JaxCC1 project protocol for procedure: clinical blood chemistry

Multisystem phenotyping of 18 Collaborative Cross strains   (2018)

The Jackson Laboratory

See also: JaxCC1 animal documentation         All protocol docs for JaxCC1


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

    • Isofluorane

     

    Procedure

    1. Mice are anesthetized with isofluorane.
    2. 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.
    3. Whole blood samples are kept on ice until centrifugation (if samples cannot be analyzed immediately, keep them in the fridge until analysis).
    4. 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