i need to measure 3 parameters weekly, by ELISA thus i need to collect blood every week. i want to sustain my mice health and avoid anemia shock, so what's the minimum amounts of blood that could be enough for measuring 3 parameters ?
Assuming you are collecting at sacrifice, you can get 1.5ml max of whole blood. This will result in 600uL max of serum. Most assays require 50uL/well neat serum which amounts to 200uL for duplicates repeated twice. Thus, you can barely run assays for 3 parameters. As per new guidelines for animal studies, sample pooling is not recommended. This mandates careful choice of the assays.
200-300 uL of blood will be enough to get 100-150uL (40-50 uL serum/plasma for each ELISA) serum from mice. and your mice will not go in anemia.
I support above comments for blood collection.
If you are going to perform ELISA in lot, why don't you go for CBA or Luminex assay for multiple analytes with only 50uL of serum/plasma.
Though Luminex assays are attractive in terms of sample amount, a reliable positive control needs to be included. These assays can be used only after proper validation.
Luminex assays for multiple analytes can be perfomed with samples of 50ul of serum/plasma as stated above. Moreover same assays recommend in general 1:2 until 1:5 dilutions. So, for some kits could be sufficient 10-25 ul of undiluted sample.
- Badges
- Science topic