Subclavian Vein Blood Sampling in Conscious Rats

Summary

Here we present a combination of effective rat restriction and subclavian vein puncture methods that enable rapid, safe, and repeated blood collection in rats without anesthesia.

Abstract

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein sampling without anesthesia and jugular vein sampling with anesthesia. However, most of these methods require assistance and anesthetic equipment and sometimes pose difficulties in terms of blood collection or the poor quality of blood samples. In addition, these methods of blood collection consume significant time and human resources when repeated blood sampling is required for a large number of rats. This study presents a technique for repetitive blood sampling in non-anesthetized rats by a single proficient individual. Highly satisfactory blood samples can be obtained by puncturing the subclavian vein. The method demonstrated an impressive overall success rate of 95%, with a median time of merely 2 min from rat restraint to the completion of blood collection. Furthermore, performing consecutive blood collections within the designated range does not inflict any harm on the rats. This method is worth promoting for blood collection, especially in large-scale pharmacokinetic studies.

Introduction

Rats are one of the most common experimental animals, and there are many ways to obtain blood samples. For experiments involving a single blood collection at the concluding stage, a sufficient amount of blood can be obtained through cardiac puncture or abdominal aorta blood collection¹. However, some studies require repeated blood collection from rats for routine blood or biochemical analysis, especially in pharmacokinetics and toxicology studies, where repeated blood collection is required to determine the absorption, distribution, and metabolism of drugs².

Currently, although tail vein blood....

Protocol

This study adhered to the guidelines outlined in the 8th edition of the Guide for the Care and Use of Laboratory Animals¹³. The research received approval from the Ethics Committee of Lanzhou University Second Hospital and was documented in adherence to The ARRIVE guidelines 2.0¹⁴. Twelve healthy Wistar rats (six males weighing 290-330 g and six females weighing 250-280 g) aged 12-16 weeks were accommodated in the GLP Animal Laboratory of Lanzhou University for 3 days before the actual experiment. The rat cages utilized were of the R5 type, measuring 545 mm x 395 mm x 200 mm, and were equipped with autoclaved bedding mat....

Results

High-quality plasma specimens exhibit a pale-yellow hue, clarity, and transparency, devoid of any red tinge or clotting, as depicted in Figure 2A. Figure 2B shows hemolysis (left side) or coagulation (right side) as a result of improper procedures, respectively. Over the course of 96 blood collection sessions within 4 days, the average single blood collection times for groups A and B were 119.87 ± 33.62 s and 123.28 ± 30.96 s, r.......

Discussion

Although tail vein blood collection is the most common method for repeated blood sampling in rats, it might be influenced by anesthesia drugs, and due to the small size of the tail vein, the amount of blood that can be collected in a single instance is limited, leading to a longer blood collection duration^4,5. Although high-performance liquid chromatography (HPLC) -tandem mass spectrometry (MS/MS) systems combined with capillary microsampling (CMS) of rat tail ve.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.75% normal saline	Gansu Fuzheng Pharmaceutical Technology Co., Ltd.	——	Prepared heparin sodium solution
1 mL Pasteur pipette	Biosharp	BS-XG-01-NS	Blood collection
1 mL syringe (26 G, 0.45 mm x 12 mm)	Shinva Medical Instrument Co.,Ltd.	0.45*12RWLB	Blood collection
1.5 mL Eppendorf tube	Biosharp	BS-15-M	Blood storage and collection
75% medical alcohol	Shandong Lircon Medical Technology Co., Ltd.	——	Disinfection of rat blood collection site
Centrifuge tube holder	Biosharp	BS-05/15-SM60	——
Electronic scale	Shanghai PUCHUN Measure Instrument Co., Ltd.	JE1002	Weigh
Heparin sodium injection	Hebei Changshan Biochemical Pharmaceutical Co., Ltd.	——	Rinse the syringe and EP tube; dilute with normal saline to 25 U/mL
Low temperature centrifuge	HuNan Xiang Yi Centrifuge Instrument  Co., Ltd.	H1750R	Separation of serum