I use Soderhall and Smith's formulation. It's a sodium citrate/EDTA base (Soderhall and Smith 1983).
NaCl, 0.45M (FW 58.44) Sigma S5886 26.3 gm
Glucose, 0.1 M (FW 180.2) Sigma G5146 18.0 gm
Citric acid, trisodium, dihydrate, 30 mM (FW 294.1) Sigma C7254 8.8 gm
Citric acid, monohydrate, 26mM (FW 210.1) Sigma C0706 5.5 gm
EDTA, disodium salt, dihydrate, 10 mM (FW 372.2) Sigma E1644 3.7 gm
Mix until salts dissolve, takes a while.
pH 4.6 - 5.6, Filter 0.45um. Keep at 4C.
There are many options:
Durliat, M., and Vranckx, R. 1981. Action of various anticoagulants on hemolymphs of lobsters and spiny lobsters. Biological Bulletin 160: 55-68.
Durliat, M. & Vranckx, R. 1989. Relationships between plasma and hemocyte proteins in Decapoda. Comparative Biochemistry and Physiology, 92B, 595-603.
Thank you dear Jeffrey. Whats the proportion that you use for sodium citrate to hemolymph
The proportion depends on what you're planning on doing with the hemolymph. We've used 1:1 and 1:5 (one part hemolymph, 5 parts anticoagulant). There are a number of different anticoagulants that people use. You could post your question on [email protected] to get more responses. You have to be a member of crust-l to post to it, but membership is free.
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