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

Project protocol: Clinical blood chemistry

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

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