{"Equipment":"
    \n\t
  1. \n\t\tHematology automated analyzers (e.g. Beckman Coulter AcT Diff , Siemens Advia 2120 or Hemavet Multispecies Hematology Analyzer HV950FS Drew Scientific, CT, U.S.A.)
    2. \n\t
  2. \n\t\tRotary agitator
    3. \n
\n","Procedure":"

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

\n

\n\t 

\n

\n\tSample collection and preparation:

\n
    \n\t
  1. \n\t\tCollect 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. \n\t
  2. \n\t\tMix 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. \n\t
  3. \n\t\tAnalysis 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.
    4. \n
\n

\n\tAnalysis:

\n
    \n\t
  1. \n\t\tPerform hematological assessment of each sample including: white and red blood cell counts, hemoglobin and platelets in accordance with the analyser being used.
    2. \n\t
  2. \n\t\tDerive 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.
    3. \n
\n","Purpose":"

\n\tHematological 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.

\n

\n\t 

\n

\n\tOntological description: MP:0002429 - abnormal blood cell morphology/development.

\n","Experimental Design":"

\n\tMinimum number of mutant animals: 7 mice for each sex.

\n

\n\tAge of animals: 16 weeks (fixed). 

\n

\n\tSexual dimorphism: yes for some of the parameters.

\n","Notes":"

\n\tBlood 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.

\n

\n\tData QC

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

\n\tMetadata and examples

\n\n\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\t\n\t\t\t\n\t\t\t\n\t\t\n\t\n
\n\t\t\t\t

\n\t\t\t\t\tMetadata

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tExample     

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tEquipment ID

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tID 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.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tEquipment manufacturer

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tManufacturer of the equipment. E.g. SIEMENS.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tEquipment model

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tModel of the equipment. E.g. ADVIA120.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tBlood collection tubes

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tThe tubes used for blood collection. E.g. Sarstedt Li-Heparin gel tubes or Kabe Labortechnik Lithium heparin coated tubes.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tMethod of blood collection

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tConcise description of the method used for blood collection. E.g. Retro-orbital puncture.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tAnesthesia used for blood collection

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tThe drug used for anaesthesia during blood collection. E. g. Isofluorane.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tAnticoagulant

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tAnticoagulant drug used for blood collection. E.g. EDTA.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tSamples kept on ice between collection and analysis?

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tYes/No

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tStorage temperature from blood collection till measurement

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tE.g. 2°C

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tDate and time of blood collection

\n\t\t\t		\n\t\t\t\t

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

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tDate of measurement

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tThe day of blood analysis. E.g. Year, month, day, time

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tID for blood collection SOP

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tID of the protocol followed for blood collection. Can be a center specific protocol. E.g. ESLIM_024_001

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tChip card

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tThe 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.

\n\t\t\t\t

\n\t\t\t\t\tThe 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.

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tBlood collection experimenter ID

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tAn 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.

\n\t\t\t
\n\t\t\t\tBlood analysis experimenter ID\n\t\t\t\tAn 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.
\n\t\t\t\t

\n\t\t\t\t\tDate equipment last calibrated

\n\t\t\t		\n\t\t\t\t

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

\n\t\t\t
\n\t\t\t\t

\n\t\t\t\t\tDate and time of sacrifice

\n\t\t\t		\n\t\t\t\t

\n\t\t\t\t\tThe date and time when the mouse is sacrificed.

\n\t\t\t
\n
\n\t 
\n

\n\t 

\n"}