Round 2.... - Orchid Board

gixrj18 · #1 11-11-2007, 11:56 PM

Well, the first flasking attempt was crash & burn. I don't think the seeds were ready (now that I've seen some that are), I guess the seed opened prematurely. I flasked them in a make-shift cooler with plexi-glass on top. Needless to say, this was a pain in the *** because it was really cramped....and it had no ventilation, so the glass would always fog up.
So, to avoid further frustration and contamination, I built a laminar flow cabinet.

It has a lower chamber, where the air enters through a pre-filter on the back wall.

While sitting in here, waiting to get sucked into the intake fan, the air is exposed to ultraviolet light. After the fan pulls it into the duct (on the front)

, the air is forced through a HEPA filter, as it enters the working chamber.

From here, it is immediately sucked through the exhaust filter & fan on the back wall, where it is expelled to the side of the unit. It has a 10' cord, and once plugged in, it has a light switch, fan speed control, and an extra power source (for plugging in the UV light).....all self-contained.

Just finished flasking round two.....MAN, WHAT A DIFFERENCE. It's amazing how much having the right tool for the job helps. I guess we'll see how it turns out, I'm going for round three, tomorrow!

Jerry Delaney · #2 11-12-2007, 12:46 AM

Wow, what a neat job in building the laminar flow hood. I can't tell from the picture, but do be careful about exposing yourself to the UV light for any period of time. It can give your eyes a terrible burn!! Also, remember that like fluorescent bulbs, the UV emission decreases over time so don't forget to replace the bulb once in while. As long as the HEPA filter is well sealed, it should perform just as good as the commercial ones I use to use in the lab.
Jerry

gixrj18 · #3 11-12-2007, 07:16 PM

Thanks, Jerry, it's nice to hear that from someone who has actually worked with one. I've never seen one in person....but this is how I figured was the most efficient way to build one. The UV light is usually only on when the door is closed, I just had it on for the picture. The HEPA filter is tightly fit, with rubber weather strip behind it.....I guess we'll know in a couple of days! I'm preparing for round three, tonight!

Jerry Delaney · #4 11-12-2007, 08:06 PM

Jasen, the way you have set up the hood it is more like a biocontainment cabinet than a typical laminar flow hood. The main difference is that the biocontainment is meant to protect the worker where as a typical LFH is designed to only protect the material being worked on. For your purpose, the panel with the gloves could most likely be removed and the hood would still work OK. I do not know how you are sterilizing your media, but during the heat sterilization process all of the air is evacuated from the flask/bottle. That means as the media cools the air is replaced with room air. For the best result, it would be best to allow the media to cool inside of the hood. Also, be sure to turn the hood on and let it run for a while (say 15 - 20 minutes) prior to use. This will pretty much remove airborn particulates that have migrated into the hood while it was not in use. The inside should also be wiped down at the start of this period using IPA, bleach, etc (including the gloves if you leave them in place). It has been many years since I did any flasking so I'm sure many things have changed, but it is kind of fun. No one did green pod in those days so I never tried that but working with dry seed is really rather easy once you get everything set up. Good luck on your future attempts Jasen. Let me know how things work out.
Jerry

gixrj18 · #5 11-12-2007, 08:40 PM

Since I don't have a pressure cooker yet, I have been sterilizing everything in the oven. As soon as it is done, I lock the oven until the next day. Then, I open the door long enough to tighten the lids on the jars.....then I leave everything in there until I'm ready to use it (now that I have the cabinet, I can store them in there). I try to sterilize the media a week or so ahead of time, so I can see if any fungus is in the media/or jar. When I get ready to flask, I spray the interior down thoroughly with bleach/water, and then let it run until everything dries. Then, I spray everything that goes in the box, as I'm loading it in there....and give the box one more misting. After waiting about ten minutes, I then proceeded to flask.

Gin · #6 01-09-2008, 11:02 AM

Nice cabinet ! I used a 10 gal aquarium made into a glove box it is a pain, no room, and as you said it fogs up , I am going to see what else could be used if I decide to do this again .
I have another pod but will pitch it .I messed up , found out the pod parent is a 4 N, pollen parent most likely a 2 N = 3N

according to what I read the resulting plant would not be any good for future breeding.

with round 3