JoVE Logo
Faculty Resource Center

Sign In

Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Acknowledgements

Materials

References

Medicine

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Published: June 30th, 2023

DOI:

10.3791/65513

1Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, 2Harvard Medical School, 3Shriners Children’s Boston, 4Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, 5Department of Vascular Surgery, Centre Hospitalier Universitaire Vaudois and University of Lausanne, 6Department of Plastic, Reconstructive and Hand Surgery, University Medical Center Utrecht, 7Department of Plastic, Reconstructive and Aesthetic Surgery, Rennes University Hospital Center (CHU de Rennes), Rennes 1 University, 8Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, 9Plastic Surgery Research Laboratory, Massachusetts General Hospital
* These authors contributed equally

Here, we present a protocol to offer rapid, easy, and reliable blood collection alternatives for the rat model. We describe three different blood sampling methods according to the context: tail vein puncture under anesthesia or on a conscious animal, and dorsal penile vein puncture under anesthesia.

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood collection in rats: the lateral tail vein puncture and the dorsal penile vein puncture, which offer significant advantages over other previously described techniques. This study shows that these two procedures allow for fast sampling (under 10 min) and yield sufficient blood volumes for most assays (202 μL ± 67.7 μL). The dorsal penile vein puncture must be done under anesthesia, whereas the lateral tail vein puncture can be done on a conscious, restrained animal.

Alternating these two techniques, therefore, enables blood draw in any situation. While it is always recommended for an operator to be assisted during a procedure to ensure animal welfare, these techniques require only a single operator, unlike most blood sampling methods that require two. Moreover, whereas these previously described methods (e.g., jugular stick, subclavian vein blood draw) require extensive prior training to avoid harm to or death of the animal, tail vein and dorsal penile vein puncture are rarely fatal. For all these reasons, and according to the context (e.g., for studies including male rats, during the perioperative or immediate postoperative period, for animals with thin tail veins), both techniques can be used alternately to enable repeated blood draws.

Blood sampling is necessary for most animal studies, both in vivo and in vitro. In rats, as the frequency and amount of blood sampling can be significant, it is helpful to have different alternatives for collection. Various methods have been described in previous studies.

The most commonly used techniques are tail vein puncture and saphenous vein blood draw. Tail vein sampling is suitable for all rat strains. With proper training, the procedure is simple to perform and causes minimal distress to the animal1. Similarly, the saphenous vein blood draw, provided it is done properly, is also a quick and ....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

The procedures were performed on 3 month old male Lewis rats, each weighing 300-400 g. A total of 24 animals were included, with three puncture conditions: 12 rats underwent tail vein puncture without anesthesia (group TV without anesthesia), and another 12 rats were anesthetized to undergo both tail vein puncture (group TV with anesthesia) and penile vein puncture (group PV with anesthesia). All the procedures were approved and respected the Institutional Animal Care and Use Committee (IACUC) guidelines. All the animals.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Success was defined as a blood draw yielding at least 100 µL of blood in under 10 min (from the puncture time to the end of the blood collection), and failure was defined as a blood draw yielding less than 100 µL of blood or taking more than 10 min to retrieve the required blood volume. A maximum of 250 µL of blood per sample was allowed. The statistical analyses were conducted using a one-way ANOVA test for multiple comparisons and the chi-squared test. The data were presented as mean value ± standar.......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

The tail vein puncture is an efficient method to obtain blood from a conscious rat. However, when an animal is under anesthesia, the effect of isoflurane can lead to vessel spasms and make tail vein puncture unsuitable11. As shown in this study, an alternative in this situation is to collect blood from the penile vein, which is more successful and yields a significantly greater volume of blood in less time. It is important to remember that in the case of failure with this method on the first try, .......

Log in or to access full content. Learn more about your institution’s access to JoVE content here

This work was funded by Shriners Children's Boston (B. E. U., K.U., C.L.C.). L.C. is funded by "La Bourse des Gueules Cassées", "La Bourse Année Recherche", and "La Bourse de l'Amicale des Anciens Internes des Hôpitaux de Paris". Y.B. is funded by "La Bourse des Gueules Cassées". Y.B. and I.F.v.R. are funded by the Shriners Hospitals for Children (Fellowships ID is #84308-BOS-22 #84302-BOS-21 respectively). This material is partially based upon work supported by the National Science Foundation under Grant No. EEC 1941543. Partial support from the US National Institutes of Health (R01EB028782, R56AI171958, and....

