PySeq2500

PySeq2500 repurposes Illumina HiSeq 2500 systems to perform automated fluidics control and four‑channel fluorescence imaging for iterative indirect immunofluorescence (4i) and related multiplexed fluorescence microscopy workflows.

Key Features:

• Open Platform: An open-source Python codebase provides programmable control of HiSeq 2500 hardware.
• Flow Cell Design: A custom flow cell design integrates with HiSeq 2500 hardware to enable fluidics and imaging functions.
• Temperature Control and Reagent Exchange: Precise temperature regulation and automated reagent exchange (fluidics) for multi-step protocols.
• 4-Channel Image Acquisition: Simultaneous acquisition across four imaging channels for multiplexed fluorescence data.
• Stable Positioning: Maintains consistent sample positioning throughout extended experiments.

Scientific Applications:

• Iterative indirect immunofluorescence imaging (4i): Unattended execution of staining, imaging, and antibody elution cycles using commercially available antibodies.
• Multiplexed fluorescence microscopy over multi-day experiments: Generation of robust, reproducible multiplexed datasets across extended experimental timelines.
• Spatial profiling of tissue: Mapping cell types and pathological features in samples such as mouse and postmortem human spinal cord sections.

Methodology:

An open-source Python codebase automates HiSeq 2500 fluidics, temperature regulation, stage positioning, and four-channel image acquisition.