GMC06 project protocol

Glucose levels and lipid profiles after fasting in 5 inbred founder strains of the Collaborative Cross   (2020)

German Mouse Clinic and Department of Infection Genetics, HZI
With: Hrabě de Angelis M, Fuchs H, Gailus-Durner V, Rathkolb B, Wolf E, Lengger C, Kollmus H, Schughart K




German Mouse Clinic: Phenotyping Pipeline

Select clinical chemistry parameters after fasting: 10 - 11 wks of age

 

  • Project protocol - Contents

    Workflow and sampling

    Step
    Procedure
    		 Equipment
    Data collected
    1
    		 Mice weighed Balance
    Body weight before fasting
    2 Mice fasted for 16 h - -
    3 Mice weighed Balance Body weight after fasting
    4 Mice anesthetized and blood samples taken - -
    5 Plasma prepared Centrifuge -
    6 Samples analyzed Analyzer Glucose, glycerol, total cholesterol, HDL cholesterol, non-HDL cholesterol, non-esterified fatty acids, triglyceride

    Equipment and supplies

    • Balance: Ohaus (AV213C)
    • Heparinized (lithium) sample tubes: KABE (078028), Numbrecht GERMANY
    • Centrifuge: Heraeus (Biofuge Fresco), Hanau GERMANY
    • Eppendorf tubes
    • Clinical blood chemistry analyzer: Beckman-Coulter (AU480)

    Reagents and solutions

    • Isoflurane

    Procedure: Clinical chemistry following 16-h fast

      1. Blood samples are taken from isoflurane-anesthetized mice by puncturing the retro-bulbar sinus; blood is collected into heparinized sample tubes; each tube is inverted 5 times to achieve a homogeneous distribution of the anti-coagulant.
      2. Samples collected from fasted mice are stored in a rack on ice until they are centrifuged (10 min, 5000 x g) as soon as possible.
      3. Plasma samples (neat) are transferred into Eppendorf tubes and analyzed immediately according to manufacturer's instructions.

    Data collected by investigator

    • Body weight (before and after fast)
    • Plasma glucose (GLU)
    • Plasma glycerol
    • Plasma total cholesterol (CHOL)
    • Plasma HDL cholesterol (HDL)
    • Plasma non-HDL cholesterol (non-HDL)
    • Plasma non-esterified fatty acids (NEFA)
    • Plasma triglyceride (TG)