黑水虻幼虫血细胞类型的研究

Study on hemocytes classification of Hermetia illucens ( L. ) larve

黑水虻Hermetia illucens(L.)是一种重要的资源昆虫。本文旨在筛选出适合黑水虻血细胞观察的染色方法,明确黑水虻血细胞类型、数量及组成,为黑水虻血细胞免疫研究奠定基础。采用Giemsa和Giemsa-Wright's染色方法和血球计数板法,对黑水虻血细胞染色方法和血细胞数量及形态进行研究。结果表明,甲醇固定4 min,Giemsa-Wright's染液染色9 min、pH 7.2磷酸盐缓冲液分色10 min是黑水虻幼虫血细胞最佳染色方法;黑水虻幼虫血细胞包括原血细胞、浆血细胞、粒血细胞、类绛血细胞、珠血细胞5类;4龄黑水虻幼虫血细胞数量大约为2917个/μL,其中浆血细胞占53.20%±2.78%,粒血细胞占37.49%±3.96%,原血细胞占7.97%±1.51%,类绛血细胞占1.02%±0.24%,珠血细胞占0.62%±0.08%。Giemsa-Wright's染色法为黑水虻幼虫血细胞最佳染色方法,黑水虻幼虫血细胞可分为5类10种。

Hermetia illucens（ L.） is a kind of important resource insect. In order to lay the foundation for the research of Haemocyte immune functions of H. illucens,the type,number and composition of the hemocytes of the 4^th stage larva of the H. illucens were studied by selecting suitable staining method which is beneficial to observe the hemocytes of 4^th stage larva of the H. illucens. Hemocytes＇ s quantity and morphology of the 4^th stage larva of the H. illucens was researched by using Giemsa and Giemsa-Wright＇s staining method and blood count plate. Fixing 4 min with Methanol,staining 9 min with Giemsa-Wright＇s,separating color 10 min with pH 7. 2 phosphate buffer is the best method. The hemocytes of the 4^th stage larva of H. illucens could be divided into five types： prohemocyte,plasmatocyte,granulocyte,sphrulocyte and oenocytoid. The number of total hemocytes of the 4^th stage larva of H. illucens was 2917/μL,and the percentage of prohemocyte,plasmatocyte,granulocyte,sphrulocyte and oenocytoid were about 7. 97% ±1. 51%,53. 20% ± 2. 78%,37. 49% ± 3. 96%,0. 62% ± 0. 08%,1. 02% ± 0. 24%,respectively.Giemsa-Wright ＇s method is the suitable method for hemocytes identification. The hemocytes of the 4^th stage larva of H. illucens can be divided into five main types with ten subtypes.