A novel two-layer-integrated microfluidic device for high-throughput yeast proteomic dynamics analysis at the single-cell level

Abstract

Current microfluidic methods for studying multicell strains (e.g., m-types) with multienvironments (e.g., n-types) require large numbers of inlets/outlets (m*n), a complicated procedure or expensive machinery. Here, we developed a novel two-layer-integrated method to combine different PDMS microchannel layers with different functions into one chip by a PDMS through-hole array, which improved the design of a PDMS-based microfluidic system. Using this method, we succeeded in converting 2 × m × n inlets/outlets into m + n inlets/outlets and reduced the time cost of loading processing (from m × n to m) of the device for studying multicell strains (e.g., m-types) in varied multitemporal environments (i.e., n-types). Using this device, the dynamic behavior of the cell-stress-response proteins was studied when the glucose concentration decreased from 2% to a series of lower concentrations. Our device could also be widely used in high-throughput studies of various stress responses, and the new concept of a multilayer-integrated fabrication method could greatly improve the design of PDMS-based microfluidic systems.

Quantitative proteomics characterization of cancer biomarkers and treatment

“… 111 As a topic of frequent discussion in the past decade, single-cell proteomics evaluates the heterogeneity and rare types of cells based on cell types and the state of a single cell. 112 , 113 Multi-omics approaches have become promising in the study of human diseases. 60 , 61 , 62 HSP90β was identified as a potential prognostic biomarker for lung cancer through integrative analysis of proteome, phosphoproteome, transcriptome, and whole-exome sequencing data.…”

Cultivating Multidisciplinarity: Manufacturing and Sensing Challenges in Cultured Meat Production

“…Nowadays, advanced microfluidics integrates a number of operations into a single chip, such as sample pre-treatment and preparation, DNA extraction, amplification, separation and mixing of the samples, a micromechanical system for fluid manipulation, together with optical and electronic components for signal sensing. The microfluidic technology has been widely used to study cell biology for biomedical applications [88,89], protein studies [90], pathogen detection [91,92], cell culture [93,94] or tissue engineering [95]. The dimensions of microfluidic channels and the physical scale of cells correspond to each other.…”

Two-Layered Microfluidic Devices for High-Throughput Dynamic Analysis of Synthetic Gene Circuits in E. coli

“…Here, we designed a two-layered microfluidic device, which was combined with two PDMS layers with different functions. One of the layers was designed for the bacterial culture, and the other was designed as the sample inlet.…”

“…Compared with larger cells, the prevention of cross contamination in E. coli can be more difficult. Small fence structures 21,30 that are routinely used to prevent cross contamination can no longer be applicable. For the characterization of synthetic gene elements such as activators or repressors, multiple concentrations of inducers are needed to derive accurate activation or repression functions.…”