Seed floats in water -- is it chaff?

[JB_Orchidguy]

I don't know why I said 3 and 3 LOL duno what I was thinking. I ment 3 and 7. OOps that would havebeen a f up. Cool will have to think about the book too. Thanks eveyrone.

[JB_Orchidguy]

This is an excert from 'Orchids from seed for pennies in the Kitchen'

"Take a small vial and fill it 1/6 with 30% Clorox solution and drop in the seed you intend to sow, then shake it to mix thoroughly. Shake it twice more over a period of three minutes, then fill the vial with boiled (NOT still hot!) rain or RO water. Dip the coffee filter in Clorox solution or spray it thoroughly with the 50% Clorox solution, place it over the small jar and depress the center, then pour the seeds into it slowly, so it doesn't run over and lose the seeds."


I didn't plan on doing it this way, but the solutions were going to be the same. I had planned to sterilize the seeds using the solutions then when the magority of the seed sunk to the bottom then I was going to use a syringe and pull off most of the clorox solution. Then I was going to add sterile water to the solution and agitate again for about 2 min then suction off the water again. Then rinse again then finaly suction down anf fill with 10 cc of water this final 10 cc of water is going to ride with the seeds into the flasks. Once the final 10 cc of water is added then I was going to suction up all the seeds and the water into a 12 cc syringe. Shake the syringe up to dispurse the seeds in the water and inject 1 cc into each flask. This is not a definate count, but I figured it would thin the seeds out to not warent a reflask, and also the water would aid to dispurse the seeds on to of the ager mix. This method is similar to making mushroom flasks, and I figured it would work with orchids as well. Just would possibly need a 16 to 14 gauge needle possibly bigger to accomplish.

Now I'm game for not using that high of a solution, but if it doesn't hurt the seeds then why mess with it? Isn't it better to be safe than sorry? But I am no means an expert, and value the totelage of others who have more experiance. Thanks everyone.

[Kyle]

I use a system simular to what your planning, except I only rinse once, and I don't, measure my quantities. Make sure you use sterilized rinse water. Autoclave it. I attribute most of my contamination problems to the rinsing step.

The problem with not replating is that you will find it difficult to have your seeds evenly spaced on the media. You'll have seeds all over, in bunches and on the walls of the jar. Hope it works for you.

I have some questions about the media from the orchids for pennies book. I understand its cornstarch based, is there another source of food (like sugar) and what provides the gelling, or is it a liquid media?

Kyle

[JB_Orchidguy]

oh I didn't plan on using that media either. From the confesions of the aurthor its a slow growing media. And you say one rinse is enough? THen I will only do one. I didn't want to leave to much clorox. Also I will be grabbing a bottom of sterile water from work. A litter of strile water should last me a little while. We go through those like crazy at work so one bottle will not be missed. And your right its not an exact way to measure the seed count, and I was hoping to use the water to spread the seeds. I figure it wouldnt hurt to experiment. I was planning on ordering from OSB and many peole said you get a generous ammount of seeds.

[JB_Orchidguy]

I asked some folks on another orchid forum I frequent on how he sterilizes his seeds. This is a cut and paist from his post.

I use Hydrogen Peroxide to sterilize my dry seed...works great. To do so, I put the dry seed in a test tube and add the hydrogen peroxide right from the bottle from the drugstore. To this I add a tiny smidgen of Dawn or Joy dishwashing detergent (or Tween if you've got it)...just enough to break surface tension. I cap the tube and shake it violently for a minute or two. Then I place the test tube in bright sunlight or directly under a very bright high intensity lamp such as a 500 watt halogen lamp, etc (watch the heat). In the bright light the hydrogen peroxide begins to degrade, exemplified by the fizzing of oxygen bubbles, and what is left behind is sterile orchid seed suspended in pure water. Then I sow the water-seed directly onto the surface of my mother plates. I have pre-treated the dry seed with a very dilute sugar solution for 24 hours before sowing, and havn't noticed any difference...the hydrogen peroxide gives me excellent disinfection without killing any seed the way Clorox does. The problem with Clorox is you have to quickly wash it all off after a few minutes. This opens the opportunity for re-infection and it also puts you under pressure of a time constraint. With hydrogen peroxide, once the bubbles are gone, you can leave the seeds in solution as long as you like...days or weeks if you must. This is nice...just in case you get interrupted during your flasking process...and you can disinfect all your seeds first and then sow your flasks rapid-fire.

Edit> I forgot to mention not to cap the test tube too tightly, the oxygen coming out of solution of the Hydrogen Peroxide will build up pressure and pop the cap right off. (voice of experience) I poke a pin-hole in the cap to prevent that.

I just thought I would pass that on. This is the method I plan on using. Going to do my flasking after I finish my finals. Thats when the school will allow me to use the lab to do it. Thanks everyone!!!