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Publication Number
US-12553805-B2
Publication Date
2026-02-17
Expiration Date
Abstract
Provided herein are methods of preserving a biological sample on a first substrate, the methods including: (a) disposing a biological sample on a first substrate; (b) delivering a spacer agent to the first substrate, wherein the spacer agent is applied around the biological sample on the first substrate, thereby generating a chamber around the biological sample; (c) delivering a mounting agent to the chamber; and (d) assembling the first substrate with a second substrate, thereby preserving the biological sample on the first substrate.
Core Innovation
The invention provides a method of preserving a biological sample on a first substrate by disposing the biological sample on the first substrate and generating a chamber around the biological sample. A mixture comprising a blocking paste and a plurality of beads is delivered around the biological sample on the first substrate, where the plurality of beads comprises beads approximately uniform in volume and/or shape. The bead-and-blocking-paste mixture is applied to form the chamber around the biological sample on the first substrate.
A mounting agent is delivered to the chamber, and the first substrate is assembled with a second substrate to preserve the biological sample on the first substrate. The chamber is sealed by assembling the first substrate with a second substrate, thereby maintaining the biological sample in the sealed chamber. The preservation approach is further connected to downstream analysis in which the chamber-forming mixture can be removed by applying heat.
In a preserving and analyzing workflow, the method applies heat to the first substrate to partially or completely melt the mixture comprising the blocking paste and the plurality of beads and to remove the second substrate and the mixture from the first substrate. After removing the chamber-forming materials, the method contacts the first substrate with an array comprising a plurality of capture probes. The capture probes include a spatial barcode and a capture domain, and the method performs spatial transcriptomic analysis.
Claims Coverage
The partial set contains two independent claims. Across these claims, the inventive coverage is centered on forming a sealed chamber from a blocking paste and approximately uniform beads on a first substrate, preserving the biological sample via assembly with a second substrate, and enabling spatial transcriptomic analysis using capture probes with spatial barcodes and capture domains after heat removal of the chamber-forming mixture.
Bead and blocking paste chamber formation for sample preservation
disposing a biological sample on a first substrate; delivering a mixture comprising a blocking paste and a plurality of beads to the first substrate around the biological sample, thereby generating a chamber around the biological sample, wherein the plurality of beads comprises beads approximately uniform in volume and/or shape
Mounting agent and sealing by first and second substrate assembly
delivering a mounting agent to the chamber; assembling the first substrate with a second substrate, thereby preserving the biological sample on the first substrate
Heat melting and removal of chamber materials
applying heat to the first substrate, thereby partially or completely melting the mixture comprising the blocking paste and the plurality of beads and removing the second substrate and the mixture comprising the blocking paste and the plurality of beads from the first substrate
Spatial transcriptomic capture probe array with spatial barcode and capture domain
contacting the first substrate with an array comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises (i) a spatial barcode and (ii) a capture domain; performing spatial transcriptomic analysis
The independent claims collectively cover using a blocking paste mixed with beads having approximately uniform volume and/or shape to generate a chamber around a biological sample on a first substrate, delivering a mounting agent and sealing with a second substrate to preserve the sample, and applying heat to melt and remove the chamber materials followed by contacting an array of capture probes comprising spatial barcodes and capture domains to perform spatial transcriptomic analysis.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Spatial transcriptomic analysis using capture probes containing spatial barcodes and capture domains.
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