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New approaches in tail‐bleeding assay in mice: improving an important method for designing new anti‐thrombotic agents
Author(s) -
Saito Max Seidy,
Lourenço André Luiz,
Kang Hye Chung,
Rodrigues Carlos Rangel,
Cabral Lucio Mendes,
Castro Helena Carla,
Satlher Plínio Cunha
Publication year - 2016
Publication title -
international journal of experimental pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.671
H-Index - 72
eISSN - 1365-2613
pISSN - 0959-9673
DOI - 10.1111/iep.12182
Subject(s) - repeatability , reagent , lysis , reproducibility , saline , chromatography , chemistry , medicine , immunology
Summary This report describes a modified, simple, low‐cost and more sensitive method to determine bleeding patterns and haemoglobin concentration in a tail‐bleeding assay using BALB /c mice and tail tip amputation. The cut tail was immersed in Drabkin's reagent to promote erythrocyte lysis and haemoglobin release, which was monitored over 30 min. The operator was blinded to individual conditions of the mice, which were treated with either saline (NaCl 0.15 m ), DMSO (0.5%) or clinical anti‐thrombotic drugs. Our experimental protocols showed good reproducibility and repeatability of results when using Drabkin's reagent than water. Thus, the use of Drabkin's reagent offered a simple and low‐cost method to observe and quantify the bleeding and rebleeding episodes. We also observed the bleeding pattern and total haemoglobin loss using untreated animals or those under anti‐coagulant therapy in order to validate the new Drabkin method and thus confirm that it is a useful protocol to quantify haemoglobin concentrations in tail‐bleeding assay. This modified method provided a more accurate results for bleeding patterns in mice and for identifying new anti‐thrombotic drugs.