UMGC white paper: Automated nuclei isolation from diverse plant tissues using the Singulator 200
The application of single-cell sequencing technologies has revolutionized developmental biology; however, plant genomics has lagged behind due to the structural barrier of the cell wall. Traditional protoplasting methods are often labor-intensive, species-specific, and prone to introducing transcriptional stress artifacts. Single-nucleus RNA sequencing (snRNA-seq) offers a robust alternative but requires efficient, high-quality nuclei isolation, which remains technically challenging for fibrous or recalcitrant plant tissues.
This whitepaper evaluates the feasibility of the Singulator 200 to automate and standardize nuclei isolation across diverse plant species, including pennycress, soybean, corn, and tobacco. The UMGC compared yield, viability, and morphological integrity of nuclei isolated from fresh and frozen leaf, root, and flower tissues.
Results indicate successful nuclei recovery from multiple plant tissue types. Representative results from pennycress leaf tissue are shown below, demonstrating consistent yields across input masses and comparable performance between fresh and frozen samples.
| Condition | Plant Age (days) | Input Mass (mg) | Total Yield (nuclei) |
|---|---|---|---|
| Fresh | ~28 days | 50 | 6.57e5 |
| Fresh | ~28 days | 150 | 9.67e5 |
| Frozen | ~28 days | 50 | 9.67e5 |
| Frozen | ~28 days | 150 | 1.07e6 |
Table 1: Nuclei yield from pennycress leaf tissue
Overall, the Singulator 200 represents a viable solution for researchers seeking to perform scalable plant single-cell nuclei isolations. This study outlines a validated workflow for generating high-quality plant nuclei suitable for downstream applications such as 10x Genomics snRNA-seq.
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