JaxCC1 project protocol for procedure: insulin blood level

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: 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

    • Isofluorane

     

    Procedure

    1. Mice are anesthetized with isofluorane.
    2. Blood is collected via retro-orbital puncture.
    3. 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.
    4. Blood samples are centrifuged at 5,000 x g for 10 minutes at 8ºC and the plasma removed and aliquoted for freezing (-70ºC).
    5. 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

    1. 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