Blood Collection from Mice and Hematological Analyses on Mouse Blood

Birgit Rathkolb1, Helmut Fuchs2, Valérie Gailus‐Durner2, Bernhard Aigner1, Eckhard Wolf1, Martin Hrabě de Angelis3

1 Institute of Molecular Animal Breeding and Biotechnology, Gene Center, Ludwig‐Maximilians‐Universität München, Munich, 2 Institute of Experimental Genetics, German Mouse Clinic, Helmholtz‐Zentrum München, German Research Center for Environmental Health, GmbH, Neuherberg, 3 German Research Center for Diabetes Research, Neuherberg
Publication Name:  Current Protocols in Mouse Biology
Unit Number:   
DOI:  10.1002/9780470942390.mo130054
Online Posting Date:  June, 2013
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Abstract

Basic phenotyping of inbred mouse strains and genetically modified mouse models usually includes the determination of blood‐based parameters as a diagnostic screen for genotype effects on metabolism and organ function. A broad range of analytes, including hematological parameters, can be reliably determined in mouse blood, if appropriate samples are available. Here we describe recommended techniques for blood collection from mice and the considerations that have to be taken into account to get adequate samples for hematological investigations. Furthermore, we describe established methods used in the German Mouse Clinic (GMC) to determine hematological parameters in the mouse. Curr. Protoc. Mouse Biol. 3:101‐119 © 2013 by John Wiley & Sons, Inc.

Keywords: mouse; blood sample collection; hematology

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Blood Collection from Mice by Retro‐Bulbar Puncture
  • Alternate Protocol 1: Blood Collection from Mice by Puncture of the Submandibular Vein
  • Alternate Protocol 2: Blood Collection from Mice by Puncture of the Sublingual Vein
  • Alternate Protocol 3: Blood Collection from Mice by Incision of the Lateral Tail Vein
  • Basic Protocol 2: Hematological Analysis of Mouse Blood Using the Sysmex XT 2000iV System
  • Alternate Protocol 4: Hematological Analysis of Mouse Blood Using the “Scil Vet abc” or “Scil Vet abc Plus” Animal Blood Counter
  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Blood Collection from Mice by Retro‐Bulbar Puncture

  Materials
  • Mice
  • Isoflurane
  • Oxygen or compressed air
  • Laboratory scale, convenient for body mass determination of mice (e.g., Kern 440‐47N, Precision scale; Kern & Sohn GmbH, http://www.kern‐sohn.com )
  • Isoflurane vaporizer (e.g., Penlon Sigma Delta Vaporizer, UNO Roestvaststaal BV, Zenvenaar, Netherlands)
  • Flowmeter for oxygen (O 2) and nitric oxide (N 2O) and/or compressed air (e.g., Flowmeter Type SF1, SF2, or SF3; UNO Roestvaststaal BV, http://www.unobv.com/)
  • Anesthesia induction chamber for mice (e.g., UNO Roestvaststaal BV, http://www.unobv.com)
  • Small glass capillaries (about 1 mm in diameter and 20 to 30 mm long; e.g., NeoLab Migge GmbH, http://www.neolab.de/)
  • 0.5‐ to 1.0‐ml sample tubes (e.g., Kabe Labortechnik GmbH, http://www.kabe‐labortechnik.de/): depending on the analyses planned plain tubes or heparin coated and/or EDTA coated tubes must be used (see protocols for sample analyses)

Alternate Protocol 1: Blood Collection from Mice by Puncture of the Submandibular Vein

  • Sterile 4‐ to 5‐mm lancets (available, e.g., from B. Braun Melsungen AG, http://www.bbraun.com/)
NOTE: While this method can be performed in conscious mice, we strongly recommend anesthesia both to minimize trauma to the animal and to facilitate collection by the researcher.

Alternate Protocol 2: Blood Collection from Mice by Puncture of the Sublingual Vein

  • Cotton swabs
  • 24‐G hypodermic needles
  • Additional reagents and equipment for ketamine/xylazine anesthesia of mice (Donovan and Brown, )
NOTE: For this blood collection method, we strongly recommend having two people working in tandem.

Alternate Protocol 3: Blood Collection from Mice by Incision of the Lateral Tail Vein

  • Mouse restraining device
  • Scalpel with sterile scalpel blade
  • Hematocrit capillaries or 0.5‐ to 1.0‐ml sample tubes

Basic Protocol 2: Hematological Analysis of Mouse Blood Using the Sysmex XT 2000iV System

  Materials
  • Mice
  • Quality‐control blood samples (Sysmex E‐Check high level, medium level and low level, delivered with target values and limits on CD)
  • Adapted reagents packs for the Sysmex analyzer (Sysmex):
    • Required for complete blood counts and differential white blood cell counts:
    • Cellpack (cat. no. 83400116)
    • Stromatolyser FB (cat. no. 94404613)
    • Stromatolyser 4DS (cat. no. 98417216)
    • Stromatolyser 4DL (cat. no. 98417615)
    • Sulfolyser (cat. no. 90414114)
    • For additional reticulocyte counts:
    • Retsearch II (cat. no. 98416211)
  • Cell clean solution (Sysmex cat. no. 83401621; https://www.sysmex.com/)
  • EDTA‐coated sample tubes (e.g., Kabe Labortechnik, http://www.kabe‐labortechnik.de/, EDTA 500 A or EDTA 1000 A Standrand) or capillary tubes (prefilled with Sysmex Cellpack buffer as diluent) and 50‐µl end‐to‐end capillaries (Sysmex Deutschland GmbH, cat. no. 99940020, https://www.sysmex.com/)
  • Overhead rotary agitator for small sample tubes (e.g., Multi RS‐60; BioSan, http://www.biosan.lv/)
  • Sysmex XT 2000iV analyzer (Sysmex, https://www.sysmex.com/)
  • Additional reagents and equipment for blood collection (see protocol 1 and Alternate Protocols protocol 21 protocol 43)

Alternate Protocol 4: Hematological Analysis of Mouse Blood Using the “Scil Vet abc” or “Scil Vet abc Plus” Animal Blood Counter

  Materials
  • Mice
  • Quality control blood samples (Minotrol 16, Control H–high level–and Control L–low level; Scil Animal Care Company, http://www.scilvet.nl)
  • EDTA‐coated sample tubes (e.g., from Kabe Labortechnik GmbH, http://www.kabe‐labortechnik.de/, EDTA 500 A or EDTA 1000 A Standrand)
  • Over‐head rotary agitator for small sample tubes, e.g., “Multi RS‐60” (BioSan, http://www.biosan.lv/)
  • “Scil Vet abc” or “Scil Vet abc Plus” (Scil Animal Care Company, http://www.scilvet.nl), including an Abc‐reagent‐pack, which contains all necessary reagents, and a refuse bag for the waste liquids
  • Minoclair solution (Scil Animal Care Company, http://www.scilvet.nl)
  • Additional reagents and equipment for blood collection (see protocol 1 and Alternate Protocols protocol 21 protocol 43)
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Figures

Videos

Literature Cited

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