Intra-cardiac Blood Collection in Mice: A Comprehensive Guide

Intra-cardiac Blood Collection in Mice: A Comprehensive Guide

Purpose and Summary of the Procedure

Specifically, it addresses the topic of mice intra-cardiac blood collection methods.
Things to think about

Preparation for the procedure

1. This intra-cardiac blood collection surgery has no chance of survival and is considered a terminal and fatal procedure for mice.
2. The animal must be administered deep general anesthesia to undergo this surgery.
3. To get the proper amounts and lengths of needles, check out the institutional Guidelines for Injections in Rodents and Rabbits.
4. Potentially usable sample volume – 0.5 to 1.0 ml, or 3-5% of total body mass
   Protective Gear of Personal Protective Equipment (PPE) and Hygiene Practices
5. Ensure the technician wears the proper personal protective equipment (PPE) to prevent infection from blood and other bodily fluids. This includes gloves, goggles, a mask, and any other PPE necessary for the procedure or the institution.
6. When handling different animals, washing hands and/or changing gloves is essential.
7. Immediately place used sharps in the supplied container, which is leak-proof and puncture-resistant.
   Chemicals and Supplies
8. Singular or combined administration of a general anesthetics, including but not limited to:
   a. breathed isoflurane—typically about 3%
   b. Inhaled gas mixture of 70% carbon dioxide and 30% oxygen—also breathed to provide the desired effect
   c. A ketamine and xylazine combination of 90-120 mg/kg and 8-12 mg/kg, respectively
9. Injection into a muscle, under the skin (subcutaneous, s.c), or within the abdomen (intraperitoneal)
10. Syringes containing either 1 or 3 cc of tuberculin
11. Needles from 23 to 27 gauge, measuring ⅝-1 inch

Also, see how to prepare single-cell suspension from mice spleen

Frequency of the intra-cardiac blood collection procedure

What is the frequency?

1. The intra-cardiac method is a last resort for the collection of blood and is terminal for mice.
2. Requires anesthesia

Collecting Blood from Mice, Intra-cardiac

Procedure

The necessity of general anesthesia

1. Before collecting samples, thoroughly anesthetize the animal with the anesthetic of your choice.
2. After the animal has achieved the correct level of anesthesia, position it in dorsal recumbency (Figure 1).
3. With the syringe parallel to the mouse’s midline, thread a needle of the correct size into it and enter it through the diaphragm with the bevel facing up at 30–40 degrees (Figure 2).
4. Insert the needle towards the animal’s head, just to the left of the sternum and beneath it. The needle can be inserted at an angle of around 45 degrees toward the left shoulder.
5. After creating a slight vacuum within the syringe, retract the plunger and carefully advance the needle until a blood flash occurs in the needle hub.
6. After gathering enough blood, immobilize the needle and aspirate (Figure 3).
7. Need to euthanize the animal as soon as possible once blood sampling is complete through either a bilateral thoracotomy or a cervical dislocation.

Various types of intra-cardiac blood collection

a. The left-lateral strategy

1. Start by putting the animal in a right-side recumbent position.
2. Feel for the heart on the left side of the chest, between the fifth and sixth ribs, where the elbows are bent.
3. Carefully insert the needle beside the body at a right angle.
   b. Use an open approach
   Figure 1: Appropriate Placement Figure 2. Needle Insertion Angles of 30–40° Blood Aspiration (Figure 3)

The Intra-cardiac Blood Collection in Mice

1. First, get the animal to lie down on its back.
2. Moisten skin with 70% ethanol.
3. Approximately 1 cm below the final rib, make a V-shaped incision into the skin and abdominal wall.
4. Set away abdominal organs.
5. To restart blood circulation, insert or remove the needle through the diaphragm into the vena cava or heart. Spin the needle.

Possible side effects of intra-cardiac procedure, how to reduce them, or treat them

a. Pain from being restrained or from bleeding excessively

1. Execute a supplemental euthanasia procedure
2. Return to the gas anesthesia and ensure continuous administration by wearing the face mask.

See more protocols

https://abdullahfarhan.com/protocol-for-mouse-spleen-isolation-and-preparation-of-single-cell-suspension