Breeding Catts: A Photo Journey

[ATester]

Julie, sorry to hear about the combat death...in my line of work, we hear alot about them. I hope all goes as well as it can with the plans.

Something to try (of course, implying the incredible amount of patience that this requires with growth and bloom time that goes along with this kind of project), if the species traits aren't as prevalent, it would always be possible to recross the new hybrid with the species plant again...to strengthen whatever traits that are expressed but not as pronounced as before...the major drawback being though that the species plant is a wild card, so the variation among offspring will probably increase (but now we're talking plant genetics with is oft a strange world and unpredictable).

For those of you interested, we did have a good day at the lab. Working with a hood is ALOT different than the semi sterile methods that I am somewhat acustomed to. After preforming all of these little steps over and over again, it is really abvious how accidental contamination happens! Trying not to reach for something outside of the hood, and if you do, saying "oh crap!" and hosing yourself down with ethanol again. In some ways the hood is easier I think (open flasks, easier to plate the seeds...petri dishes to take a small sample of seed and look for embryos under a microscope).

And yes, there are some interesting photos...and interesting marks left behind by nitrile gloves...

[nabakov5]

ok, this is officially my favorite thread. the suspense is killing me. and the whole affair is so ambitious from my seats in the nosebleed section. good luck!!! i can't wait to see the newbie
cat

[p 1]

i have a few long posts i really ought to make, but i haven't gotten around to it, so i've been lurking until i do. but then i saw this thread, and wow! so exciting!

if you wind up with more tiny little experiaments than you know what to do with, hope i can help!

[Mehera]

There's been another combat death - a local fellow killed in Iraq, and the funeral with full military honors will be Thurs. I'm knee deep in arrangements and won't have time to post the pics quite yet. More to come!

McJ: So sorry to hear that your talents are needed in this way yet again. I think it is so amazing that you can lend your abilities to add dignity and beauty to the services for one of our lost young servicemen. I'm sure it means a lot to those who love him. Thank you.

[Piper]

Thanks for the kind words, Mehera. Army Spc. Jared Raymond was 20 - he graduated from Swampscott High School two years ago. His tank hit an IED, a homemade landmine, outside of Baghdad. These things really bum me out, but it's an honor for me to be able to serve the family. I feel as though I'm piping Jared's soul to heaven.

I'll pipe outside the church before the service, and then I'll lead the casket out at the end of the service. There will be a procession with the horse-drawn caisson from the church to the cemetary (two miles.) The Chelsea Fire Dept pipers and drummers will play during the procession. And then I'll play as the cemetary service concludes (Amazing Grace, and I'll finish with the chorus of America the Beautiful.)

Anyway, the flasking was a much more enjoyable way of spending a day and I'll sort out the pictures hopefully later today. Flasking orchid seeds is an excruciating tease for those with a compulsive-obsessive nature. The seed pod contains millions of microscopic seeds. Yet you cringe when any miss the test tube. You know, if all goes well, you'll end up with hundreds of seedlings coming out of flask. And yet when someone innocently asks if there might be any extras for gifts to forum members, what's my first instinct?

NO! They're mine - all mine! ME, ME, ME! Get your grubby mitts off!

Sure, P1, if these guys grow up big and strong, seedlings for all forum members!

McJulie

[KC C]

Our mitts aren't on them, just our prayers. Of course, the prayers have teeny tiny threads attached to our mitts.

[p 1]

McJ: So sorry to hear that your talents are needed in this way yet again. I think it is so amazing that you can lend your abilities to add dignity and beauty to the services for one of our lost young servicemen. I'm sure it means a lot to those who love him. Thank you.

holy crap, i never even saw that post o.o

now i feel like a schmuck...sorry...

[p 1]

Our mitts aren't on them, just our prayers. Of course, the prayers have teeny tiny threads attached to our mitts.

yeah, exactly ;D

[Piper]

LMAO, KC! But...ok...now to get back on track...

First an overview of what we were trying to accomplish yesterday and then a word or two about our prep.

Orchid seeds are generally sown in nutrient-enriched agar. Because bacteria and fungi love to feed on agar, flasking must occur in sterile conditions. This is why flasking is so challenging in the home. Sterile conditions don't exist. It's possible to keep flasking jars, the agar, the tools you use, and the seeds fairly germ-free, but it requires some tricks, copious amounts of bleach, and a bit of luck. Aaron, our Minister of Silly Flasking, has been fine-tuning his home flasking techniques for many months now. Hence the award of his recent title. But even he will lose an occassional flask to contamination.

Because I have friends in the Bio Dept at Dartmouth, I thought it would be fun to flask my seed capsules in a sterile hood. Aaron loved the idea: he'd get to play mad scientist and we'd greatly reduce the risk of contamination.

A hood is an enclosed work space that's sterile. Think of a really large fish tank where the front glass slides up and down. Slide the glass up a few inches, and you can slip your hands in, together with the things you want to work on. Sterile air flows from the back of the hood to the front, where it's sucked out by a fan below your wrists. This maintains a positive pressure that prevents air outside the hood from getting in.

So after you glove up, you spray your hands with ethanol and VOILA! they're sterile.

Our goal was to sterilize everything, cut the seed pods in the hood, scrape the seed into some water-filled test tubes and shake the crap out of them. The seeds will settle to the bottom of the test tube and the chaf will stay in suspension. Pour off the chaf and then spread what's left on the agar in the flasking jars. Sounds simple...

So when we arrived at Dartmouth, Aaron and I sat down and made a list of the stuff we'd need. He prepared the flasking jars ahead of time, so the agar inside them was already sterile: there were five large jelly jars. He also brought along some sterile gauze pads and distilled water. We grabbed Lisa (the greenhouse manager and co-conspirator) and went "shopping." At the med school stock room we bought: sterile petri dishes, parafilm like a stretchy wax paper for sealing around jar lids, scalpels, sterile test tubes, 20 ml syringes no needles - just the plunger part. They come in handy for all sorts of mischief and a gallon of ethanol. The stock room guy laughed at us, and rightly so. Aaron started reading off the list: 2 scalpels. The guy asked if size 21 was ok? We shrugged. Lisa was useful for translating and asking if we could see stuff. Mind you, everything came in gross increments. We got 50-100 of most of the things we needed 1 or 2 of!

The things we forgot to get were: sterile water, hydrogen peroxide, filter paper.

First pic is of the seed pods, which I had cut just as I was walking out the door for the 2 1/2 hour drive up.

Second shot is of three of the five flasking jars. You can't see the agar, but it's a couple inches deep in the bottom of the jars. The lids are on - the foil was just an added protection.

The final pic shows the results of our shopping spree. Note the large quantities of everything! Lisa lent us her box of gloves and insisted that we wear lab coats so we'd look the part... Don't worry, no one was fooled!

McJulie

[Piper]

Naw...no worries, P1! This thread is a bit mixed up between our rather hilarious attempts at science yesterday, and the very somber reality of the world we live in. And your unvarnished lust after my little plantoids!

McJulie