User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/10/26/Tobacco seed chambers

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Third style chambers

Since I believed that my chambers were causing the tobacco seeds to not grow, I made new ones. The one below is made with nail polish and beeswax.

As you can see, the water did evaporate a small amount in this chamber. Evaporation would be acceptable except for the fact that I will be putting the miracle salt solution (Murashige and Skoog medium) that aids in tobacco seed growth. The movie was made over 19 hours.

The next chamber I tried was sealed completely with beeswax.

You can see that in this one, I didn't get a very good seal and the water completely evaporated. The movies were made with Larry's time lapse software and stitched together with his .png to .avi program. Unfortunately the .png to .avi program false colors images and doesn't have a setting for not false coloring so, the original images are color but, they have been flattened to gray scale from the program.

I really hate having to melt paraffin but, it looks like I may have to try this out for the next installment of chambers.

4th try

You know, the worst feeling I can have is when I spend a tremendous amount of time and energy over thinking things and then, I think of something that is ridiculously simple and stupendously easy compared to what I was doing. It really makes me feels moronic.

There are several components to the tobacco seed cells that I need in order for this experiment to allow me to sit and forget about it for weeks. Those are:

  • Must be able to image the seeds, i.e. the cell must have clear windows.
  • The cell must have a small volume so that the seeds grow upwards and not out of focus.
  • They must be easy to add material to if necessary.
  • The must be sealed air tight in order to prevent evaporation.

So, my first cells did do this but, I think the volume was too small and they didn't have any growth in them so they were worthless. My second attempt used beeswax, which I absolutely love and will never leave home with out this stuff again, worked some what but not to the level I would have liked.

And now, I give you the solution that will work. Cuvettes.

Cuvette: I have capped the cuvette with Parafilm so that there will not be any evaporation. The experiments in the microcentrifuge tubes had a lot of atmosphere in them. So does the cuvette so there will be some evaporation in the cell. I'm not sure if I should fill the whole thing up or not as this small amount of atmosphere may aid in the cell growth.  
Cuvette images: Crystal clear. As is to be expected with the cuvette windows.  

I suppose that I can get smaller volume cuvettes in order to keep all the seeds in them in the same focal plane. Nah, I'll work with what I have now. The above image is for 6 seeds total in the cuvette with tap water. I'm hopeful that they will start growing in this setup.

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