Laboratory Manual
Course name: Pharmacology (2) Lab
Course number: 304337
Lab #1: Introduction and Orientation
This is a pharmacology lab in which responses to drugs are observed in
animals. In animal studies various types of animals might be used including
rats, rabbits, monkeys, and mice. Small animals such as mice and rats are
preferred in most pharmacologic research and for teaching purposes,
especially when a large number of candidates are required. The type of
animals used is determined based experiment’s objectives; for example, in
experiments focusing on organ/tissue studies such as studying the
hepatoxicity of a certain drug, rats are preferred over mice due to their larger
size. While in other experiments where the focus is not on organs, like in
screening for efficacy or screening for toxicity of certain drug mice are
preferred.
In this course (pharmacology lab) Balb-c mice are used as the animal model
(Figure 1). The vast majority of the conducted experiments are quantitative
i.e. the effects of specific drugs are measured and compared to the controls.
Figure 1 Balb- C mouse
LAB #2: Mouse Handling and Injection
- Objective:
The objective of this lab session is to learn the proper mouse handling and
intraperitoneal injection method.
- Introduction
Handling and restraining are the most important techniques for correct
administration of a test medication/substance. Appropriate restraining leads
to successful administration, and the restraining techniques vary based on
the route of administration. In this lab “Double handed manual restraint” is
the restraining technique used, Figure 2.
Figure 2. Manual restraint of a mouse using both hands. (a) The mouse is placed on the cage lid. The tail is pulled gently back.
(b) The mouse is quickly and firmly picked up by the scruff of the neck behind the ears with thumb and index finger of other
hand. (c) The tail is transferred from the preferred hand to between palm and little or ring finger of the other hand, then
fixed. (d) The mouse is restrained.
Several routes of administration are used for the administration of a
medication/substance to mice, including:
a) Oral gavage that involves inserting the feeding needle through mouth into
the esophagus and is usually used when direct dosing is required.
b) Parenteral administration, including:
1. intravenous administration (IV) where the substance/medication is
injected in the tail, it is very hard and requires very skilled personnel.
2. Subcutaneous administration (SC) where the substance is injected in
the scruff area of the neck (the area between the two ears).
3. Intraperitoneal administration (IP) that represent the most common
route for injection in animal models, it is a simple and easy route for
injection, It offers fast rate, and extinction of absorption, it involves
injecting in the lower quadrant of the abdomen.
Dose administration is performed using insulin syringe which is labeled from 10-
100 international units in which each 100-international unit = 1 ml. The insulin
syringe could have varying length and gauge values. Gauge value represent needle
thickness, and as the gauge value increases thickness decreases. Lastly, the
maximum volume that could be injected to a mouse is 2 ml (2000 µl).
- Steps:
1. Prepare an injection of 100 ul of saline using the insulin syringe
2. Gently take out the Balb-c mouse from its cage
3. Put it gently on the steel grid holding it from the middle of the tail
4. Place your other hand on its scruf, turn the mouse to your side and place
the tail between your fingers
5. Incline the syringe at 45° and inject the dose in the upper left abdominal
quadrant
6. Repeat this procedure
LAB#3: Dose calculation and administration to the mouse
- Objective:
The objective of this lab session is to calculate and inject a proper dose to a
given mouse.
- Method:
A theoretical example will be used here. Suppose you were given a mouse of
25g, a stock solution of 75mg/3ml of drug “X” and you need to administer a
dose of 100mg/kg. What would be the volume of the solution needed to be
injected into the mouse via the I.P route?
Steps:
1. Take the mouse from the cage
2. Mark the mouse on its tail
3. Weigh the mouse
4. Calculate the volume needed as mentioned previously
5. Inject the proper dose as in the previous lab
