Intelligent optofluidic analysis for ultrafast single bacterium profiling of cellulose production and morphology

Yu, Jinqian; Sun, Guoyun; Lin, Weikang Nicholas; Vadanan, Sundaravadanam Vishnu; Lim, Sierin; Chen, Chia-Hung

doi:10.1039/c9lc01105f

Cited by 7 publications

(3 citation statements)

References 24 publications

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“…Quantum efficiency (QE) is an essential performance metric of optical sensors, as it expresses the ratio of incident photons to generated electrons [86,87]. CCDs are popular choices for droplet microfluidic devices due to their high light sensitivity, as indicated in [76,[88][89][90]. Although CCD sensors are widely used in DMFC, they are not ideal for high-throughput applications.…”

Section: Detection Setupsmentioning

confidence: 99%

Optical Detection Methods for High-Throughput Fluorescent Droplet Microflow Cytometry

et al. 2021

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High-throughput microflow cytometry has become a focal point of research in recent years. In particular, droplet microflow cytometry (DMFC) enables the analysis of cells reacting to different stimuli in chemical isolation due to each droplet acting as an isolated microreactor. Furthermore, at high flow rates, the droplets allow massive parallelization, further increasing the throughput of droplets. However, this novel methodology poses unique challenges related to commonly used fluorometry and fluorescent microscopy techniques. We review the optical sensor technology and light sources applicable to DMFC, as well as analyze the challenges and advantages of each option, primarily focusing on electronics. An analysis of low-cost and/or sufficiently compact systems that can be incorporated into portable devices is also presented.

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Section: Detection Setupsmentioning

confidence: 99%

Optical Detection Methods for High-Throughput Fluorescent Droplet Microflow Cytometry

et al. 2021

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“…12,13 Droplet-based hydrogel technology is designed to encapsulate the cells to determine their proliferation using flow cytometry. 14 The flow cytometry-based approach has been extended to integrate drop-screen technology for biomedical applications. 15−17 For the above advantages, most flow cytometry-based approaches are dependent on fluorescence readouts, and the fluorescence labeling process affects cell viability after screening.…”

mentioning

confidence: 99%

“…At present, two broad categories of methods are available for single stem cell analysis: flow cytometry and microwell-based platforms. − As a promising tool for single-cell screening with a throughput of ∼10 4 cells/min, flow cytometry is widely used to measure cell size, surface biomarkers and intracellular signal molecules. , Cellular antibody secretion is measured through the fluorescence labeling of cell-encapsulated hydrogel particles. , Droplet-based hydrogel technology is designed to encapsulate the cells to determine their proliferation using flow cytometry . The flow cytometry-based approach has been extended to integrate drop-screen technology for biomedical applications. − For the above advantages, most flow cytometry-based approaches are dependent on fluorescence readouts, and the fluorescence labeling process affects cell viability after screening. − The microwell-based platform has been developed to measure single-cell secretions and was designed specifically for function-driven single-cell screening. − However, the screening throughput of microwell-based approaches is limited by the number of microwells in one microtiter plate (∼100–1000 per experimental run) .…”

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confidence: 99%

Functional Stem Cell Sorting via Integrative Droplet Synchronization

et al. 2020

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Stem cell regenerative medicine strategy requires selecting functional cells to trigger repair processes. Stem cell secretion measurement is important to evaluate cellular activities for functional cell sorting. At present, to determine single cell secretions, mixing chemical sensors and cells together in a chamber is a standard procedure. However, toxic chemical sensors, such as albumin assay kits, are used during this process, causing low viability (64%) and low functionality (30%). It is especially important for stem cell profiling, as the toxicity of chemical sensors such as albumin permanently changes stem cell phenotypes, leading to unwanted analysis outcomes. Moreover, because of the sensor toxicity, the challenge of culturing sorted cells remain. In this study, an integrative synchronized droplet screen system was developed to separate a large droplet with cell encapsulation into two daughter droplets: one droplet containing cell secretions and the other droplet containing a single cell. These two daughter droplets moved along the channels at the same speed in synchronization. By injecting toxic chemical sensors into one daughter droplet, the single-cell secretions were determined without affecting the cells in the corresponding droplet. Based on the daughter droplet synchronization, the cells without mixing toxicity sensors were sorted for cell culturing. For example, to identify hepatocytes, the albumin secretion of undifferentiated HepaRG stem cells was measured in daughter droplets by injecting a toxic albumin assay kit for functional stem cell sorting. With synchronized sorting, functional hepatocytes were collected without exposure to toxic chemical sensors, showing high viability (78%) and active functionality (89%).

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Future foods: Design, fabrication and production through microfluidics

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et al. 2022

Biomaterials

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Intelligent optofluidic analysis for ultrafast single bacterium profiling of cellulose production and morphology

Abstract: A continuous-flow intelligent optofluidic device using a convolutional neural network (CNN) computational method was developed to enable high-throughput single-bacterium profiling of bacteria cellulose (BC) with a throughput of ∼35 bacteria per second.

Cited by 7 publications

References 24 publications

Optical Detection Methods for High-Throughput Fluorescent Droplet Microflow Cytometry

Optical Detection Methods for High-Throughput Fluorescent Droplet Microflow Cytometry

Functional Stem Cell Sorting via Integrative Droplet Synchronization

Future foods: Design, fabrication and production through microfluidics

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