Spectral flow cytometry for crop cell autofluorescence characterization

Overview

This offering provides advanced cellular phenotyping services for crop plants using the latest generation of full-spectrum (spectral) flow cytometry. By characterizing the autofluorescence profiles of single cells (protoplasts) prepared from crop plant organs, the technology can identify phenotypic differences between individual cells of the same type and detect previously unknown cell types. These insights are directly relevant to engineering stress resilience and productivity in crops, supporting agricultural biotechnology innovation.

Technical specifications

Core technology:

• Full-spectrum/spectral flow cytometry using a Cytek Aurora instrument
• Detection of subtle changes in autofluorescence spectra from individual cells
• UMAP dimensionality-reduction analysis for clustering protoplasts based on autofluorescence similarity
• Spectral sorting capability enabling collection of specific cell populations for downstream molecular analysis

Key capabilities:

• Protoplast preparation from maize and other crop leaf peels and tissue types
• Identification of known leaf cell types including pavement, mesophyll, bundle sheath, guard, and subsidiary cells
• Detection of novel or unknown cell types based on autofluorescence signatures
• Analysis across different tissue types, developmental stages, and environmental conditions
• Compatibility with transgenic plants expressing markers in known cell types for validation

Research applications:

• Understanding cellular function, physiology, metabolism, and biochemistry
• Informing strategies for manipulating crop growth, development, and productivity
• Studying cellular responses to environmental stress conditions

Technology readiness level

The technology has been validated through initial experiments using maize (B73) leaf peels, where protoplast autofluorescence profiles were successfully characterized and clustered using UMAP analysis. Multiple distinct clusters were detected, hypothesized to represent known leaf cell types as well as potentially unknown cell types. Future validation plans include profiling protoplasts from diverse tissue types, developmental stages, and environmental conditions, as well as using transgenic marker lines to associate autofluorescence phenotypes with known cell types. The spectral sorting capability will enable detailed molecular characterization of both known and novel cell populations.