Lossless Altered Histone Modification Analysis System (LAHMAS)

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Biology

Background:

• Miniaturizing biological assays with microfluidics offers enhanced sensitivity and throughput but faces challenges like evaporation and analyte loss.
• Exclusive Liquid Repellency (ELR) addresses these issues by using an oil phase to protect aqueous volumes and prevent surface binding.
• CUT&Tag is a complex multistep assay that can benefit from miniaturization for improved efficiency and reduced sample requirements.

Purpose of the Study:

• To develop a novel microfluidic platform for miniaturizing the CUT&Tag assay.
• To leverage Exclusive Liquid Repellency (ELR) and Exclusion-based Sample Preparation (ESP) for lossless liquid handling in microfluidic systems.
• To create a system capable of accurate chromatin profiling from low input and rare cell samples.

Main Methods:

• Developed a microfluidic platform using PDMS-silane treated glass surfaces immersed in silicone oil.
• Integrated Exclusive Liquid Repellency (ELR) and Exclusion-based Sample Preparation (ESP) principles.
• Utilized the platform for miniaturized CUT&Tag assays, named the Lossless Altered Histone Modification Analysis System (LAHMAS).

Main Results:

• The LAHMAS platform demonstrated lossless liquid handling and prevented sample evaporation.
• Effective processing of cell inputs as low as 100 cells was achieved.
• Higher specificity compared to macroscale CUT&Tag assays was observed, enabling accurate chromatin profiling.

Conclusions:

• The novel microfluidic platform effectively miniaturizes the CUT&Tag assay, overcoming common challenges.
• LAHMAS facilitates accurate chromatin profiling of low input and rare cell samples with enhanced specificity.
• This technology holds potential for advancing biological assays in various research applications.