JaxCC1 project protocol for procedure: hematology

Multisystem phenotyping of 18 Collaborative Cross strains   (2018)

The Jackson Laboratory

       


See JAX Pipeline on the IMPC website for order of tasks for all mice tested

Project protocol: Hematology



Introduction

Hematological assessment of blood determines blood cell counts and additional hematological parameters. These tests will indicate abnormalities in the production of blood and its components as well as in the associated blood-forming organs. 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 - -

    2

    		 Blood collected and prepared for analysis EDTA-coated tubes -
    3 Samples analyzed Hematology automated analyzer Blood cell counts, red cell parameters, hemoglobin, hematocrit, platelet parameters


    Equipment, software and supplies


    Reagents and solutions

    • Isofluorane

     

    Procedure

    1. Mice are anesthetized with isofluorane.
    2. Blood is collected via retro-orbital puncture into a tube coated with EDTA (200 µL of blood is needed for analysis). Blood collection is in the morning starting no earlier than 0730h.
    3. Samples are mixed on a rotary mixer immediately following collection for a minimum of 30 min (keep at room temperature for no more than 2h; samples must not be frozen). Analysis is optimally done on the day of collection. When not possible, the samples can be stored at 2-8°C for up to 24h. Samples are allowed to come to room temperature prior to analysis.
    4. Samples are analyzed.

     

    Data QC: Samples must be free of blood clots. Hemolyzed samples should not be run. Run controls prior to analysis to make sure they are in the correct range.

    Formulas and derivations: Local data processing is implemented to calculate derived measures from these data, and a table of the derived measures and raw data are reported per mouse in JaxLIMS.

     


    Measurements

    LIMS field name MPD variable name Description Units Series Primary measurement Series order MPD computed Calculation Method Data type
    White blood cell count WBC white blood cell count (WBC; units per volume x 10^3) n/µL           hematology Float

    Red blood cell count

    		 RBC red blood cell count (RBC; per volume x 10^6) n/µL           hematology Float
    Hemoglobin cHGB calculated hemoglobin g/dL           hematology Float
    Hematocrit HCT hematocrit (HCT) %           hematology Float
    Mean cell volume MCV mean RBC corpuscular volume (MCV) fL           hematology Float
    Mean corpuscular hemoglobin CHr mean corpuscular hemoglobin (CHr) pg           hematology Float
    Mean cell hemoglobin concentration CHCM RBC corpuscular hemoglobin concentration mean (CHCM) g/dL           hematology Float
    Platelets count PLT platelet count (PLT; units per volume x 10^3) n/µL           hematology Float
    Red cell distr width RDW RBC corpuscular distribution width (RDW) %           hematology Float
    Hemoglobin concentration distr. width HDW hemoglobin concentration distribution width (HDW) g/dL           hematology Float
    Mean platelet volume MPV mean platelet volume (MVP) fL           hematology Float
    Neutrophils pct_NEUT neutrophil differential (percent of total WBC) %           hematology Float
    Lymphocytes

    pct_LYM

    		 lymphocyte differential (percent of total WBC) %           hematology Float
    Monocytes pct_MONO monocyte differential (percent of total WBC) %           hematology Float
    Eosinophils pct_EOS eosinophil differential (percent of total WBC) %           hematology Float
    Pct. large unstained cells pct_LUC large unstained cells (percent of total WBC) %           hematology Float
    Basophils pct_BASO basophil differential (percent of total WBC) %           hematology Float
    Reticulocytes Retic reticulocyte count (Retic; units per volume x 10^9) n/µL           hematology Float
    Percent retic pct_Retic reticulocyte differential (percent of total RBC) %           hematology Float
    MCHC   mean RBC corpuscular hemoglobin concentration (MCHC) g/dL           hematology Float
    MCH MCH mean RBC corpuscular hemoglobin (MCH) pg           hematology Float
    Measured hemoglobin mHGB measured hemoglobin (mHGB) g/dL           hematology Float


    Experimental and technical parameters

    Equipment name Hematology automated analyzer
    Equipment manufacturer Siemens Medical Solutions Diagnostics
    Equipment model Advia 2120
    Anesthesia Isofluorane
    Blood collection Retro orbital
    Samples kept on ice Yes