Laboratory Animals > Volume 43, Number 1 > Pp. 65-71

This Article Figures Only Full Text Full Text (PDF) All Versions of this Article: la.2008.007075v1 43/1/65    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Christensen, S D Articles by Hansen, A K Search for Related Content PubMed PubMed Citation Social Bookmarking                      What's this?

Papers

Quality of plasma sampled by different methods for multiple blood sampling in mice

S D Christensen ¹, L F Mikkelsen ², J J Fels ², T B Bodvarsdóttir ² and A K Hansen ¹ 

¹ Centre for Applied Laboratory Animal Research (www.calar.dk), Department of Veterinary Pathobiology, Faculty of Life Sciences, University of Copenhagen, Dyrlaegevej 88, DK-1870 Frederiksberg C, Denmark; ² Centre for Applied Laboratory Animal Research (www.calar.dk), Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark

Corresponding author: Professor Axel Hansen. Email: akh{at}life.ku.dk

For oral glucose tolerance test (OGTT) in mice, multiple blood samples need to be taken within a few hours from conscious mice. Today, a number of essential parameters may be analysed on very small amounts of plasma, thus reducing the number of animals to be used. It is, however, crucial to obtain high-quality plasma or serum in order to avoid increased data variation and thereby increased group sizes. The aim of this study was to find the most valid and reproducible method for withdrawal of blood samples when performing OGTT. Four methods, i.e. amputation of the tail tip, lateral tail incision, puncture of the tail tip and periorbital puncture, were selected for testing at 21°C and 30°C after a pilot study. For each method, four blood samples were drawn from C57BL/6 mice at 30 min intervals. The presence of clots was registered, haemolysis was monitored spectrophotometrically at 430 nm, and it was noted whether it was possible to achieve 30–50 µL blood. Furthermore, a small amount of extra blood was sampled before and after the four samplings for testing of whether the sampling induced a blood glucose change over the 90 min test period. All methods resulted in acceptable amounts of plasma. Clots were observed in a sparse number of samples with no significant differences between the methods. Periorbital puncture did not lead to any haemolysed samples at all, and lateral tail incision resulted in only a few haemolysed samples, while puncture or amputation of the tail tip induced haemolysis in a significant number of samples. All methods, except for puncture of the tail tip, influenced blood glucose. Periorbital puncture resulted in a dramatic increase in blood glucose of up to 3.5 mmol/L indicating that it is stressful. Although lateral tail incision also had some impact on blood glucose, it seems to be the method of choice for OGTT, as it is likely to produce a clot-free non-haemolysed sample, while periorbital sampling, although producing a high quality of sample, induces such a dramatic change in blood glucose that it should not be applied for OGTT in mice.

Key Words: Mice • blood sampling • oral glucose tolerance test • blood glucose

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?