- Journal of Istanbul Veterinary Sciences
- Vol: 3 Issue: 3
- Polymorphonuclear leukocyte isolation from venous blood of the dog
Polymorphonuclear leukocyte isolation from venous blood of the dog
Authors : Güneş Karakurt, Kader Yildiz
Pages : 75-78
Doi:10.30704/http-www-jivs-net.621263
View : 10 | Download : 2
Publication Date : 2019-12-31
Article Type : Research
Abstract :The current study aimed to isolate neutrophils from venous blood samples of healthy dogs. Venous blood samples were obtained from Venae cephalica of clinically healthy dogs (n:5) into heparinized tubes. The blood samples (2 mL) and Percoll dilutions (45%, 54%, 63%, and 72%) prepared with Hanks Balanced Salt Solution were layered into sterile tubes. After centrifuge, the polymorphonuclear leukocytes (PMN) were aspirated between 63% and 72% interfaces of the Percoll dilutions into tubes. The samples of PMN observed under a light microscope. Viability was detected microscopically after stained with trypan blue dye. Diff-Quick staining was used to detect neutrophil purity of the isolated PMN. In the present study, the neutrophils ratio was calculated as 92% of the isolated polymorphonuclear cells. The neutrophil viability was calculated as 98% of PMNs isolated from the venous blood samples of healthy dogs. In the present study, the Percoll gradient centrifugation (72%, 63%, 54%, and 45%) is a fast technic for isolation of the neutrophils from venous blood samples of dogs.Keywords : dog, venous blood, polymorph nuclear leucocyte, neutrophil, isolation