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Related Experiment Video

Updated: Jan 18, 2026

Author Spotlight: Enhancing Drug Discovery - Development of Automated, Standardized Protocols for Nuclei Extraction from Frozen Tissues

Author Spotlight: Enhancing Drug Discovery - Development of Automated, Standardized Protocols for Nuclei Extraction from Frozen Tissues

Published on: July 28, 2023

An improved nuclei isolation protocol from leaf tissue for single-cell transcriptomics.

Gabriela Madrid^1,2, Gabriel Angelo Saraiva Raimundo², Fabian Andres Reyes²

¹Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida, United States of America.

|September 10, 2025

Summary

This summary is machine-generated.

Researchers developed a new method to isolate nuclei from maize leaves for single-nuclei RNA sequencing (snRNA-seq). This technique effectively removes chloroplast contamination, improving data quality for plant biology studies.

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Area of Science:

Plant Biology
Molecular Biology
Genomics

Background:

Traditional plant biology research focused on tissue, organ, or whole plant levels.
Single-cell transcriptomics, including single-nuclei RNA sequencing (snRNA-seq), offers insights into individual cell expression profiles.
Isolating intact nuclei is crucial for successful snRNA-seq, but challenging in chloroplast-rich tissues like maize leaves.

Purpose of the Study:

To develop an improved method for isolating nuclei from Zea mays (maize) leaves for snRNA-seq.
To reduce chloroplast contamination during nuclei isolation, which interferes with DAPI staining and sorting.
To enhance the quality of transcriptional profiling from maize leaf tissues.

Main Methods:

Developed an alternative nuclei isolation protocol for Zea mays leaves.
Utilized Fluorescent-Activated Cell Sorting (FACS) with modifications to remove chloroplasts.
Leveraged chloroplast autofluorescence for selective removal during the sorting process.

Main Results:

Achieved significantly reduced chloroplast contamination in isolated nuclei.
Demonstrated improved read alignment to the genome and transcriptome.
Obtained higher quality data for downstream snRNA-seq analysis.

Conclusions:

The enhanced protocol provides a straightforward solution for snRNA-seq in chloroplast-rich plant tissues.
This method facilitates deeper understanding of cell-to-cell gene expression heterogeneity in maize.
Enables new research avenues in plant molecular biology and developmental studies.