Macrophages play unique and critical roles in different parts of the body, however, precise macrophage removal has become a key research tool in order to better study the specific functions of macrophages in specific areas and their impact on disease development. Clodronate Liposomes are one of the most proven, cost-effective and ideal tools for macrophage removal. Clodronate Liposomes can be administered intraperitoneally, in the tail vein, and in various other ways to remove macrophages from different tissues.

In the following, we will share a series of topics to demonstrate the methods and effectiveness of macrophage removal at different sites, hoping to provide useful references and reference for researchers.

Introduction of injection methods

Tail vein injection

Tail vein injection reaches maximum depletion after 24 h. Attention should be paid to the time point of detection.

1、Place the mice in an inverted lunch box, pull out the tail from the orifice, there are two arteries and three veins in the tail of the mice, two arteries are in the dorsal and ventral sides of the tail, and the three veins are distributed in zigzag pattern, generally the left and right sides of the veins are used;

2、Soak the gauze in about 70℃ warm water (or soak it in alcohol), take it out and wrap the tail, in order to achieve the purpose of vasodilatation of the tail and soften the epidermal keratin;

3、For tail intravenous injection, with the left thumb and forefinger before and after holding down the tail of the rat, leaving a section of about 2 cm, so that the skin is tense, the veins are more full, the right hand holding a 1 mL needle syringe, so that the needle and the vein are parallel (less than 30°angle), from the lower 1/4 of the tail into the needle, the beginning of the injection of the drug should be slow, careful observation, if there is no resistance, no white dermatomes appeared, it means that the blood vessels have been pierced and can be Formal injection of drugs.

4、Some experiments require repeated tail vein injection for several days, the injection site should start from the tail end as far as possible, and move to the tail root in sequence, and change the vascular position for injection. After removing the needle, press the injection site with a cotton ball for 1 - 2 min to stop bleeding.

Intraperitoneal injection

Intraperitoneal injection reaches maximum depletion after 48 - 72 h. Attention should be paid to detecting time points.

1、Firstly, use the dorsal bailing method to catch the mice, so that their heads are slightly tilted back and their abdomens are upward, the left hind limb of the mice is fixed with the little finger, and then the injection site is sterilized with a 75% alcohol cotton ball.

2、The location of the intraperitoneal injection in mice is 0.5 cm on both sides of the midline of the lower abdomen (flush with the root of the thigh). In order to avoid injury to the organs, the head of the mouse should be tilted backward when bailing the mouse, so as to make the organs of the lower abdomen move upward.

3、Pierce the syringe needle into the skin, enter the subcutaneous area, push the needle forward along the subcutaneous area for 2 - 3 mm, and then pierce into the abdominal cavity of the mouse at an angle of 45 degrees between the needle and the skin. Note: After penetrating the peritoneum, the resistance of the needle tip disappears.

4、Withdraw the needle plug back, if there is no return of blood or liquid, the drug can be injected.

5、After injection, gently rotate the needle, then slowly pull out the syringe to avoid liquid leakage.

There will also be different injection methods for different study sites, such as: subcutaneous injection, tracheal administration, intracranial injection and so on. You should refer to relevant literature and consult with Yeasen technical team before experimentation.

Dosing cycle and dosage

Short-term administration: a single injection of chlorophosphate liposomes 200 µL (20 - 25 g weight) at a specific time to detect macrophage clearance efficiency or for downstream experiments.

Long-term administration: more than one week or more is required to achieve macrophage clearance. In WT mice, for example, 200 µL (20 - 25 g weight) is usually given as the first dose, followed by 200 µL every 2 - 3 days.

Reference test protocol

Note: The above is for reference only, please refer to relevant literature and consult with Yeasen technical team prior to experimentation.

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