Abstract
Although fluorescence microscopy is the gold standard tool for biomedical research and clinical applications, their use beyond well-established laboratory infrastructures remains limited. The present study investigated a novel on-chip cell analysis platform based on contact fluorescence microscopy and microfluidics. Combined use of a contact fluorescence imager based on complementary metal-oxide semiconductor technology and an ultra-thin glass bottom microfluidic chip enabled both to observe living cells with minimal image distortion and to ease controlling and handling of biological samples (e.g. cells and biological molecules) in the imaged area. A proof-of-concept experiment of on-chip detection of cellular response to endothelial growth factor demonstrated promising use for the recently developed on-chip cell analysis platform. Contact fluorescence microscopy has numerous desirable features including compatibility with plastic microfluidic chips and compatibility with the electrical control system, and thus will fulfill the requirements of a fully automated cell analysis system.
Highlights
A wide variety of sophisticated fluorescent probes enables the high sensitivity and specificity of cellular activity detection
An integrated platform of microfluidics and on-chip microscopy has been intensively developed based on bright field imaging techniques.[15,16,17,18,19]
Implementation of contact fluorescence microscopy in a microfluidic chip platform is not simple, due to certain technical obstacles that arise from the characteristic features of fluorescence imaging
Summary
A wide variety of sophisticated fluorescent probes enables the high sensitivity and specificity of cellular activity detection. Conventional tabletop-type fluorescence microscopes are relatively bulky and expensive, due to the series of lenses and mechanical structures required for focusing lenses. Their use beyond well-established laboratory infrastructures with welltrained technicians remains limited.[1]. On-chip imaging techniques are highly attractive, in the application fields of automated systems, for high-throughput biological and medical screening due to their advantages compared with conventional tabletop-type microscopes, including (i) ease of scale-up of field of view with large numbers of pixels and smaller pixel sizes, (ii) compact hardware geometry, cost effectiveness and mass producibility, (iii) and simple usability without careful selection and focusing of the lenses.[2,3,4,5,6,7,8] Automated system using microfluidic technology has demonstrated great potential for biological and medical analysis,[9,10] drug screening,[11,12] and cell processing.[13,14] using on-chip fluorescence imaging techniques and a microfluidic chip platform provides a fully automated system from sample handling to detect of cellular activity
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