I've noticed there was alot of feedback from the other post I made that I wasn't able to get to, and I'd like to say thanks for all the references and tips to get me started!  I have been so busy with finals (chemical engineering major), and organizing a group of friends into researching a variety of topics, that I haven't been able to find the time to write my thanks!!

Second, I finally got the cultures, and built a decent apparatus to grow them with, which is versatile enough to let me grow many cultures at varying light conditions etc easily and at the same time.  I am growing the algae, which is a species of Scenedesmus, in 20x150mm test tubes, and I have not seen anyone else try this method.

First, does anyone know what type of organic waste algae put off as they are growing?  Using dilution rates in larger set ups, I don't think this causes much of a problem, but evaporation from my test tubes I believe might cause a build up and negative effect on the growth.  Does anyone know how these organic wastes adversely affect cultures, or if it is not a major concern?  Also, is it a good idea to make up the evaporation losses with deionized water and then do a withdrawal and replenishing of the culture from the stock medium solution?  Or can I just make up the evaporation losses with stock solution?

Second, I am growing my cultures at around 70lux of a Hortilux Blue wide spectrum bulb, with a relatively diminished amount of yellow light that is normally unavailable to the chlorophyll a and b anyways.  Is 70 lux a good amount of light or too much?  The light is a 400 watt, so I am using distance to diminish the amount of light, but I might use neutral filters if space isn't allowing.

Third, I realized today the medium I got from UTEX has no trace nutrients in it, and I was wondering if anyone knows of a good nutrient spike or locally available (common) plant supplement that has trace nutrients like vanadium and manganese in it.  Those things are essential, and I wonder why UTEX sells medium like that.

Fourth, does anyone know of a good way of sterlizing equipment in a bench scale home lab?  Would using a basin of rubbing alcohol and then thoroughly rinsing with D/I water be sufficient?

Thanks again!!! 

I meant 7000 lux, and I recently scaled it down to 3000 lux, but it doesn't look like anyone wants to help me anyways. 

Well,  nobody helps because very few are really growing algae. And the field is not well reserched. You are in uncharted territory. So, do the best you can, experiment and create a knowledge base. I grow botryococcus braun. It growe extremely slow and there is very little data from any source. I'll just keep plugging along and develope as much data as I can. I think just keeping it alive and growing at all is a major achievement.

 There is a company in El Paso, TX that is working on this same project.  good luck, we need as many sources for bio as possible.

Davturner,

Congrats on taking the next step. Those few of us on this forum who are actually growing algae are as much in the dark as you are which is why nobody is actually answering you.

Growing algae in test tubes and flasks in the lab has been done countless times. What you need to do is find out for yourself what keeps your algae alive and kicking. As it expands extend your growth volumes by increasing the size of your containers. From test tubes, to 500 ml flasks, to 1 litre flasks, to 50 litre aquarium tanks.

If you can keep it growing and expanding you are part way to starting a commercial growing system. Manipulating the 5 major environmental issues - light, temp, ph, salinity, nutrients  - will teach you at what levels you get to achieve fastest growth with highest lipids.

Keep us posted.

Well, thanks everyone.  I hope I didn't sound cynical in expressing my wish for more help, I thought there were a few growers on here.  Slippery, do you know of a good way to scale up to a 50L aquarium tank?  It seems like your average rectangular tank is pretty deep (relative to the light source) in which only the first few cm deep of the culture would be getting good light.  Turbulence and mixing from aeration gives more than enough turbulence to keep scenedesmus afloat, but I was trying to think of a mixing device that would give a decent circular mixing pattern, almost like a larger version of the magnetic stir bar on a hot plate with magnetic stirring.  And is using a meter that displays ppt a good measurement for salinity?  Or is conductivity a better measurement?

 If I gave anyone tips on starting a set of cultures trying to use scientific methods, get your chemicals first.  Some nutrients, like your Cu2+ source, are very hard to regulate using local hydroponics supplies, or even mineral extractions from gardening nurseries.  A small miscalculation of a few milligrams in a small test tube sample will kill the whole culture fast.  For trace elements, I suggest using inorganics with a Cl anion, like CuCl2, MnCl2, etc.  Make sure it is soluble.  Also, you will probably want to get a diamond scale with milligram (0.001g) resolution, or else you'll probably kill the cultures with Cu poisoning.  I would also suggest getting a microsyringe or micropipette, so that you can use stock 1M HNO3 and NaOH readily available at hydroponic stores without diluting the acid/base down first.  This is because, in the end, you get a much smaller dilution factor of the culture in the test tube, instead of adding 10 drops of pH 1.5 solution, you can add a few microliters of pH zero solution, which makes a difference with 20mL cultures.  And it saves time.  I would also say, using a 24hr light regime is not negative for starting cultures, or in general, really you just get rid of dark respiration, which is good for fledgling cultures.  Also, make sure you filter your air, and use mostly sterile conditions, contamination sucks =/  Also, find a good pH/conductivity meter with a ppt (parts per thousand) reading on it.  pH strips from a pool store are also good for a while.