Log in or to access full content. Learn more about your institution’s access to JoVE content here

Name Company Catalog Number Comments
0.5 mL | 28 G ½  Insulin Syringes BD 329424 for tail vein puncture
0.5 mL | 30 G x 5/16 Insulin Syringes BD 320468 for penile vein puncture
250 L  Microtainer blood collection tubes with K2EDTA BD 365974
Gauze Sponges Curity 6939
Isoflurane Auto-Flow Anesthesia Machine E-Z Systems EZ-190F for penile vein puncture
Isoflurane, USP Patterson Veterinary 1403-704-06 for penile vein puncture
Nosecone for Anesthesia World Precision Instruments EZ-112 for penile vein puncture
Rodent Restraint Cone Harvard Apparatus ST2 52-95-86 for tail vein puncture
Small Animal Heated Operating Table (Adjustable) Peco Services Ltd 69023
Webcol Alcohol prep pads Simply Medical 5110

  1. Lee, G., Goosens, K. A. Sampling blood from the lateral tail vein of the rat. Journal of Visualized Experiments. (99), e52766 (2015).
  2. Beeton, C., Garcia, A., Chandy, K. G. Drawing blood from rats through the saphenous vein and by cardiac puncture. Journal of Visualized Experiments. (7), 266 (2007).
  3. Luzzi, M., et al. Collecting blood from rodents: A discussion by the Laboratory Animal Refinement and Enrichment Forum. Animal Technology and Welfare. 4 (2), 99-102 (2005).
  4. Wang, L., et al. Repetitive blood sampling from the subclavian vein of conscious rat. Journal of Visualized Experiments. (180), e63439 (2022).
  5. Van Herck, H., et al. Blood sampling from the retro-orbital plexus, the saphenous vein and the tail vein in rats: Comparative effects on selected behavioural and blood variables. Laboratory Animals. 35 (2), 131-139 (2001).
  6. Harikrishnan, V. S., Hansen, A. K., Abelson, K. S., Sørensen, D. B. A comparison of various methods of blood sampling in mice and rats: Effects on animal welfare. Laboratory Animals. 52 (3), 253-264 (2018).
  7. Zeller, W., Weber, H., Panoussis, B., Bürge, T., Bergmann, R. Refinement of blood sampling from the sublingual vein of rats. Laboratory Animals. 32 (4), 369-376 (1998).
  8. Nightingale, C. H., Mouravieff, M. Reliable and simple method of intravenous injection into the laboratory rat. Journal of Pharmaceutical Sciences. 62 (5), 860-861 (1973).
  9. Blood collection: The rat. IACUC Guideline. UCSF Office of Research Institutional Animal Care and Use Program Available from: https://iacuc.ucsf.edu/sites/g/files/tkssra751/f/wysiwyg/guidelines%20-%20Blood%20Collection%20-%20Rat.pdf (2022)
  10. Staszyk, C., Bohnet, W., Gasse, H., Hackbarth, H. Blood vessels of the rat tail: A histological re-examination with respect to blood vessel puncture methods. Laboratory Animals. 37 (2), 121-125 (2003).
  11. Constantinides, C., Mean, R., Janssen, B. J. Effects of isoflurane anesthesia on the cardiovascular function of the C57BL/6 mouse. ILAR Journal. 52 (3), e21-e31 (2011).
  12. Hernaningsih, Y., Akualing, J. S. The effects of hemolysis on plasma prothrombin time and activated partial thromboplastin time tests using photo-optical method. Medicine. 96 (38), 7976 (2017).
  13. Powles-Glover, N., Kirk, S., Jardine, L., Clubb, S., Stewart, J. Assessment of haematological and clinical pathology effects of blood microsampling in suckling and weaned juvenile rats. Regulatory Toxicology and Pharmacology. 69 (3), 425-433 (2014).

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved