Sains Malaysiana 49(12)(2020): 2901-2912

http://dx.doi.org/10.17576/jsm-2020-4912-03

Rapid Manipulation of Extracellular Vesicles using Dielectrophoretic Mechanism

(Manipulasi Pantas Vesikel Ekstrasel menggunakan Mekanisme Dwielektroforesis)

NUR MAS AYU JAMALUDIN¹, MUHAMMAD KHAIRULANWAR ABDUL RAHIM¹, AZRUL AZLAN HAMZAH¹, NADIAH ABU² & MUHAMAD RAMDZAN BUYONG¹*

¹Institute of Microengineering and Nanotechnology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²UKM Medical Molecular Biology Institute (UMBI), UKM Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Diserahkan: 4 Mei 2020/Diterima: 23 Jun 2020

ABSTRACT

Extracellular vesicles (EVs) are small entities that are released by most cell types. EVs are important form of intercellular communication and a rich source of biomarkers for a wide variety of diseases. Many methods for EVs isolation have been utilized, however, most of them have significant drawbacks including lengthy processing time, high cost, shortfalls in selectivity and surface marker dependency. In consideration of these issues, this paper discussed on the dielectrophoresis (DEP) microelectrode method designed to rapidly isolate EVs from its medium. The advantage of this DEP microelectrode is the capability of isolating EVs using a droplet of 1 µL placed onto the microelectrode within 30 s and 20 V peak-to-peak (Vp-p) of alternating current (AC). The method used in the characterization of sample are dynamic light scattering (DLS) and transmission electron microscopy (TEM); both prove the heterogeneity of EVs’ population and the EVs appear to be spherical with size ranging from 40 to 200 nm. The experimental results from this preliminary experiment show that the DEP microelectrode was able to manipulate EVs as evidenced by the negative dielectrophoresis (NDEP) fluorescent images. Further investigation of grid analysis conducted shows the consistency of the theory and the results presented. Corrected Total Cell Fluorescence (CTCF) values from the grid analysis concluded that the EVs were manipulated to the center of region of interest (ROI). Therefore, this DEP technique suggests a rapid strategy for EVs isolation from its medium in small quantity while maintaining accuracy and cost-effectivity.

Keywords: Dielectrophoresis; extracellular vesicles; isolation; manipulation; particle separation

ABSTRAK

Vesikel ekstrasel (EV) adalah entiti kecil yang dilepaskan oleh kebanyakan jenis sel. EV adalah bentuk komunikasi antara sel yang penting dan merupakan sumber biopenanda yang kaya untuk pelbagai jenis penyakit. Banyak kaedah untuk pengasingan EV telah digunakan, namun, kebanyakan daripada mereka mempunyai kekurangan yang signifikan termasuk waktu pemprosesan yang panjang, kos yang tinggi, kekurangan pilihan dan kebergantungan penanda permukaan. Berdasarkan permasalahan ini, kajian ini membincangkan kaedah dielektroforesis (DEP) mikroelektrod yang dirancang untuk mengasingkan EV dengan cepat daripada mediumnya. Kelebihan mikroelektrod DEP ini adalah kemampuannya mengasingkan EV menggunakan titisan 1 µL yang diletakkan ke atas mikroelektrod dalam masa 30 saat dan voltan arus ulang alik (AC) yang menggunakan 20 volt puncak-ke-puncak (Vp-p). Kaedah yang digunakan dalam pencirian sampel adalah penyerakan cahaya dinamik (DLS) dan mikroskop elektron transmisi (TEM); kedua-duanya membuktikan keheterogenan populasi EV dan EV kelihatan bulat dengan ukuran antara 40 nm hingga 200 nm. Hasil daripada uji kaji awal menunjukkan bahawa mikroelektrod DEP dapat memanipulasi EV seperti yang dibuktikan oleh imej pendarfluor negatif dielektroforesis (NDEP). Kajian lebih lanjut mengenai analisis grid yang dijalankan menunjukkan ketekalan teori dan hasil yang dikemukakan. Nilai Jumlah Pendarflour Sel Betul (CTCF) daripada analisis grid menyimpulkan bahawa EV dimanipulasi ke pusat rantau tumpuan (ROI). Oleh itu, teknik DEP ini mencadangkan strategi cepat untuk pengasingan EV daripada mediumnya dalam jumlah yang sedikit di samping mengekalkan ketepatan dan keberkesanan kos.

Kata kunci: Dielektroforesis; manipulasi; pemisahan zarah; pengasingan; vesikel ekstrasel

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*Pengarang untuk surat-menyurat; email: muhdramdzan@ukm.edu.my