Experimental Design:

1) Wash gametes in artificial seawater (ASW) 3 times

2) Add 1ml of diluted sperm to the gametes in ASW (Fertilize gamates)

3) Wait until more than 75% of the embryos undergo first cleavage (check for cleavage under compound microscope periodically) See Figure 1.

4) Split the embryos into 4 open dishes labeled; 1min UV, 2 min UV, 5 min UV, Control

5) Place the open dish with embryos labeled 1 min UV 3 1/2 inches under the hand held UV lamp and turn on the short wave lamp for 1 minute while wearing protective goggles

6) Repeat for the two dishes labeled, 2 min UV, 5 min UV for times indicated

7) Wait for the second cleavage to occur, take pictures with digital camera while under the microscope of the control embryos and the three experimental groups of embryos.

8) Wait for the control embryos to hatch

9) Come in the next morning look under microscope and take pictures of all 4 groups of embryos to compare stages of development and percentage of normally developed cells

Figure 1. Sea Urchin embryos during first cleavage when radiated with UV-C.