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Culture Journal, Species: [Isochrysis galbana]
Thursday, June 6, 2013 3:13 AM
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Culturing Journal DataSheet This first post should be updated regularly to include new information as events take place or changes are made to your system General Species: Species description: Culture source (Local): If algae, CCMP # (Optional ): http://ccmp.bigelow.edu/ Culture Establishment Date: 31/5/2013 Continuation Date: 6/6/2013 Culturing Vessel Details Salinity: 1.020 - 1.022 Temperature: 24-26 deg C pH: 8.5 Vessel description: First established in 1.5L plastic bottle, then moved to 2x 1L glass vessels Lighting description: T5 tubes x2 (24W) Lighting cycle: 16 light - 8 dark Aeration description: Moderate, near boiling Methodologies Split methodology: culture was split on the 5th or 6th day or when it gets to a darkish milk chocolate colour, splitting it before it gets too dark to avoid risking a crash. the culture was split to by a 50% ratio, (will probably do 1:3 later to increase production speed) The new culture water was microwaved for 3 minutes or till its just about to boil then left to cool before used. glass vessels and tubing were left in boiling water before cooled to room temperature. Lids are covered with plastic film and a small hole is left for air to escape. Culture medium description: 10 drops of FAF microalgae grow per 1L culture. Cell count: (if known) Reference links: Additional Information (No Pictures or Videos in the Section Please) Notes: This is my second attempt at ISO, the 1st time was a total failure with the FAF disc, it just never coloured up. This starter came from a local store and it came in a light color, altho it eventually darkened up to the ideal color. You will be required to provide photographic evidence and as much detail as possible about your project in this thread. If your thread does not contain detailed enough photos and information the MBI Council will not be able to approve your reports.
<message edited by Tomngangan on Thursday, June 6, 2013 3:59 AM>
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 6, 2013 4:01 AM
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Here are some additional info and how the culture "came up" This is day 2 day 4 This is when the color really darkened and it went dark really fast, im guessing it was at its exp growth rate stage. day 6 This is when i decided to split it, originally i wanted to wait 1 more day. but i didnt want to risk it crashing. i moved it to a glass vessel to get a more clean photo of its color. It was almost black when not put in front of sunlight or the light source... also checking it under a microscope just for fun, (heard that ISO cells are suppose to be slightly greenish colour if its active, it does confirm on the microscope, but please point out if anyone think they arent ISO i did have a video of it but i just cant find it at the moment, the cells are motile but no way a much as Tet... and i think only 80% of the cells are motile, im not sure if its a bad sign for the 20% =( This is my 1st post here on MBI.. so... if i miss put anything.. please let me know haha....
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 2:15 AM
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This is the most dense culture so far. It was about 5 days after my last split and it seems this is going ok ~ Although i have to add that somethings with ISO that it just crashes without much of a reason, 2 bottles split using same water and same condition can have very different results. im currently having a 1/3 chance of lossing a culture after every split... going to try sterilize even more and see if thats the problem.. also going to try adding silica to some culture to test the difference ^^ hopefully better results and more constant.
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 2:30 AM
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You could most certainly go to more like a 4:1 or a 5:1 dilution instead of the overly conservative 2:1 or 3:1 you are currently doing when splitting. You might also consider splitting before it gets quite as dark as you have demonstrated -- more like when it is a very dark orange, but not yet brown. Microwaving is a good sterilization procedure, but you need to make sure that it gets up to about 185F / 85C. Too little microwaving may be an ineffective sterilization procedure, and this may be the cause of your crashes. Borosilicate glass containers hold up well to this microwaving. PET (read: plastic water/soda bottles) do not hold up so well under microwaving to this temp.
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 2:50 AM
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oh ok thz jim, i normally microwave the water for about 2 to 3 mins at 1000W, which nearly push it to boiling point. and yes.. i do split the bottle early than the DRAK DRAK brown colour... the above bottle was to try out how dense it can get haha... but its a good point to split it when its still orange. Thz if i split it at 1:4 ratio... will it work better?? i was afraid that too diluted may cause crashes??
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 3:00 AM
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I microwave my Isochyrsis media for 22 minutes and 22 seconds to get 2 x 1.6 liters up to 185F / 85C using a 1100 watt microwave. 2 to 3 minutes at 1000 W is useless for anything more than roughly 1 x 400 mL. It appears that the media is "boiling" long before the media actually gets to anything close to actual "boiling point" temperature. You need to actually measure the temperature of the media. I use a very nice Fluke infrared meter that my lovely wife gave me as a gift for Christmas a couple of years ago. You can also use more conventional alcohol or mercury based bulb thermometers -- they work quite nicely, as long as they are designed for the correct temperature range. Regarding dilution, as long as you don't go lower than 10:1 you'll be fine, assuming your sterilization of your media is sufficient. According to the Plankton Culture Manual (Hoff & Snell), you are good as long as you don't go below a 100:1 dilution. I prefer to be more conservative than that, so I consider 10:1 to be a practical dilution limit for the casual hobbyist.
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 3:09 AM
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Oh... 5:1 is a pretty good splitting ratio then .. cos i have many people to supply my plankton to haha... OK~ i will microwave the media alot longer then... do you check the salinity again after it cools? cos i suppose there will be quite a bit of evaporation right? out of interest question tho... does it work if I have a kettle boiling water up, let the water cool a bit and then mixing salt and F2 with that? then the temperature is for sure near 100?
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 3:22 AM
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 3:39 AM
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nicely explained. ThZ
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Re:Culture Journal, Species: [Isochrysis galbana]
Thursday, June 20, 2013 3:49 AM
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This is my sub-station for the cultures, (ran out of space at my main station ) (the green culture in the middle is actually Tetra, which i have no where else to put at the time, and i figured im fine with it if it "contaminate" one of the ISO.)
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