Name: Author Spotlight: Developing an Efficient Blood Collection Method in Non-Anesthetized Rats
Uploaded: 2023-11-03T13:50:00
Description: Watch this Scientific Journal Video about Developing an Efficient Blood Collection Method in Non-Anesthetized Rats at JoVE.com

This study was supported by Cuiying Plan Project of Lanzhou University Second Hospital (Grant No. PR0121015) and Gansu Provincial Key Laboratory of Urinary System Disease Research (Grant No. 0412D2).

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 befor...

Subclavian Vein Puncture Method for Repeated Blood Collection in Conscious Rats

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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 duration4,5. Although high-performance liquid chromatography (HPLC) -tandem mass spectrometry (MS/MS) systems combined with capillary microsampling (CMS) of rat tail ve...

<a href="https://app.jove.com/t/6595">This corrects</a> the article <a href="https://dx.doi.org/10.3791/66075">10.3791/66075</a>

An erratum was issued for: <a href="https://www.jove.com/t/66075">Subclavian Vein Blood Sampling in Conscious Rats</a>. The Discussion section was&#160;updated.

Erratum: Subclavian Vein Blood Sampling in Conscious Rats

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 collection is the most common method for blood sampling from rats, despite not requiring anesthesia, this method can be challenging for repeated collections, and the volume of blood collected is relatively small3,4. Additionally, although blood can be collected from the saphenous and penile veins, the amount of blood obtained is limited, and anesthesia is required1,5. Furthermore, blood samples collected from the submandibular venous plexus, as well as sublingual, jugular, and subclavian veins provide higher-quality samples but typically require anesthesia or the assistance of multiple individuals1,6,7,8,9. Finally, retro-orbital sinus/canal blood collection not only necessitates anesthesia but may also potentially cause injury and stress to the rats9.
The quality of blood samples typically obtained from major veins is generally of the highest standard1. At present, some studies have found that continuous microsampling through the jugular vein is a very suitable method for toxicological research in rats although this method usually requires jugular vein catheterization10,11,12. Therefore, it is worth exploring how to obtain high-quality blood samples in accordance with the 3R principle of animal research without surgical intervention. The objective of this study was to present a method for efficiently extracting blood from the subclavian vein in rats. This technique enables the swift collection of satisfactory samples through a single-person procedure without the need for anesthesia.

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			<td>0.75% normal saline</td>
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			<td>1 mL Pasteur pipette&#160;</td>
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			<td>1 mL syringe (26 G, 0.45 mm x 12 mm)</td>
			<td>Shinva Medical Instrument Co.,Ltd.</td>
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			<td>1.5 mL Eppendorf tube</td>
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			<td>75% medical alcohol</td>
			<td>Shandong Lircon Medical Technology Co., Ltd.</td>
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			<td>Disinfection of rat blood collection site</td>
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			<td>Centrifuge tube holder</td>
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			<td>Electronic scale</td>
			<td>Shanghai PUCHUN Measure Instrument Co., Ltd.</td>
			<td>JE1002</td>
			<td>Weigh</td>
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			<td>Heparin sodium injection</td>
			<td>Hebei Changshan Biochemical Pharmaceutical Co., Ltd.</td>
			<td>&#8212;&#8212;</td>
			<td>Rinse the syringe and EP tube; dilute with normal saline to 25 U/mL</td>
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			<td>Low temperature centrifuge</td>
			<td>HuNan Xiang Yi Centrifuge Instrument&#160; Co., Ltd.</td>
			<td>&#160;H1750R</td>
			<td>Separation of serum</td>
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subclavian vein blood sampling in conscious rats

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.

Author Spotlight: Developing an Efficient Blood Collection Method in Non-Anesthetized Rats 

Subclavian Vein Blood Sampling in Conscious Rats

The scope of our research is centered on devising infection methodologies for collecting blood samples from rats. Our aim is to ensure rapid and set procedures that align with our principle, utilizing minimal manpower and resources. One of the current experimental challenges is to effectively immobilize the rat in a manner that simultaneously maintains the subclavian vein enclosed proximately to the skin surface for optimal blood collection.The advantages of our protocol is that high quality blood samples can be obtained quickly by a person without anesthesizing the rats.

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 &#177; 33.62 s and 123.28 &#177; ...

Watch this Scientific Journal Video about Developing an Efficient Blood Collection Method in Non-Anesthetized Rats at JoVE.com

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.

There are several established methods for obtaining repeated blood samples from rats, with the most commonly employed methods being lateral tail vein ...