 DOWN (DI, TAP)
9 H cuvettes 10-12 UP (DI, TAP)

10,11 CA cuvettes 1-3 UP, DOWN
12,13 CA cuvettes 4-6 DOWN, UP
14,15 CA cuvettes 7-9 UP, DOWN
16,17 CA cuvettes 10-12 DOWN, UP (DI, TAP)

 

Thanks Koch for the help!

DDW4: Day 15

I can’t tell in the havanna seeds sample, but the virginia gold tobacco seeds in di and tap have tiny hairs. This is not alarming because I noticed this in prior trials. It is obvious that the hair growth in the ddw samples far exceeds that of the growth in these two samples.

DDW4: Day 7 cropped photos of tobacco seeds

And here are the cropped pictures of the tobacco seeds (no past photos). Also courtesy of JPEGcrops.

DDW4: Day 7 cropped photos of Arabidopsis (and some pics from DDW3)

Here are the cropped photos. I used a program called JPEGcrops to mass crop the images from day 7. It was sweet and quick. I also included (uncropped) photos of the DI and tap water samples from Trial 3 of the DDW experiment. I don’t know if the curviness is real or not so I’ll let you comment below if you think it’s real. My money is on yes!

DDW4: Day 7

Focus on the middle sample in each image.

I have to say I’m convinced. I’m convinced that the root hairs are a real phenomenon of growing in ddw. All of the tobacco seed samples (havanna and virginia gold) growing in ddw have at least one or two seeds that have root hairs. ALL. The DI and the tap water samples are barely growing hairs. There is a clear difference between these hairs and the DDW samples.

As for the arabidopsis. I don’t know. The only thing I’m noticing is that the stems are growing very crooked. So crooked that most of the seedlings have intertwined. This is tough to confirm because the DI sample barely sprouted, but the tap water sample appears to have longer persistance lengths (to borrow a word from DNA). By this I mean the angle of curvature is greater in the tap water than in all the ddw samples which tend to curve like no ones business.

I can confirm this by comparing to the previous batch of samples. The tap and di water samples had similarly longer curve radii, while the ddw (which had to start after) seeds got all tangled together.

What should I make of this?!?!?!?!?

DDW4: Day 2

CA = columbia arabidopsis

VG = virginia gold

There is at least one seed in every CA sample that has sprouted (in fact in many of them most seeds have sprouted). Not only this but I’d say overall that over the past two days, there is more growth in the DDW samples than there is in either the tap or di water samples. By more growth I mean the length of seedling (radicle).

As far as the tobacco seeds go, there are a few sprouts here and there, but they generally don’t sprout until days 3/4.

DDW4: Setup

As mentioned yesterday I setup the next iteration of the “DDW Effects on Life” experiment. I won’t have any pictures today and maybe not tomorrow because I’m in the process of rebuilding the photography station (need something a little more stable). The first few days aren’t that important anyway, because there are no noticeable phenotypical changes and the root hairs don’t show up for some time.

As for the setup, I’ve made a few changes that I think are for the better. Also, for all product details please see the experimental product page at the top. Let’s get into the nitty gritty:

  • There are 12 samples per seed type, and there are 3 seed types (2 tobacco, 1 arabidopsis). I’m using a new batch of Virginia Gold (tobacco), Havana 2000 (tobacco), and  Columbia arabidopsis. There are 10 samples of DDW, 1 sample of tap water, and 1 sample of DI water (I only have 3 cuvette racks and this is the limit that I can hold for now).
  • For this experiment I used a new bottle of DDW. The other bottle had been opened since September and it was time for a change (in case D exchange is something to worry about).
  1. Wearing gloves, I counted 36 macro cuvettes and lids. I labeled each lid as either CA, VG, or H for each plant type (12 each). 2 lids were also labeled either tap or DI to distinguish these samples from the DDW samples.
  2. No less than 5 seeds (at most 6 seeds) were poured into each cuvette. I’ve noticed that pouring the seeds from their wax paper pouches is much faster than sorting by hand with tweezers. It also prevents me from crushing the seeds with the tweezers or from contaminating them by touching them to another surface.
  3. 3ml of a water type were then added to each cuvette and sealed with the lids. To be clear, I would add seeds to every cuvette (per seed type) and then add water and seal, then move on to the next seed type.

Typically the seeds need to soak for several hours (I don’t have an exact number, but 24 hours works well enough) before they settle on the bottom of the cuvette.

Preplanning for DDW4

Normally in my DDW experiments I do 5 seeds per sample. I realized during RC4 that this may not be enough. The reasoning is that there was a seed that potentially grew mold in the sample and it looked very similar to the experiments from the DDW series that grow root hairs.

I believe that the root hairs are a real phenomenon, because during RC1, the DDW sample had 30 seeds and they all had root hairs well after I stopped recording data, as suggested by the results of the DDW sample in the first DDW experiment. But now I’m hoping to prove this conclusively or at least consistently.

Here is my plan:

  • 10 samples of DDW water in cuvettes.
  • Each sample has 5 seeds.
  • 2 controls, 1 tap water, and 1 DI water. I’m amenable to increasing the number of either of these samples or both, but think it is most important to increase the number of DDW samples.
  • Originally I was planning on doing just tobacco seeds, but I will also do this for arabidopsis seeds. I have no (good) data for the arabidopsis seeds, and having a large sample set will convince me that something either is/isn’t happening with arabidopsis.
  • I will also be doing 2 different breeds (?) of tobacco seeds: Cuban Havanna and Virginia Gold #1. The VG seeds will be a newer batch of seeds than in previous trials. I wanted to use the rest of these seeds, but had really poor results with the last experiment so I’m scrapping them in favor of the newer order.
  • I would really love to aliquot my seeds for future experiments, but don’t know a good way to do this. I have wax paper envelopes but don’t think they are viable for dealing with small numbers of seeds. I also have tons of PCR tubes, but the plastic may become charged and make it impossible to get the seeds out of the tubes. I’d really love some ideas for this.
I’m going to setup this experiment at some point tomorrow, and realize that I haven’t provided much time for the preplanning to propagate the scientific community, but hope that it generates some help in the time.
Also this would be a good time to pose the question: would it be possible to mark a notebook entry in such a way that visitors could be alerted that there is a community question? Basically I would like to be able to flag a post so that people could direct their attention to a pressing issue in the notebook and be able to provide help immediately rather than waiting for someone to stumble upon the post randomly. Hmmmm…