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Biomethodology Of The Mouse. Office of Laboratory Animal Care University of Tennessee, Knoxville. General Behavior. Nocturnal Non-aggressive towards humans Social Barbering is common Males are more likely to fight if housed with non-littermates. Weight Gain Chart. C57BL/6. Reproduction.

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Biomethodology Of The Mouse

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biomethodology of the mouse

Biomethodology Of The Mouse

Office of Laboratory Animal Care

University of Tennessee, Knoxville

general behavior
General Behavior
  • Nocturnal
  • Non-aggressive towards humans
  • Social
  • Barbering is common
  • Males are more likely to fight if housed with non-littermates
  • Estrus Cycle 4–5 d
  • Gestation 19-21 d
  • Litter Size 10-12 pups
  • Eyes open 14 d
  • Weaning 21 d
  • Postpartum estrus
  • Reproductive life ~8 mo
  • Newborns have a subtle difference in anogenital distance
  • At 9 days of age, nipples are evident in the female and absent in the male
  • Adults have a marked difference in anogenital distance
  • Adequate housing should provide the following:
    • Clean, dry and safe area with adequate ventilation, food and water
    • Visualization by personnel
    • Sufficient space to turn around and make normal postural movements
enclosure recommendations
Enclosure Recommendations
  • Primary enclosure space recommendations per the Guide for the Care and Use of Laboratory Animals
  • UTK Mouse Cage Density Policy
primary enclosure
Primary Enclosure
  • Cage Bottom
    • Easy visualization
    • Solid bottom flooring
    • Bedding and enrichment
  • Microisolator top
    • Reduces spread of pathogens
  • Wire-bar lid
    • Water bottle
    • Feed
environmental conditions
Environmental Conditions
  • Temperature 68 – 79⁰F
  • Humidity 30 -70%
  • Ventilation 10 -15 air changes/hr.
  • Noise ≥85 db can cause
    • Stress
    • Metabolic changes
    • Reduced fertility

Room Recommendations

  • Ear Tags
  • Ear Punches
  • Institutional Animal Care and Use Committee (IACUC) protocol describes all procedures that can be performed
  • A procedure is defined as any activity carried out on a mouse such as:
    • Behavioral observation
    • Venipuncture
    • Surgery
manual restraint
Manual Restraint
  • Lift mouse by the base of the tail and place on wire bar lid
  • Push the mouse against the wire bar to prevent escape and advance the hand toward the head.
  • Grasp the scruff of the neck
  • Tuck tail between finger and palm
manual restraint14
Manual Restraint
  • Click to Watch Video
mechanical restraint
Mechanical Restraint


Plastic Adjustable Restrainer

blood collection
Blood Collection
  • Survival Procedures
    • Orbital sinus
    • Tail vein prick
    • Facial vein
  • Non-Survival Procedures
    • Cardiac puncture
    • Cranial vena cava puncture
    • Axillary cut down
blood collection17
Blood Collection
  • Blood Collection Guidelines
    • Single blood draw
      • ≤1.0 ml per 100 grams of body weight
    • Multiple blood draws
      • Maximum of 1.5 ml per 100 grams of body weight within a 2 week period
    • Note: Average mouse body weight is ~20 grams but can be highly variable between stock/strain
blood collection18
Blood Collection
  • Orbital sinus
    • For collecting up to 0.2 ml of blood
    • Anesthetize mouse
    • Hold the head steady
    • Insert pipette in the medial canthus of the eye
    • Rotate the tube between thumb and finger
    • Keeping eyelids closed, apply direct pressure using gauze for hemostasis
blood collection19
Blood Collection
  • Tail Nick
    • For collecting up to 0.2 ml of blood
    • Warm tail to dilate blood vessel
    • Use proper restraint
    • Prep with 70% alcohol
    • Use #11 scalpel blade or needle to nick the lateral tail vein
    • Apply direct pressure for hemostasis
blood collection20
Blood Collection
  • Facial Vein
    • Hold the mouse securely
    • Locate the puncture site slightly caudal to the freckle
    • Apply petroleum based lubricant to the site
    • Use a lancet or 18 gauge needle to puncture the skin
    • Collect blood
    • Apply direct pressure for hemostasis
blood collection21
Blood Collection
  • Cardiac
    • For collecting up to 1.0 ml of blood
    • Anesthetize mouse
    • Insert ≤25 gauge needle under sternum at a 20⁰ angle
    • Aspirate slowly
    • Euthanize mouse
blood collection22
Blood Collection
  • Click to Watch Video
tissue collection
Tissue Collection
  • Tail Biopsy
    • Limited to a maximum of 2 times
    • Maximum of 5 mm
    • Analgesia/Anesthesia is required for mice 21 days of age and older
compound administration
Compound Administration
  • 25 to 27 gauge needle

