Hematology IMPC_HEM_002

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Purpose

Hematological assessment of blood determines blood cell counts (white blood cells, red blood cells, hemoglobin, and platelets) and additional hematological parameters (hematocrit, mean cell volume, mean corpuscular hemoglobin, mean cell hemoglobin concentration) can be derived using these indices. These tests will indicate abnormalities in the production of blood and its components (blood cells and hemoglobin) as well as in the associated blood-forming organs.

 

Ontological description: MP:0002429 - abnormal blood cell morphology/development.

Experimental Design

 

Equipment

  1. Hematology automated analyzers (e.g. Beckman Coulter AcT Diff , Siemens Advia 2120 or Hemavet Multispecies Hematology Analyzer HV950FS Drew Scientific, CT, U.S.A.)
  2. Rotary agitator

Procedure

Set up the hematological analyser and perform QC analyses of the control reagents in accordance with the guidelines provided by the manufacturer.  

 

Sample collection and preparation:

  1. Collect the appropriate volume of blood required for the hematology analyser being used for assessment (~200µl), in an EDTA coated tube with the relevant blood collection procedure (see IMPC protocol Blood collection by retro-orbital puncture). The time of day for collection is in the morning, starting no earlier than 07:30.
  2. Mix the blood sample on a rotary mixer immediately following collection for a minimum of 30 minutes and keep the sample at room temperature (for no more than 2 hours) pending analysis. Samples must not be frozen at this stage.
  3. Analysis of samples is optimally done on the day of collection. When not possible the blood samples can be stored at 2-8°C for up to 24 hours. Long term storage of whole blood is not recommended. All samples are allowed to come to room temperature prior to analysis.

Analysis:

  1. Perform hematological assessment of each sample including: white and red blood cell counts, hemoglobin and platelets in accordance with the analyser being used.
  2. Derive additional parameters for the sample that may be estimated from the initial assessment such as: hematocrit, mean cell volume, mean corpuscular hemoglobin and mean cell hemoglobin concentration.

Notes

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. Whole blood (for Hematology) and plasma (for Clinical Chemistry) require different collection tubes so two independent samples are required from each mouse. Dilution of blood is highly discouraged, but is allowed when the total necessary amount is not obtained. If dilution is necessary then the assays should be done in one run.

The information about the date of the experiment, that is the date when the measurement is performed, is an important parameter which is to be submitted in the Experiment xml file (dateOfExperiment="2013-02-28").

Data QC

  1. Sample must be free of blood clots in order to be analyzed.
  2. Some results from hemolysed samples should not be reported.
  3. Perform routinely and immediately prior to sample analysis:
  1. assessment of control samples with different levels of hematology phenotypes (abnormally low; normal; abnormally high).
  2. analysis of the graphical reports generated for each control level to ensure that they lie within their respective ranges.

Metadata and examples

Metadata

Example     

Equipment ID

ID of the machine used when more than 1 is used  having same model and manufacturer. E.g. machine 1, machine 2, machine Minnie, machine Mickey Mouse, etc.

Equipment manufacturer

Manufacturer of the equipment. E.g. SIEMENS.

Equipment model

Model of the equipment. E.g. ADVIA120.

Blood collection tubes

The tubes used for blood collection. E.g. Sarstedt Li-Heparin gel tubes or Kabe Labortechnik Lithium heparin coated tubes.

Method of blood collection

Concise description of the method used for blood collection. E.g. Retro-orbital puncture.

Anesthesia used for blood collection

The drug used for anaesthesia during blood collection. E. g. Isofluorane.

Anticoagulant

Anticoagulant drug used for blood collection. E.g. EDTA.

Samples kept on ice between collection and analysis?

Yes/No

Storage temperature from blood collection till measurement

E.g. 2°C

Date and time of blood collection

Time of day for collection is in the morning, starting no earlier than 07:30. E.g. Year, month, day, time.

Date of measurement

The day of blood analysis. E.g. Year, month, day.

ID for blood collection SOP

ID of the protocol followed for blood collection. Can be a center specific protocol. E.g. ESLIM_024_001

Chip card

The chip card contains the settings and thresholds that are used to calculate the numbers of cell types in a blood sample. As the blood cell sizes differ between the species, there are different thresholds for the categorization and therefore there are different chip cards for different species (mouse strains). Eg. C57BL/6 chip card.

The chip cards really look like a chip card. You put them into a slot on the haematology device and then you start measuring the haematological parameters of the corresponding blood samples.

Blood collection experimenter ID

An ID of any format to be used coherently both inside the same procedure and for all procedures indicating the experimenter who collected the blood. E.g. Harw_001, or 1/2/3.

Blood analysis experimenter ID An ID of any format to be used coherently both inside the same procedure and for all procedures indicating the experimenter who analyzed the blood. E.g. Harw_001, or 1/2/3.

Date equipment last calibrated

Most recent date in which the equipment (or any part of) used in the procedure was subject to a calibration event.

Date and time of sacrifice

The date and time when the mouse is sacrificed.

 

 

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