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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Sunday, September 4, 2011 3:02 PM
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I wonder if the shape of the bottles has anything to do with this. You can see that the algae has settled in particular areas. Just a thought.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Sunday, September 4, 2011 3:05 PM
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The thing is, it never happens like that when I started the culture. During the entire 13 days.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Sunday, September 4, 2011 3:12 PM
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Originally Posted by chuenwe
I took this picture after I split the bottom bottle. Did it only happen in the bottle on the right that wasn't full?
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Sunday, September 4, 2011 7:55 PM
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No, actually, the bottle on the right that wasn't full doesn't have that kind of long swaying brown algae on the bottle. Those look more like dusting. It is the one on the top shelf, right in the middle and has super light brown color one that has the most of that long brown algae.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Wednesday, September 7, 2011 2:31 PM
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Trying 2 different things on 2 different split atm. In the upper shelf middle bottle with the lighter color, I changed the bottle with a Sunkist soda bottle. It has straight wall all the way up. Let's see if it makes any different. On the upper shelf right most bottle, i used a 56um sieve to filter out all the crumbed up stuff. The color became much lighter. Let's see how it goes.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Wednesday, September 7, 2011 2:58 PM
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What did you sterilize that sieve with before you poured your phyto through it? I assume you used a funnel to guide the phyto into the bottle -- what did you sterilize that with before using it? Was there an intermediate container, like a bucket or something involved? If so, what did you do to sterilize it before using it? Everything must be considered a potential source of contamination. Please read Plankton Culture Manual by Hoff & Snell, pages 34-41 on Contamination and the various precautions against it. I sterilize my culture bottles and media with a microwave. I use foam stoppers, and leave them in until the very last second (literally). I use the same pipette only for dosing the F/2, and am careful not to touch it to the mouth of the jar. I take the stopper out only long enough to dose the F/2. When I split, I have both the new media jar and the dense media jar side by side, and both are still stoppered. I remove the stopper from each, and quickly pour the 500 ml of dense culture into the new media bottle, hopefully without splashing any (it all goes directly into the center of the mouth of the new jar -- the anti-drip rings on my media jars helps this a great deal). I have the new airline and its stopper ready to go, or else I quickly recap the new jar with its solid stopper, use an alcohol wipe on the old airline, and then quickly switch the two. I could take even greater precautions than I do, such as swabbing the mouths of the jars with alcohol, turning off all the fans in the room first, doing this transfer in a hood, etc., but find that the precautions that I take work well enough for me. I think it is helpful to be paranoid about contamination, especially when working with phyto species that have a reputation for being "difficult". With Nanno, you can basically "get away with murder", because it is such an easy phyto to culture, and is so strong and vigorous, it can shrug off contamination pretty easily. Isochrysis is probably a bit more susceptible to contamination, and should probably be treated with more paranoia. Just my $0.02.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Wednesday, September 7, 2011 3:07 PM
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Thanks for the tips. I got a set of tools, sieve, funnel, and bucket, just for the phyto, well, actually, just for the ISO. This is the first time doing it. I only washed the sieve and funnel in tap water. Air dry it and used it. I got that culture manual when I first started culturing nano and tet back in may. I read most of it that's related to what I need to do, but I didn't do any of the sterilization. lol. I know I should.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Thursday, September 8, 2011 10:16 PM
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Today is the 8th day after the first split. Looked at them using a microscope and the culture looks dense and lively. The one on the right most that didn't look right to begin with and I used a sieve to filter out the crumbs, there are still iso in there. Only handful of them in the view at 200x. I'm going to give up on that bottle and start a new one on the next split tomorrow.
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Friday, September 9, 2011 1:47 PM
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I just looked at my darkest culture at 100x and lot of them are not moving. Is this normal?
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Re:Culture Journal, Species: Isochrysis galbana (C-ISO)
Monday, September 12, 2011 9:07 AM
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As of yesterday, the 2 lighter color culture that started crumbing up didn't look right. Color seemed to have changed to a bit of greenish and seemed to be more clear. Looking under the microscope and there are still some iso, but I didn't want to try to rescue it. I dump them and split the darkest culture 60/40 into a new bottle and started a new culture.
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