A new approach for classification of major larval hemocytes (prohemocytes, plasmatocytes and granulocytes) in the greater wax moth, Galleria mellonella L. (Lepidoptera: Pyralidae) by acridine orange staining

Abstract

Larval hemocytes of the greater wax moth, Galleria mellonella L. 1758 (Lepidoptera: Pyralidae), were classified by fluorescence microscopy staining with acridine orange. Based on their fluorescence characteristics, plasmatocytes and prohemocytes could be easily distinguished from granulocytes because they appear faint green and do not contain any fluorescent granules in their cytoplasm. Nuclei of granulocytes emit bright green fluorescence when observed under fluorescence microscope. Prohemocytes are round and their nuclei are bright. Plasmatocytes are irregularly shaped. Therefore, all three types of the wax moth’s major hemocytes that are difficult to identify by bright field or phase contrast microscopy can be easily classified by fluorescence microscopy with staining acridine orange. Furthermore, this technique will assist further studies to understand how hemocyte differentiation and regeneration proceeds in larval hematopoietic organs and during hemocyte transformation. The fluorescent method for hemocyte classification is more precise than the common method of hemocyte identification using bright field or phase contrast microscopy.

Keywords

• Galleria mellonella
• hemocyte
• acridine orange
• fluorescence microscopy

Büyük bal mumu güvesi Galleria mellonella L. (Lepidoptera: Pyralidae)'nın başlıca larval hemositlerinin (prohemositler, plazmatositler ve granülositler) akridin oranj boyama ile sınıflandırılması için yeni bir yaklaşım

Öz

Büyük mum güvesi, Galleria mellonella L. 1758 (Lepidoptera: Pyralidae)’nın larval hemositleri acridine orange ile boyanarak floresans mikroskopta sınıflandırılmıştır. Hemositlerin floresan özelliklerine göre, plasmatositler ve prohemositler, granulositlerden kolaylıkla ayırt edilebilir. Çünkü plasmatositler ve prohemositler soluk yeşil görünürler ve sitoplazmalarında herhangi bir floresan granülleri içermezler. Granulositler floresans mikroskop ile incelendiğinde nukleusları parlak yeşil ışıma yaparlar. Prohemositler yuvarlaktır ve nukleusları parlaktır. Plasmatositlerin düzensiz şekilleri vardır. Büyük mum güvesinin en önemli üç tip hemositlerin parlak alan ve faz kontrast mikroskop ile tanımlanmaları zor olduğundan dolayı, hemositler acridine orange boyası ile floresans mikroskop altında sınıflandırılabilir. Ayrıca, bu teknik hemositlerin dönüşümü sırasında ve hematopoietik organda hemositlerin nasıl farklılaştığının anlaşılması için yapılan ileri çalışmalara destek sağlayacaktır. Hemositlerin sınıflandırılmasında kullanılan floresan tekniği, genel bir yöntem olan parlak alan ve faz kontrast mikroskop ile yapılan hemosit belirlenmesine göre daha hassas bir yöntemdir

Anahtar Kelimeler

• Galleria mellonella
• hemosit
• akridin oranj
• floresan mikroskobu

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