Step 13 |
The emulsification mixing step does not produce a viscous solution |
Insufficient mixing |
Allow emulsification syringes to sit until the CFA and DPBS phases separate. Attempt to emulsify again. Repeat (1–3 times) until there is a noticeable and consistent shift in viscosity |
Step 22 |
The emulsion is runny and loses its viscosity when loading 1mL syringes |
The emulsion is too warm |
Place the emulsion at −20°C for a minimum of 30 min. Keep the emulsion on an ice pack while loading. |
Steps 28, 30 |
The emulsion pellets disperse following injection |
The emulsion is too warm |
Keep the 1 mL syringes on an ice pack or place at −20°C until the solution has become more viscous. |
Disease onset/Clinic al scoring |
Symptoms do not occur or disease incidence is low |
Improper preparation or faulty/expired reagents |
Double-check all calculations, concentrations, sample preparations, and reagent expiration dates (especially PTx). |
Improper MOG35–55 CFA pellet injection location |
Euthanize and dissect mouse to verify that the pellets are in proximity to the inguinal and axial lymph nodes. Adjust future injections accordingly. |