Paw Tattooing
Objective: Identification of individual pups through subcutaneous tattooing of the paw with permanent ink at postnatal day 2
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Protocol Steps
Breeding pair setup
Obtain breeding pairs from The Jackson Laboratory and house in standard wire-topped Plexiglas cages in temperature and humidity controlled vivarium with ad libitum food and water access
Note: Lights on from 19:00 to 7:00
View evidence from paper
“BTBR, B6, FVB/NJ, and 129X1 breeding pairs were purchased from The Jackson Laboratory (Bar Harbor, ME). Mice were housed in standard wire-topped Plexiglas cages (42 cm×27 cm×14 cm) in a temperature and humidity controlled vivarium, with food and water available ad libitum, with lights on from 19:00 to 7:00”
Monitor for pregnancy and delivery
Ten days after pairing for breeding, individually house females and inspect daily at 9:30 for pregnancy and delivery. Record day of birth as postnatal day (pnd) 0
Note: The day of birth is designated as pnd 0
View evidence from paper
“Ten days after pairing for breeding, the females were individually housed and subsequently inspected daily at 9:30 for pregnancy and delivery. The day of birth was considered as postnatal day (pnd) 0”
Prepare tattoo ink and needle
Load Ketchum permanent Tattoo Inks green paste into a 30G hypodermic needle
View evidence from paper
“by loading the ink into a 30G hypodermic needle”
Perform paw tattooing
Insert the ink subcutaneously through the needle tip into the center of the paw to create a permanent identification mark
Note: Procedure causes only minor brief pain and distress and does not require anesthesia
View evidence from paper
“inserting the ink subcutaneously through the needle tip into the center of the paw. The procedure causes only minor brief pain and distress and does not require the use of anesthesia”
Timing of tattooing
Perform tattooing procedure at postnatal day 2 (pnd 2), immediately after behavioral testing
Note: Behavioral testing conducted between 9:30 and 14:00 h during dark phase of circadian cycle
View evidence from paper
“The procedure was performed at two days of age, immediately after behavioral testing”