See JAX Pipeline on the IMPC website for order of tasks for all mice tested
Project protocol: Insulin blood level
Introduction
The insulin concentration in the blood is an important indicator of diabetes. Note: Blood collection for Insulin Blood Level is performed as a non-fasting, terminal procedure.
Study design
Paradigm: Baseline study
Panel: Collaborative Cross
Number of strains: 18
Sex: Both
Age: 17-19 wks
Sample size: Six mice per sex from at least two litters
Number of cohorts: 1
Workflow
| Step |
Description |
Equipment |
Data collected |
| 1 |
Mice are anesthetized |
- |
- |
2 |
Blood is collected and plasma isolated |
Microtainer tubes, Eppendorf tubes, centrifuge |
- |
| 3 |
Insulin levels are assessed |
Imager |
Plasma insulin levels |
Equipment, software and supplies
Reagents and solutions
Procedure
- Mice are anesthetized with isofluorane.
- Blood is collected via retro-orbital puncture.
- Blood is collected in lithium heparin tubes and the samples are kept on ice for a maximum of 2 hours prior to isolation of the plasma.
- Blood samples are centrifuged at 5,000 x g for 10 minutes at 8ºC and the plasma removed and aliquoted for freezing (-70ºC).
- Plasma samples are subsequently defrosted and the required amount of sample is used to perform the analysis (2 replicates). Samples are diluted 1:2.
Data QC: Plasma samples must be free of Fibrin clots in order to be analyzed. Badly hemolyzed samples should not be included.
Measurements
-
| LIMS field name |
MPD variable name |
Description |
Units |
Series |
Primary measurement |
Series order |
MPD computed |
Calculation |
Method |
Data type |
| Insulin |
insulin |
insulin (plasma) |
pg/mL |
|
|
|
|
|
hormone quantification |
Float |
Experimental and technical parameters
| Equipment name |
Imager |
| Equipment manufacturer |
Meso Scale Discovery |
| Equipment model |
SECTOR Imager 2400 |
| Anesthesia |
Isofluorane |
| Blood collection |
Retro orbital |