Freezing Arrested L1s?


I’m wondering if anyone has experience freezing arrested L1s obtained after bleaching. Can this be done or is it not worth even trying?



it would be good if u refer to “Maintenance of C. elegans” written by Theresa Stiernagle ( ;D

“…Caenorhabditis elegans can be frozen and stored indefinitely in liquid nitrogen (−196 °C) (Brenner, 1974). The keys to a successful freeze are using animals at the correct stage of development, the addition of glycerol to the freezing media, and a gradual cooling to -80°C. Freshly starved young larvae (L1-L2 stage) survive freezing best. Well-fed animals, adults, eggs and dauers do not survive well. It is best to use several plates of worms that have just exhausted the E. coli OP50 lawn and that contain lots of L1-L2 animals. A 15% final volume of glycerol in the freezing solution is used. A 1°C decrease in temperature per minute is desirable during freezing. This can be achieved by placing the worms (in freezer vials) in a styrofoam container at -80°C. The styrofoam container can be either a commercial shipping box (with walls at least ¾ inch thick) or a small styrofoam box with slots for holding vials. After 12 or more hours at -80°C, the freezer vials should be transferred to their permanent freezer location for long term storage…”

well, i would suggest u can just proceed with the freezing steps right away, since L1 will be the one that survive during the recovery.