Optimization of fluorescent cell-based assays for high-throughput analysis using microchamber array chip formats

Abstract

This study describes the optimization of fluorescent cell-based assays using microchamber array chip formats as well as using automatic nanoliter volumes of sample dispensing system for high-throughput screening analysis of anticancer drugs. Cell-based assays can be employed efficiently in the screening of potential anticancer drug candidates and bioactive compounds with distinct biological function. Identification and development of cell-based assays adapted to high-throughput screening requirements is important when screening chemicals for their potential anticancer properties. Cell-based screening assays using microchamber array chip formats and automatic nanoliter volumes of sample dispensing system requires an optimization as a prerequisite for parameters including assay liquid volume and number of cells per each chamber, and the total cell-based assay itself. Further, the anticancer effect of mitomycin C was studied as an example against human cervical carcinoma cell line-HeLa 229 using cell-based assay that was optimized on chamber array chip formats and determined the cytotoxicity of mitomycin C by measuring the cell proliferation of HeLa with Calcein-AM fluorescent dye. The cell-based screening assay that was performed using chamber array chip formats was compared with the conventional 96-well plate formats was discussed. The assay described in this study is rapid, simple and inexpensive that is desirable in selecting anticancer candidates. q

Cellular heterogeneity and live cell arrays

“…Microwell chips have been fabricated using various lithography techniques, including X-ray lithography, scanning probe lithography, and photolithography. 178,190,191 These techniques can be coupled with additional etching and printing procedures for further control of array parameters (Fig. 13), such as well dimensions and overall array size.…”

dPCR: A Technology Review

“…This in turn necessitates a method that isolates microwells from one another and avoids rapid evaporation of minuscule volumes. To support efficient microwell filling and partitioning, it is necessary to have differential surface properties between the interior of the microwell that needs to be hydrophilic and the top face of the array (in between the microwells) that needs to be hydrophobic [ 58 , 59 , 60 ].…”

Cell chips as new tools for cell biology – results, perspectives and opportunities

“…A sensitive fluorescent microarray scanner was employed to measure cell proliferation exploiting calcein-AM dyes to study the anticancer effect of mitomycin C against a human cervical carcinoma cell line (HeLa). 31 A simple and smart microfluidic PDMS device was instead realized by Cira et al to identify the minimum inhibitory concentration of antibiotics on bacterial population. 61 In this case, a bacterial suspension was introduced in a set of interconnected chambers preloaded with different antibiotics or antibiotic concentrations; growth and viability were then detected by a colorimetric change thanks to a pH indicator (phenol red).…”