Insulin Blood Level IMPC_INS_002

Purpose

The insulin concentration in the blood is an important indicator of diabetes.

Ontological description: abnormal circulating insulin level [MP:0001560]; increased circulating insulin level [MP:0002079]; decreased circulating insulin level [MP:0002727].

Experimental Design

Minimum number of mutant animals: 7 mice for each sex.

Age of animal: 16 weeks.

Sexual dimorphism: Yes.

Equipment

  1. ELISA plate reader / MSD Sector Imager
  2. Vortex
  3. Refrigerated centrifuge
  4. Eppendorf tubes
  5. Calibrated Pipettes

Procedure

  1. Blood is collected by the relevant blood collection procedure (see IMPC protocol "Blood collection by retro-orbital puncture"). Blood is collected in lithium heparin tubes and the samples are kept on ice for a maximum of 2 hours prior to isolation of the plasma.
  2. Blood samples are centrifuged at 5,000 x g for 10 minutes at 8ºC and the plasma removed and aliquoted for analysis or for freezing (-70ºC).
  3. Plasma samples are subsequently defrosted and the required amount of sample is used to perform the analysis (e.g. by ELISA or MSD).

Notes

Blood collection for Insulin Blood Level is performed as a non-fasting, terminal procedure.

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. Plasma samples must be free of Fibrin clots in order to be analysed.
  2. Badly hemolysed samples should not be included in the analysis.

Example Metadata

Metadata

Example

Required for data upload

Required for data analysis

Type of kit

The kit used for analysing the blood samples. E.g. Mouse Insulin kit

YES

NO

Kit manufacturer

Manufacturer of the kit. E.g. MORINAGA (Yokohama, Japan)

YES

NO

Kit lot number

 

YES

NO

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.

YES

NO

Equipment manufacturer

Manufacturer of the equipment. E.g. Thermo scientific.

YES

YES

Equipment model

Model of the equipment. E.g. Multiskan JX.

YES

YES

Blood collection tubes

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

NO

YES

Anesthesia used for blood collection

The anesthetic used during blood collection. E. g. Isofluorane.

YES

YES

Method of blood collection

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

YES

YES

Anticoagulant

Anticoagulant used for blood collection. E.g. Li-Heparin.

YES

YES

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.

YES

YES

Date of measurement

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

YES

YES

Sample status

Indicate 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.

YES

YES

Samples kept on ice between collection and analysis

Yes/No.

YES

YES

Plasma dilution

Dilution 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.

YES

YES

Replicates

Please specify whether samples were measured once, in duplicate or in triplicate. E.g. 1 or 2 or 3.

YES

NO

ID of blood collection SOP

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

YES

YES

Hemolysis status

The gauged degree of hemolysis. E.g. slight/moderate/marked.

NO

YES

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.

YES

NO

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.

YES

NO

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.

NO

NO

 

Parameters & Metadata

Name Version Type Req. Upload Req. Analysis Annotation Increment Option Derived Unit Data Type
Name Version Type Req. Upload Req. Analysis Annotation Increment Option Derived Unit Data Type