See JAX Pipeline on the IMPC website for order of tasks for all mice tested
Project protocol: Clinical blood chemistry
Introduction
Clinical chemistry determines biochemical parameters in plasma including enzymatic activity, specific substrates and electrolytes. Note: Blood collection for clinical chemistry and hematology is performed as a non-fasting, terminal procedure, with some mice being used for subsequent gross pathology and other clinic-specific parameters included in terminal assessments.
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 |
- |
- |
1 |
Blood collected and plasma prepared |
Microtainer tubes, Eppendorf tubes, centrifuge |
- |
2 |
Analyze samples |
Clinical chemistry analyzer |
Electrolytes, iron, enzymes, bilirubin, blood urea nitrogen, albumin, total protein, creatinine, carbon dioxide, glucose, lipid levels |
Equipment, software and supplies
- Clinical chemistry analyzer: Beckman Coulter AU 680 [See Experimental and technical parameters]
- Refrigerated centrifuge
- Microtainer tubes containing Lithium Heparin (BD Microtainer Lithium Heparin/PST Gel Blood Tube)
- Eppendorf tubes
- Pipettes
Reagents and solutions
Procedure
- Mice are anesthetized with isofluorane.
- Blood is collected via retro-orbital puncture into a microtainer tube containing Lithium Heparin (500 µL of blood is necessary to yield at least 260 µL of plasma). Time of day for collection is morning not before 0730h.
- Whole blood samples are kept on ice until centrifugation (if samples cannot be analyzed immediately, keep them in the fridge until analysis).
- Analysis of samples is done on the day of collection without diluting the samples. Remove fibrin clots with a wood applicator stick if necessary.
Data QC: Plasma samples must be free of fibrin clots in order to be analyzed. Each morning, all parameters are tested with control sera. Controls are thawed and vortexed before utilization and loaded according to the analyzer's display. Control values must lie within the acceptable range indicated by the manufacturer.
Measurements
LIMS field name |
MPD variable name |
Description |
Units |
Series |
Primary measurement |
Series order |
MPD computed |
Calculation |
Method |
Data type |
Sodium |
sodium |
sodium (plasma Na) |
mmol/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Potassium |
potassium |
potassium (plasma K) |
mmol/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Chloride |
chloride |
chloride (plasma Cl) |
mmol/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Urea |
urea |
blood urea nitrogen (BUN) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Total protein |
total_protein |
total protein (plasma TP) |
g/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Albumin |
albumin |
albumin (plasma Alb) |
g/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Total bilirubin |
TBIL |
total bilirubin (plasma TBIL) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Calcium |
calcium |
calcium (plasma Ca) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Phosphorus |
phosphate |
phosphorus (plasma phosphate) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Iron |
iron |
iron (plasma Fe) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Aspartate aminotransferase |
AST |
aspartate transaminase (plasma AST) |
IU/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Alanine aminotransferase |
ALT |
alanine transaminase (plasma ALT) |
IU/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Alkaline phosphatase |
ALP |
alkaline phosphatase (plasma ALP) |
IU/L |
|
|
|
|
|
clinical blood chemistry |
Float |
AST/ALT ratio |
AST_ALT_ratio |
aspartate transaminase to alanine transaminase ratio (plasma AST:ALT) |
ratio |
|
|
|
|
|
clinical blood chemistry |
Float |
Total cholesterol |
CHOL |
total cholesterol (plasma CHOL) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
HDL cholesterol |
HDL |
HDL cholesterol (plasma HDL) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
HDLD/TCHOL ratio |
HDL_total_ratio |
HDL cholesterol to total cholesterol ratio |
ratio |
|
|
|
|
|
clinical blood chemistry |
Float |
Triglycerides |
TG |
triglyceride (plasma TG, no fast) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Glucose |
GLU |
glucose (plasma GLU, no fast) |
mg/dL |
|
|
|
|
|
clinical blood chemistry |
Float |
Free fatty acids |
NEFA |
non-esterified fatty acids (plasma NEFA) |
mmol/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Carbon dioxide |
carbon_dioxide |
carbon dioxide (plasma CO2) |
mmol/L |
|
|
|
|
|
clinical blood chemistry |
Float |
Experimental and technical parameters
Equipment name |
Clinical chemistry analyzer |
Equipment manufacturer |
Beckman Coulter |
Equipment model |
AU 680 |
Fasted |
No |
Anesthesia |
Isofluorane |
Blood collection |
Retro-orbital |
Blood collection tubes |
BD Microtainer Lithium Heparin/PST Gel Blood Tube |
Fresh/frozen samples |
Fresh |
Dilution |
No |
|