100 µL HM solution
TAP
05 - Heavy Metal: Effects of Environmental Toxins on C. elegans WT
RECORD your heavy metal in TABLE 1.
- - - - - - - - - - - - - - - -
MS REMOVE ~3 mL DO NOT
WT
100 µL S-buffer
Disturb worms
Module III: Collection of C. elegans and Toxicity Bioassay Excerpts from Edvo-Kit #856 HM-WT HM-MS C-WT C-MS
20 µL
TAP
COUNT
WAIT
REPEAT steps 16-17 twice for counts at 2 min. and 4 min.
WT
- - - - - - - - - - - - - - - -
HM-WT
Record in Table 1.
12. Immediately, ADD 20 µL of the wild-type worms to the “HM-WT” tube. 13. MIX by tapping this tube 3-4 times. 14. Using the HM-WT small pipet, TRANSFER 1 drop of the mixture from the “HM-WT” tube to the opening of the corresponding chamber. NOTE: If correctly placed, the solution will rapidly move into the chamber by capillary ac- tion. If the solution does not move into the chamber, check that the chamber is orientated so that the triangular opening is facing upwards. 15. Quickly PLACE the chamber under a microscope and IDENTIFY an adult worm (900-1200 μm long) for observation. 16. COUNT the number of thrashes that occur over a 30-second time period. RECORD your count in Table 1. 17. WAIT 2 minutes. 18. REPEAT steps 16 and 17 twice so that you have additional counts at 2 and 4 minutes. 19. After the final count, RECORD any additional notes about the final appearance of the worms including movement, body shape, body curvature, and any clumping. Also record any general observations about the effect of each heavy metal on the worms and how these changed over time. 20. To collect data for the effects of your chosen heavy metal on the mutant strain, REPEAT steps 11 through 19 using the mutant strain and your HM-MS labeled items. RECORD your results in Table 1.
TABLE 1: Heavy Metal_____________________
TABLE 2: Control Solutio
Wild Type Thrash Count (30 sec.)
Mutant Strain Thrash Count (30 sec.)
Wild Type Thrash Count (30 sec.)
0 min.
0 min.
2 min.
2 min.
4 min.
4 min.
Wild Type Thrash Count (30 sec.) Mutant Strain Thrash Count (30 sec.) TABLE 3: Heavy Metal_____________________
61 Wild Type Thrash Count (30 sec.) TABLE 4: Heavy Metal__
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