Maximum Administration Volumes (in ml/kg)

compound administration25
Compound Administration
  • Subcutaneous (SC or SQ)
    • The sides of the mouse serve as good site for SQ injections
    • Insert needle underneath skin
    • Aspirate negative pressure
    • Inject compound and watch for SQ bleb
    • To reduce leakage from the injection site, pause before retracting the needle
compound administration26
Compound Administration
  • Click to Watch Video
compound administration27
Compound Administration
  • Intraperitoneal (IP)
    • Locate lower right quadrant for injection site
    • Aspirate
    • If an unintended subcutaneous bleb occurs, reposition the needle
compound administration28
Compound Administration
  • Click to Watch Video
compound administration29
Compound Administration
  • Intravenous (IV)
    • Lateral tail vein
    • Proper restraint
    • Heat source to dilate vessel
    • Apply direct pressure after injection
compound administration30
Compound Administration
  • Gastric Gavage (PO)
    • Use a bulb-tipped gastric gavage needle
    • Measure length of needle from mouth to last rib
    • DO NOT FORCE the needle down the esophagus
    • Inject solution
    • Observe mouse for signs of distress
compound administration31
Compound Administration
  • Click to Watch Video
  • When planning any procedure involving anesthesia and/or surgery, please consult one of the laboratory animal veterinarians in the Office of Laboratory Animal Care (OLAC) at 974-5634.
  • The veterinarian can provide guidance and detailed information in selecting the most appropriate anesthetic and analgesic protocol for your mice and procedure.
aseptic technique
Aseptic Technique
  • Surgical Prep
    • After induction of anesthesia, clip hair from the surgical site
    • Prep skin with povidone iodine, chlorhexidine or other appropriate skin antiseptic
    • Scrub in a circular pattern, beginning in the center and spiraling outward
    • Follow povidone iodine scrub with a 70% alcohol prep
    • Repeat Twice
    • End procedure with a light coat of povidone iodine solution (not scrub) to the surgical site
aseptic technique34
Aseptic Technique
  • Place a sterile drape over the mouse
  • Anything that touches the surgical site must be sterile
  • Non-absorbable sutures/clips should be removed in 7-14 days
surgical monitoring
Surgical Monitoring
  • Prevent pain, hypoxia, and hypothermia
  • Monitor withdrawal reflex
  • Provide a source of external heat
  • Provide appropriate analgesics for post-operative pain management
  • Inhalant Anesthetic Overdose
    • Isoflurane
    • End the procedure with a thoracotomy or cervical dislocation
  • Carbon Dioxide
    • Place mouse in the chamber
    • Turn on CO2 flow into the chamber
    • Once the mouse has stopped breathing, wait at least 1 minute before removing the mouse from the chamber
    • End the procedure with a thoracotomy or cervical dislocation
    • Substantially prolonged in neonates
  • Cervical Dislocation
    • Performed on anesthetized mice
    • Must be performed by skilled personnel
occupational health and safety
Occupational Health and Safety
  • PPE (Personal Protective Equipment)
    • Protects handler and mouse
    • May include: gloves, gowns, lab coats, shoe covers, hair bonnets, face masks
prevention of infectious disease
Prevention of Infectious Disease
  • Colony Health Surveillance
    • Sentinel mice detect for presence of infectious pathogens and parasites in the research colony
  • Mouse Antibody Production Test (MAP Test) and PCR
    • A test for cell lines and tumors for murine viruses
    • Included in every animal use protocol
procurement of mice
Procurement of Mice
    • APPROVED vendors include Charles River, Jackson Labs, Harlan, NCI-Frederick, and Taconic
    • Mice that are shipped from a non-approved vendor source must be approved by OLAC before the mice are procured.
  • An animal requisition form must be submitted to the facility manager:

  • Quarantine is required if receiving mice from an unapproved vendor
  • The minimum quarantine period is six (6) weeks
  • No experimental manipulations or breeding can be initiated during the quarantine period unless approval has been granted by an OLAC veterinarian
health concerns
Health Concerns
  • General appearance
    • Lethargy
    • Aggressiveness
    • Hunched posture
  • Coat
    • Piloerection
    • Hair loss
    • Unkempt appearance
health concerns44
Health Concerns

Abnormal masses

Skin lesions

health concerns45
Health Concerns
  • Teeth



body condition scoring
Body Condition Scoring

Ullman-Cullere, et al. 1999

tumor production
Tumor Production
  • Four criteria for euthanasia:
    • Tumor size of 1.5 cm in diameter or tumor ulceration
    • Body condition score of 1
    • Tumor interferes with function of vital organs
    • Tumor significantly interferes with locomotion
reporting signs of pain or distress
Reporting Signs of Pain or Distress
  • For Mice That Require Veterinary Care
    • Complete the red “Sick Animal” cage card
    • Attach card to cage
    • Notify facility manager or
      • Print Clinical Case Request Form
      • Fax form to the OLAC office 974-5649
    • Assessment of the mouse’s condition and treatments will be recorded on the “Sick Animal” card
  • Guide for Care and Use of Laboratory Animals.National Academy Press, 2011.
  • Lawson, PT. 2005. Assistant Laboratory Animal Technician (ALAT) Training Manual. The American Association for Laboratory Animal Science, Memphis, TN.
  • Suckow M, Danneman P, Brayton C.  2001. THE LABORATORY MOUSE. CRC Press, Boca Raton, FL.
  • Ullman-Cullere, M.H. and Foltz, C.J. 1999. Body Condition Scoring:  A Rapid and Accurate Method for Assessing Health Status in Mice.  Lab AnimSci 49(3) 319-323.
  • Presentation: Chris Carter, OLAC UTK