{"Equipment":"
    \n\t
  1. \n\t\tClinical chemistry analyser
    2. \n\t
  2. \n\t\tVortex
    3. \n\t
  3. \n\t\tRefrigerated centrifuge
    4. \n\t
  4. \n\t\tEppendorf tubes
    5. \n\t
  5. \n\t\tPipettes (200-1000 ul)
    6. \n
\n","Procedure":"

\n\tSet up the clinical chemistry analyser and perform QC analyses of the control reagents in accordance with the equipment guidelines.

\n

\n\t  

\n

\n\tSample collection and preparation:

\n
    \n\t
  1. \n\t\tCollect the appropriate volume of blood required (160-200μl of plasma), for the clinical chemistry analyser being used for assessment, in a Sarstedt gel tube containing lithium Heparin with the relevant blood collection procedure (see IMPC protocol Blood collection by retro-orbital puncture). Time of day for collection is in the morning, starting no earlier than 07:30.
    2. \n\t
  2. \n\t\tKeep whole blood samples on ice until centrifugation. Centrifuge for 10 minutes at 5000 x g in a refrigerated centrifuge set at 8°C. If plasma samples cannot be analysed immediately, keep them in the fridge until analysis.
    3. \n\t
  3. \n\t\tAnalysis of samples is optimally done on the day of collection. When not possible the plasma samples can be stored at 2-8°C. If samples require storage for > 48 hours, freeze plasma at -20 °C in single aliquots. All samples are allowed to come to room temperature prior to analysis.
    4. \n\t
  4. \n\t\tUse plasma samples undiluted or diluted to a ratio of 1:2 with deionised water if the volume is insufficient.
    5. \n\t
  5. \n\t\tPlasma samples that were frozen or stored in the fridge should be vortexed briefly and centrifuged again at ~5000 x g for 2-3 minutes immediately prior to analysis. If necessary, remove fibrin clots using a wooden applicator.
    6. \n
\n

\n\t 

\n

\n\tAnalysis:

\n

\n\tSamples that produce results that lie outside the linear range for a specific assay have to be re-tested. In some cases it may be necessary to dilute samples with water to bring test results into range.

\n","Purpose":"

\n\tClinical chemistry determines biochemical parameters in plasma including enzymatic activity, specific substrates and electrolytes.

\n

\n\t 

\n

\n\tOntological description: MP:0001545 – blood physiology abnormalities.

\n","Experimental Design":"

\n Sex: Females only

\n

\n\tAge of animals: 28,53 and 80 weeks

\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.

\n

\n\tDilution. 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\tHemolysis. Two fields currently exist to capture metadata information about the hemolysis status in the clinical chemistry plasma samples.The first is the LIH Hemolysis severity score which can only be performed by clinics who run one of the Beckman Coulter AU-series of analysers. Such clinics are encouraged to capture and submit the hemolysis score of the LIH test in this field. Clinics who do not have an AU analyser are encouraged to use the second/alternative field which is simply titled Hemolysis.  Simply enter "slight”, “moderate”, or “marked" based on whether the sample is visibly haemolysed or not. Provision of this information is not compulsory and it is suggested that any clinic completes at least one field or the other (not both).

\n

\n\tData QC

\n
    \n\t
  1. \n\t\tPlasma samples must be free of Fibrin clots in order to be analysed.
    2. \n\t
  2. \n\t\tBadly haemolysed samples should be discarded.
    3. \n\t
  3. \n\t\tEach morning, all parameters are tested with control sera (see ESLIM_015_001_Annex_3: Controls for biochemistry on AU400). Some parameters are tested with control serum level 1 (Beckman Coulter System Reagent, ODC0003) and control serum level 2 (Beckman Coulter System Reagent, ODC0004), which consists of lyophilised human plasma with a normal and a pathological concentration. Other parameters are tested with specific controls from other suppliers.
    4. \n\t
  4. \n\t\tControls are thawed and vortexed before utilisation and loaded according to the analyser’s display. Control values must lie within the acceptable range indicated by the manufacturer, otherwise the specific tests must be recalibrated and specific measurements repeated. Controls can be stored in 200μl aliquots at -20°C for up to 1 week.
    5. \n
\n

\n\tMetadata and examples

\n

\n\t 

\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\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. Olympus Diagnostics.

\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.  AU400

\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\tAnaesthesia 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\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\tAnticoagulant

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

\n\t\t\t\t\tAnticoagulant drug used for blood collection. E.g. Li-Heparin.

\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\tSample status

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

\n\t\t\t\t\tIndicate if the sample were frozen (analysis on the same day of collection not possible) or fresh (analysis on the same day of collection). E.g Fresh/Frozen.

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

\n\t\t\t\t\tPlasma dilution

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

\n\t\t\t\t\tDilution is highly discouraged but if necessary indicate here. E.g. “No dilution” or 1:2. Note that results submitted to DCC are assumed to be already corrected for any dilutions made.

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

\n\t\t\t\t\tID of 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\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. Year, month, day.

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

\n\t\t\t\t\tHemolysis status

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

\n\t\t\t\t\tIf no AU analyser score is provided, indicate here the gauged degree of hemolysis. E.g. slight/moderate/marked.

\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 sacrified.

\n\t\t\t
\n
\n\t 
\n

\n\t 

\n"}