 Also, with a 2000x microscope, will I be able to see differences in chloroplasts and cell wall compositions?  I don't know whether the 50$ difference between 1200x, 1600x, and 2000x microscopes is worth it, because I want to do selective breeding later.  Thanks.

davtuner:

 Also, with a 2000x microscope  

This is the type of 2000x w/camera that I use. Cheap, easy, delivered to your door.

Davtuner, send me a PM.

we have reperatedly posted detailed information on algae culturing and expanding cultures here on this forum and even have pix of our culture lab in the pictures section of this forum as well as on our website. we sell cultures by the 50 ml tube up to multiple gallons with special orders up to 100 gallons .. go back through the archives and youll surely find our posts including a list of twelve lipid producing algae, thier mediums required.and. how to expand the cultures in to  multiple gallons so as to have enough to seed a pond or pbr.which our buyers are doing with great success.

 evryone keeps trying to reinvent the wheel...

 theres no secret on how to grow the algae we even offer seminars on how to do it and publish a comprehensive manual on the subject in which we also discuss in length  the current state of the art in oil from algae extraction and other  algae biofuel related subjects.

. were trying to decide which day in may well be holding ther next seminar. weve been holding them almost back to back lately  the last one i conducted in both english and spanish to a group of masters level  AG. scioence students from Mexico in a program sponsored by the UGA and the office of economic development through the TIES program.

 we intend on putting this technology in the hands of farmers and laymen in a manner that anyone can understand rather than spouting a bunch of scientific terms at them we reverse engineer down to the simplest level so as to  make the technology redily available to anyone so as to counter the present trend of disenfranchising american farmers because of foreign competition and providing them a practical renewable technology so farmers can make  biofuel feedstocks as well as biofuels without causing perceived shortages of strategic food crops such as soybeans and corn by using algae production and the resulting stream of value added products that result from algae production for biofuels. our approach creates new and profitable income streams for anyone wishing to get involved, some who perhaps havent had the benefits of a higher education. we also train people so that they may enter new career paths in this fast growing field.that we have been involved with for the past thirty years. in fact since we are expanding explosively we are considering creating a closely knit franchise across the nation providing technical support and marketing strategies that will ensure the success of this endeavour on a permanent basis not the like the current state of the biofuels industry which is fraught with people shooting from the hip and  making uninformed decisions such as the absolutly unexcusable situation which exists today, that is people, investing in huge biodiesel plants before  securing a full knowledge of the logistical limitations of trying to produce biofuels from food crops that are  price controlled by the commodities exchange

 this carelessness has led to having at least  90 biodiesel plants across the nation sitting fallow without feedstocks to run in them. this in great part due to poor due dillegance and in many cases just plain greed ....people grabbing the millions of dollars of grant and federal loan guaranteed money that is being carelessly thrown about and lining thier pockets  with it. then shrugging thier shoulders as they get into thier allantes and Humvees saying '''"geee , sorry it didnt work""".and then heading back to thier 750 thousand dollar homes and thier trophy wives  all paid for by our tax dollars.

marc.

 wow, 2000x and only $300? How clear are your images and how bright are they? The reason I ask is that in my lab we have a 1000x microscope that costs $30,000. Didn't know they came two orders of magnitude cheaper and still of good quality. 

 You don't need more than 1000x really. They are relatively big (10 microns). Here's a video of scenedesmus dimorphus viewed at 1000x using photocontrast. 

http://youtube.com/watch?v=AuaJDkTEzrg

 Also, you shouldn't work in the units lux/lumens. These are anthrocentric; they are meant to give an indication of brightness as perceived by humans. The brightness of a light in lumens/lux is weighted according to the one of the Luminosity functions (http://en.wikipedia.org/wiki/Luminosity_function). 

