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Artículo

Label-free counting of Escherichia coli cells in nanoliter droplets using 3D printed microfluidic devices with integrated contactless conductivity detection

Duarte, Lucas C.; Figueredo, FedericoIcon ; Vaz, Luiz Paulo Ribeiro; Corton, EduardoIcon ; Coltro, Wendell K.T.
Fecha de publicación: 04/2019
Editorial: Elsevier Science
Revista: Analytica Chimica Acta
ISSN: 0003-2670
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Química Analítica

Resumen

This study describes for the first time the development of 3D printed microfluidic devices with integrated electrodes for label-free counting of E. coli cells incorporated inside droplets based on capacitively coupled contactless conductivity detection (C4D). Microfluidic devices were fully fabricated by 3D printing in the T-junction shape containing two channels for disperse and continuous phases and two sensing electrodes for C4D measurements. The disperse phase containing E. coli K12 cells and the continuous phase containing oil and 1% Span® 80 were pumped through flow rates fixed at 5 and 60 μL min−1, respectively. The droplets with incorporated cells were monitored in the C4D system applying a 500-kHz sinusoidal wave with 1 Vpp amplitude. The generated droplets exhibited a spherical shape with average diameter of 321 ± 9 μm and presented volume of 17.3 ± 0.5 nL. The proposed approach demonstrated ability to detect E. coli cells in the concentration range between 86.5 and 8650 CFU droplet−1. The number of cells per droplet was quantified through the plate counting method and revealed a good agreement with the Poisson distribution. The limit of detection achieved for counting E. coli cells was 63.66 CFU droplet−1. The label-free counting method has offered instrumental simplicity, low cost, high sensitivity and compatibility to be integrated on single microfluidic platforms entirely fabricated by 3D printing, thus opening new possibilities of applications in microbiology.
Palabras clave: 3D PRINTING TECHNOLOGY , BIOANALYTICAL CHEMISTRY , CELLS SIZING , DROPLET-BASED MICROFLUIDICS , MICROBIOLOGY , SINGLE CELLS
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info:eu-repo/semantics/restrictedAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/123262
URL: https://linkinghub.elsevier.com/retrieve/pii/S0003267019304799
DOI: https://doi.org/10.1016/j.aca.2019.04.045
Colecciones
Articulos(ICT - MILSTEIN)
Articulos de INST.DE CS. Y TECNOLOGIA "DR. CESAR MILSTEIN"
Articulos(IQUIBICEN)
Articulos de INSTITUTO DE QUIMICA BIOLOGICA DE LA FACULTAD DE CS. EXACTAS Y NATURALES
Citación
Duarte, Lucas C.; Figueredo, Federico; Vaz, Luiz Paulo Ribeiro; Corton, Eduardo; Coltro, Wendell K.T.; Label-free counting of Escherichia coli cells in nanoliter droplets using 3D printed microfluidic devices with integrated contactless conductivity detection; Elsevier Science; Analytica Chimica Acta; 1071; 4-2019; 36-43
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