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

Project protocol: Hematology

• Introduction
• Study design
• Workflow
• Equipment, software and supplies
• Reagents and solutions
• Procedure
• Measurements
• Experimental and technical parameters

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

• Hematology automated analyzer: Siemens Medical Solutions Diagnostics Advia 2120 [See Experimental and technical parameters]
• Tubes containing EDTA
• Rotary mixer

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