Developmental Biology
/Parastichopus parvimensis
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Methodology
Spawning Protocol
Each sea cucumber was injected with about 10mL of a dilution of 100mM NGLWY-amide dissolved in dH2O. Males and females were separated upon initial spawning to avoid polyspermy.
Confocal Microscopy
Larvae were first fixed with formaldehyde. DAPI was used to bind DNA to highlight nuclei. Phalloidin bound F-actin, accentuating muscles. Lastly, tubulin was highlighted by using a primary antibody made in a mouse. A secondary antibody (anti-mouse), made in a goat, was then added to amplify the primary antibody.
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All experiments and imaging occurred at Stanford University's Hopkins Marine Station located at 120 Oceanview Bld, Pacific Grove, CA 93950. In addition to conducting their own world class research, faculty teach a variety of undergraduate and graduate classes at the station. All those affiliated with the station capitalize on the rich waters of Monterey Bay.
To learn more about Hopkins Marine Station, visit their website at http://www-marine.stanford.edu/about.php