Introducing a combined Leishman-Giemsa stain as a new staining technique for avian blood smears

Document Type: Short paper

Authors

1 BSc Student in Veterinary Laboratory Science, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran

Abstract

Background: The use of conventional stains including Giemsa, Wright and Leishman have become an essential tool for differential diagnosis of cells in peripheral blood. Aims: The aim of the study was to develop a new combination of Leishman-Giemsa (L&G) stain for avian blood smears and to compare its efficacy with conventional staining methods. Methods: Three sets of peripheral blood smears, one smear for L&G stain and two other smears for Leishman and Giemsa stains, created from 50 broiler chickens blood samples. All the three sets of slides were blind screened by two expert clinical pathologists and scored based on the staining characteristics (4 parameters) such as nuclear features of red blood cell (RBC) and white blood cell (WBC), cytoplasmic features and cytoplasmic granularity of WBC. The average grading score assigned by two experts for each staining method were compared. Results: The average grading score of two conventional Leishman and Giemsa staining methods were significantly lower (P<0.001) than new L&G staining method in avian nuclear features of the RBC and WBC. The L&G stain gave a better clarity of nuclear features of avian RBC and WBC. The new L&G staining technique created significant differences (P<0.001) in cytoplasmic features of avian WBC compared to the other two methods. Conclusions: For the first time, the results of the present study showed that the avian blood cells are more desirable stained with a new combination of L&G stain. In addition, it gives a better nuclear and cytoplasmic differential staining than the conventional Giemsa and Leishman stains when used alone.

Keywords


Belgaumi, UI and Shetty, P (2013). LeishmanGiemsa cocktail as a new, potentially useful cytological technique comparable to Papanicolaou staining for oral cancer diagnosis. J. Cytol., 1: 18-22.

Doddagowda, SM; Shashidhar, HA and Prasad, CS (2017). Leishman-Giemsa cocktail-is it an effective stain for air dried cytology smears. J. Clin. Diagn. Res., 3: 16-18.

Gajendra, S; Jha, B; Goel, S; Sahni, T; Sharma, R; Shariq, M; Jaiswal, S and Sachdev, R (2015). Leishman and Giemsa stain: a new reliable staining technique for blood/bone marrow smears. Int. J. Lab. Hematol., 6: 774-782.

Garbyal, RS; Agarwal, N and Kumar, P (2006). Leishman-Giemsa cocktail. Acta Cytol., 4: 403-406.

Kass, L; Harrison, GJ and Lindheimer, C (2002). A new stain for identification of avian leukocytes. Biotech. Histochem., 4: 201-206.

Kondo, M; Yasumoto, S and Takahashi, Y (2011). Morphological and cytochemical characteristics of leukocytes from Redlip Mullet, Chelon haematocheilus. J. Natl. Fisheries Univ. 59: 173-178.

Kondo, M; Yasumoto, S and Takahashi, Y (2012). Staining characteristics of lymphocyte, monocyte and eosinophil from Japanese Lates, Lates japonicus. J. Natl. Fisheries Univ. 60: 95-102.

Nakada, K; Fujisawa, K; Horiuchi, H and Furusawa, S (2014). Studies on morphology and cytochemistry in blood cells of ayu Plecoglossus altivelis altivelis. J. Vet. Med. Sci., 76: 693-704.

Robertson, GW and Maxwell, MH (1990). Modified staining techniques for avian blood cells. Br. Poult., Sci., 4: 881-886.

Suryalakshmi, S; Thangamani, P; Ravi, S; Niveditha, T; Kumar, IVS; Premalatha, DR; Sasikala, DR and Mohan, DR (2016). Comparision of leishman staining with H&E staining technique in the study of FNAC smears. Int. Organ. Sci. Res., 15: 34-39.

Teerasaksilp, S; Wiwanitkit, V and Lekngam, P (2005). Comparative study of blood cell staining with wright-giemsa stain, field stain, and a new modified stain. Int. J. Lab. Hematol., 1: 76-78.