How to Obtain and Culture Marine Dinoflagellates
One of the most impressive demonstrations of bioluminescence is also the easiest. Dinoflagellates, the most common sources of bioluminescence at the sea surface, are readily grown in the laboratory. They produce bright bioluminescence upon agitation. The following instructions describe how to obtain dinoflagellate cultures and grow them at home, school, or laboratory. Then check out the Web page on bioluminescence demonstrations.
Dinoflagellate Cultures
Cultures may be obtained from (in the U.S.A.):
The University of Utah "Science in the Dark" project.
(801) 581-7730, (801) 585-5361 FAX
Pyrocystis lunula (hearty and bright)
Cost is approximately $9 per bag plus $5 for shipping.
The University of Texas Culture Collection:
Culture Collection of Algae
Department of Botany
The University of Texas at Austin
Austin, TX 78713-7640
(512) 471-4019, (512) 471-3878 FAX
Cost is $10 per culture, shipping is $7-12 for overnight delivery (recommended).
Delivery in 2 weeks.
Pyrocystis noctiluca LB2504 (large spherical cell, bright luminescence)
Pyrocystis lunula LB2166 (rapid growth, bright luminescence)
Carolina Biological Supply Co. [(800) 547-1733 for western U.S. or (800) 334-5551 in east]
Note: Carolina Biological no longer sells luminescent dinoflagellates, although they do offer glowing bacteria and dried luminescent ostracods.
Maintenance
[refer to Journal of Phycology Supplement 29 (1993) for additional information]
All you need are lights and a timer. If you use sterile media and glassware, your cultures will continue forever; every month pour about 1/4 of the culture into some new medium. If you can't maintain sterile culture conditions, the cells will last for a few weeks to a month before bacteria overgrow the culture.
- Room temperature (20-25 degC)
- Illumination with cool-white fluorescent lamps
- Light cycle (12 hr each light, dark - see below)
- Culture flasks -- Sterilized glassware if autoclave is available, otherwise use disposable tissue culture flasks
- Culture media (see below)
Growth Media
Just as your houseplants (hopefully) get fertilizer to help them grow, so too do dinoflagellates have nutritional needs. Dinoflagellates require nitrate, phosphate, trace metals, and vitamins. Growth medium is sterile, so if don't have an autoclave available, it is easier to buy already prepared media than to make your own.
Prepared Culture Media
From Carolina Biological Supply Co. [800-547-1733 for western U.S. or 800-334-5551 for Eastern U.S.]
- Alga-Gro seawater medium "for culturing marine algae"
Carolina Biological #15-3754, $10 per quart.
This is a complete ready-to-go solution. Just add dinoflagellates.
- Alga-Gro concentrated medium "enrichment for seawater"
Carolina Biological #15-3750, 12 tubes $10 (each tube makes 1 liter)
This enrichment is added to filtered seawater.
From Sigma Chemical Co. [800-325-3010 for USA/Canada, 314-771-5750 outside USA/Canada (call collect)]
- Guillard's (f/2) marine enrichment basal salt mixture without silicate
"With the macro- and micronutrients as described by Guillard (1975)."
Sigma #G4277, $9 for 10 liters.
This is a complete ready-to-go solution. Just add dinoflagellates.
- Guillard's (f/2) marine water enrichment solution without silicate (50x)
"With the major nutrients, trace metals and vitamins as described by Guillard (1975). Use at 20 ml/L of seawater. Sterile"
Sigma #G0154, $17 for 500 ml.
This enrichment is added to autoclaved natural or artificial seawater.
Do-your-own Culture Media (for the brave only, requires sensitive balance and autoclave).
Refer to Journal of Phycology Supplement 29 (1993) for recipes for ES-enriched seawater medium. There is also a recipe for artificial seawater, if required.
Day-night Light Cycle
The key to growing dinoflagellates is to give them 12 hr of fluorescent lighting to allow them to perform photosynthesis, and 12 hr of darkness during which they can bioluminesce.
- Dinoflagellates only emit bioluminescence during their night phase. Peak luminescence occurs 2-6 hr into the dark phase.
- Use a timer to control the lights, with 12 hr of light and 12 hr of dark.
- Set the timer so the lights go off 2 hr before you will observe the luminescence. For example, for an afternoon class, set the lights "off" at 11 am, with the "on" at 11 pm.
- It takes a few days for the dinoflagellates to entrain to a new light cycle.
Dinoflagellate Culture / Updated 9/10/99 / biolum@ucsd.edu
