Development of simple and efficient Lab-on-a-Disc platforms for automated chemical cell lysis

Arash Khorrami Jahromi,Maryam Saadatmand,Laleh Parsa Yeganeh,Manouchehr Eghbal

doi:10.1038/s41598-020-67995-3

Abstract

Cell lysis is the most important first step for molecular biology and diagnostic testing. Recently, microfluidic systems have attracted considerable attention due to advantages associated with automation, integration and miniaturization, especially in resource-limited settings. In this work, novel centrifugal microfluidic platforms with new configurations for chemical cell lysis are presented. The developed systems employ passive form of pneumatic and inertial forces for effective mixing of lysis reagents and cell samples as well as precise fluidic control. Characterizations of the developed Lab-on-a-Discs (LoaDs) have been conducted with dyed deionized (DI) waters and white blood cells (WBCs) to demonstrate the suitability of the proposed systems in terms of mixing, fluidic control and chemical cell lysis. By making comparison between the results of a well-established manual protocol for chemical cell lysis and the proposed chemical cell lysis discs, it has been proved that the developed systems are capable of realizing automated cell lysis with high throughput in terms of proper values of average DNA yield (ranging from 20.6 to 29.8 ng/µl) and purity (ranging from 1.873 to 1.907) as well as suitability of the released DNA for polymerase chain reaction (PCR). By considering the manual chemical lysis protocol as a reference, the efficiency of the LoaDs has been determined 95.5% and 91% for 10 min and 5 min lysis time, respectively. The developed LoaDs provide simple, efficient, and fully automated chemical cell lysis units, which can be easily integrated into operational on-disc elements to obtain sample-to answer settings systems.

Highlights

Cell lysis is the most important first step for molecular biology and diagnostic testing
Heating up to high temperature can cause damage to facilities or exposed biological elements[11]. Another practical method is chemical cell lysis, in which cell membranes are disrupted by adding genuine reagents to the samples
Buoyancy driven bubble was employed for mixing a cell sample and lysis reagents to perform chemical lysis on a LoaD platform

Summary

Introduction

Cell lysis is the most important first step for molecular biology and diagnostic testing. For implementing the chemical lysis method on centrifugal microfluidic platforms, design and fabrication of units for efficient mixing as well as precise fluidic control of samples are required. All these requirements can be readily achieved by directly designing the units on a LoaD, independent of complicated and external accessories. High cell lysis efficiency has been reported for the system, and the it has a novel mechanism and configuration, complexity of design and fabrication of the system may hamper further developments and integration of the proposed lysis unit into other on-disc units for achieving a sample-to-answer s ystem[13]. The systems rely heavily upon external accessories, which may imposes an additional cost/complexity on performing cell lysis protocol 11,15

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