Comparison of Cellular Autofluorescence Patterns of Two Model Microalgae by Flow Cytometry

Uğur Uzuner

doi:10.18466/cbayarfbe.827615

Research Article

Comparison of Cellular Autofluorescence Patterns of Two Model Microalgae by Flow Cytometry

Year 2021, , 159 - 165, 28.06.2021

Uğur Uzuner

https://doi.org/10.18466/cbayarfbe.827615

Abstract

Mikroalgler, biyoteknolojik araştırmalarda, özellikle antioksidanlar, sekonder metabolitler, pigmentler, karbohidratlar, proteinler ve lipitler gibi biyokimyasal bileşiklerin üretilmesinde yaygın olarak kullanılmaktadır. Hem deneysel hem de üretim süreçlerinde mikroalglerin değerlendirilmesinde kullanılabilecek çeşitli yöntemlere ihtiyaç duyulmaktadır. Bu yöntemlerden biri olarak akım sitometrisi, mikroalg hücrelerinde nötral ve polar lipit miktarlarının kantifikasyonu ve hücresel morfolojinin saptanmasında kullanılan avantajlı bir seçenektir. Analiz sürecinde etiketleme protokolünü planlamak için hücrelerin otofloresan özelliklerinin hesaba katılması büyük önem taşır. Çünkü otofloresan miktarı, spesifik olarak işaretlenen protein veya lipitlerin floresan sinyali ile çakışabileceğinden, bu moleküllerin tespitini engelleyebilir. Bu durum, hücreler içindeki etiketli bileşiklerin miktarının yanlış anlaşılmasına neden olabilmektedir. Bu çalışmada, endüstriyel önemdeki iki tatlı su mikroalg ümodeli Chlamydomonas reinhardtii (CC-124) ve Chlorella vulgaris’in (CV-898) otofloresan özellikleri akım sitometrisi ölçümleri üzerinden incelenmiştir. Deneysel bulgular, floresan kanal-2'nin, hem (FL2-H) CC-124 hem de CV-898 mikroalg suşlarının minimum otofloresanını elde etmek için en uygun kanal olduğunu göstermiştir. Elde edilen sonuçlar ayrıca florofora karar verirken biyolojik ürünlerin akış sitometrisine dayalı tespiti sırasında CC-124 ve CV-898 hücre hatlarındaki otofloresans sinyallerine dikkat edilmesi gerektiğini ileri sürdü.

Keywords

autofluorescence, cellular morphology, flow cytometry, microalgae

Supporting Institution

Karadeniz Technical University

Project Number

FAY-2016-5755

Thanks

This study was supported by Karadeniz Technical University, with the grant number FAY-2016-5755. The author thanks to Ramazan ÇAKMAK for preparations of cells and to Prof. Ersan KALAY from Flow Cytometry Unit of Department of Medical Biology.

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