Thus,  100 W/m^2 yellow light  (550-600 nm) has many more lumens than 100W/m^2 blue light (~450 nm). Your algae don't care about how we perceive light, in other words. And since the absorption peak for Chlorophyll A is in the blue range, using lumens would be a serious mistake.  The only relevant unit is energy per unit area per unit time (W/m^2). 

Hope that helps.

Also, let us know how your experiment is progressing. I'm currently doing similar light studies on dunaliela tertiolecta. 

TrevorPrinceton:

 Also, you shouldn't work in the units lux/lumens.

IMO, the best grow lights for the $ are t-5's. There are excellent for growing Higher flora... I could imagine the same for lower flora.

TrevorPrinceton:

 wow, 2000x and only $300?

I use them everyday, sometimes 2x a day. Im happy. I notice the local U used the same exact scope, only they splurged and bought the dial tune'able platform. I would actually recommend that upgrade from the one I posted, tho you can buy it add-on/after market.

Let me ask you something... do you really need a 30,000$ scope or do you just need a scope?

For a few micron size, these scopes are more than good enough. I would send a pix but all of them are Intillectual Property. I will go grab some lake water in the next few days and post some pix.

The camera feature is cool. I dont even look into the eye pieces anymore, just at the lab laptop.

I'd like to see those pics. Just curious.

No you don't need a $30,000 scope. The one we have here is top of the line and not necessary for just viewing algae. I just had no idea there would be a 100x difference between the two scopes for the same amount of magnificaiton. No offense.  

froggy:

TrevorPrinceton:

 wow, 2000x and only $300?

I use them everyday, sometimes 2x a day. Im happy. I notice the local U used the same exact scope, only they splurged and bought the dial tune'able platform. I would actually recommend that upgrade from the one I posted, tho you can buy it add-on/after market.

Let me ask you something... do you really need a 30,000$ scope or do you just need a scope?

For a few micron size, these scopes are more than good enough. I would send a pix but all of them are Intillectual Property. I will go grab some lake water in the next few days and post some pix.

The camera feature is cool. I dont even look into the eye pieces anymore, just at the lab laptop.

OK, you convincved me. So, put it in layman terms. What is the best light? Incandescent? Florescent? 60 Watt or 100 Watt? I just want a reasonable choice to go looking for.

I am using 35watt cool white florescent tubes. I think you will find most labs use the same.

 Froggy,

The last ones you recommended were very expensive. (I couldn't find a cost on these.)

Many algae researchers use common tubes.  They are cheap and seem to work. 

Are the T5s available for low prices - like Home Depot?

liberty1:

 Froggy,

The last ones you recommended were very expensive. (I couldn't find a cost on these.)

Many algae researchers use common tubes.  They are cheap and seem to work. 

Are the T5s available for low prices - like Home Depot?

Fluorescent Lightbulbs are NOT the way to go.  They are completely inefficient since most of the light the emit cannot be absorbed by chlorophyll. Check out the emission spectrum for fluorescent lamps and then check out out the absorption spectrum for chlorophyll. The spectrum for the f. lamps peak narrowly in regions where the algae cannot use the light. They do not coincide at all. 

The reason people use fluorescent lamps is...well..they have no reason to. I don't think people have really thought hard about what they are doing. They just sort of do it.

The best light to use is probably the one whose spectrum most closely resembles the sun's

These things have evolved to use the sun's energy spectrum. You might want to find something that contains only the visible spectrum, since we know now that  UV and IR radiation are pretty much useless to living creatures aside from heating things and causing higher mutation rates. 

TrevorPrinceton:

   Fluorescent Lightbulbs are NOT the way to go.  

My t5's are cheaper to run, are cooler and do a better job of growing than does the HID MH systems I had. I switched over after getting my first t5 set and running them for a month or so.

I will be the first to confess that I dont know which light spectra are best for alga. All I know is that I have switched my higher flora indoor growth to t5's. t5;s were designed with plants in mind so they likely have a different spectra than just shoplight fluoro's.

No, they arnt as cheap as shoplights but IMO, worth the upgrade $. Maybe 5x the cost of shoplights. You can also buy HO t5's so your buying less fixture and more light/sq ft.

TrevorPrinceton:

  I'd like to see those pics. Just curious. 

Not sure how you can compare there two, unless you send me a slide so we can take pix of both scope/camera.

If you need such a pure pix, you should go to non-optical like SEM/TEM anyhow.