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Barcode specific immobilization interface for microfluidics-assisted spatial analysis microarray uniform barcoding
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DOI：10.1039/D5AN00534E.png)
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Abstract
En 中文
Microfluidics-assisted spatially barcoded microarray technology offers a high-throughput; low-cost approach to spatial transcriptomic profiling. A uniform barcoded microarray is crucial for spatially unbiased mRNA analysis. However; non-specific adsorption of barcoding reagents in microchannels occurs during liquid transport; causing nonuniform barcoding in chip functional regions. The uneven microarray further leads to biased transcriptome capture. Herein; we develop a barcode specific immobilization (BarSI) interface with both anti-adsorption property and biological activity for the development of uniform spatially barcoded microarray chips. We immobilize DNA probes in straight and serpentine microchannels with coefficient of variation (CV) of 2.3% and 3.2%. Based on the orthogonal barcoding system; we develop spatially barcoded microarray chips with an overall fluorescence intensity CV of 8.47 ± 1.26%; compared with the CV of 20.91 ± 2.84% of microarrays developed on conventional amino glass slides. Using the uniform spatially barcoded microarray chip; we achieve spatially unbiased detection of mouse liver mRNA with the absolute value of Moran’s I below 0.05. We present an economical and accessible method for manufacturing uniform spatially barcoded microarray chips; introducing a novel strategy for unbiased transcriptome analysis.
Keywords:
microfluidics
spatial transcriptomics
barcoded microarray
uniform immobilization
transcriptome analysis