Reagents, supplies, and
- #21 surgical blades
- 21 ½ G needle
- insulin syringe
- 5 ml syringe
- human insulin 100 IU
- 0.9 % sterile saline
- BD logistic glucometer
- BD glucose test strips
- Clean mouse cages (with normal bedding: dried corn)
- bench paper
- 250 ml plastic beaker
- 70% EtOH
- Bring mice to lab around 5:30 PM
- Disinfect work area with 10% Clorox, then 70% EtOH
- Prepare work area with balance, clean bench paper, needles,
blades, glucometer, and insulin syringes.
- Allow mice to rest in the lab O/N for ~ 15 hr. Provide
food and water as normal.
- Fast mice for 6 hr between 9:00 AM - 3:00 PM. (Place mice
in a clean cage without food and with H2O).
- Weigh mice at 2:30 PM.
- Calculate the insulin dose for each mouse (based on body
- Prepare 0.25 IU insulin solution by diluting insulin in sterile
saline (and sterile tubes) @ 1:400; mix by vortexing..
- Dispense the required volume of insulin solution for each mouse
into separate 1.5 ml tubes (Volume is calculated as follows: 0.75
IU insulin/kg BW). Vol (μl) = 3 x BW
- Measure blood glucose of the 1st mouse from tail
blood by cutting < 5 mm of the tail tip.
- Perform intraperitoneal injection (i.p.) of insulin. This
is time 0' (3:00 PM). Record blood glucose level(s).
- Continue i.p. injections on subsequent mice, maintaining at
least a 1 min interval between animals.
- Repeat blood glucose measurement at 15', 30', 45', 60', and 90'
by reopening the tail wound each time. There is no need to re-cut
- Upon completion, provide food to each mouse and return the mice
to the mouse colony.
Written by: Yanyun Gu 5/10
itt_worksheet.doc - Added on June 11, 2010 at 11:12 AM by Jill Lindner