Automatic Microfluidic Cell Wash Platform for Purifying Cells in Suspension: Puriogen.

Cell preparation is a critical step in many biological assays. Traditional centrifugation-based washing methods are widely used but come with limitations. These include high labor demands, time consumption, and significant cell loss. Prior research has shown that manual pipetting leads to inconsistent results and poor sample purity. This gap motivated the development of automated alternatives. No prior work had resolved the issue of high-throughput, centrifuge-free cell washing. Existing methods fail to address the need for rapid and efficient cell purification. The lack of a reliable microfluidic solution has hindered progress in single-cell sequencing. This paper introduces a novel platform to overcome these limitations.

The goal of this work is to develop a centrifuge-free cell washing platform. The aim is to improve cell purity without manual intervention. The study seeks to address the inefficiencies of traditional centrifugation methods. A key objective is to reduce labor and time while maintaining high cell viability. The platform is intended to support downstream analyses like single-cell sequencing. The researchers propose an automated solution using microfluidics. The study aims to demonstrate high-throughput processing capabilities. The purpose is to provide a reliable and efficient cell preparation alternative.

The Puriogen platform uses an inertial microfluidic device for cell washing. The design is based on inertial focusing principles to separate cells from contaminants. The system is fully automated and does not require centrifugation. The device is optimized for high-throughput processing of cell suspensions. The platform handles sample volumes at a rate exceeding 500 μL/min. It integrates fluidic control and sample processing in a single unit. The method is validated using fluorescent dye removal and cell viability tests. The system is tested for its ability to concentrate and purify cell suspensions.

Puriogen removes over 90% of ambient proteins and nucleic acids from cell samples. It reduces residual fluorescent dye by more than 90%, lowering background signals. The platform increases the live cell percentage in the sample by two times. It also enhances the single-cell population by dissociating small-cell aggregates. The device achieves a 20% increase in singlet cell concentration. Puriogen can concentrate cells fivefold in a single flow-through process. The system processes samples at a rate above 500 μL/min. These results suggest improved efficiency and purity compared to traditional methods.

The Puriogen platform offers a centrifuge-free cell washing solution. It achieves high-throughput processing with excellent wash efficiency. The system reduces manual labor and time while maintaining cell viability. The platform effectively removes contaminants and fluorescent dye. It increases the proportion of live and single cells in the sample. The device is suitable for single-cell sequencing and other downstream applications. The results suggest that Puriogen is a reliable alternative to traditional methods. The authors propose that this platform supports broader cell preparation needs.