Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Staining and mounting of the Hibiscus trionum petals at early stages of development (A) Early-stage floral bud at the final dissection step, after propidium iodide staining (red color). The double arrow indicates the dissection site. Scale bar, 100 μm. (B) Petals being mounted in a drop of Hoyer’s medium. (C) Coverslip placed over the petals and gently to spread the medium evenly. (D) Example slide showing EdU-labeled petals. (E) Zoom view of the coverslip region of panel D. (F) Higher magnification of selected petals prepared for imaging. Scale bar, 1 mm.

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Staining, Dissection, Labeling, Imaging

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Imaging EdU-labeled Hibiscus trionum petals at early stages of development to visualize the repartition of cell division events across the adaxial epidermis (A) Slide positioned under an upright confocal microscope using a Plan-Apochromat 10×/0.45 objective lens. (B) Example confocal image showing EdU-labeled nuclei (green) and propidium iodide-stained membranes (red) in the adaxial epidermis of early-stage petals. Newly synthesized DNA is visualized via fluorescent nucleotide analog 5-ethynyl-2′-deoxyuridine (EdU). Scale bar: 100 μm.

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Imaging, Labeling, Microscopy, Staining, Synthesized

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Identifying EdU-labeled nuclei using spot detection (A) Step 1/3 -Initial parameter setup for spot detection. (B) Step 2/3. Channel selection and parameter adjustment for EdU-labeled nuclei. For Step 3/3, see .

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Labeling, Selection

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Select EdU-labeled nuclei of the central proximo-distal stripe for filtered spot analysis (Step 3/3) (A) Application of a quality filter (“Quality above automatic threshold”) to detect EdU-labeled nuclei. (B) Stripe selection by X-position. The detected nuclei are further filtered using the histogram slider to isolate a central stripe. This step enables quantitative analysis of nuclei relative position along the petal proximo–distal axis.

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Labeling, Selection

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Create your Scatter plot of the Edu-labeled nuclei position across the proximo-distal axis of the petal (A) Select region of interest. (B) Plot settings. (C) Exported final scatter plot.

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Labeling

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Labeling

Journal: STAR Protocols

Article Title: Protocol for spatial quantitative analysis of cell division events across the epidermis of developing Hibiscus trionum petals

doi: 10.1016/j.xpro.2026.104354

Figure Lengend Snippet: Analysis of the spatial distribution of EdU-labeled nuclei along the proximodistal axis of the petal (A) Example of an Excel file exported from Imaris, containing XYZ positions of EdU-labeled nuclei. (B) Structure of the formatted R input file (‘R_Results’ sheet), including six key columns required for analysis: Genotype, Stage, Petal, Y (nuclei position), L (petal length), and YNorm (normalized position). (C) Calculation of the relative position of each EdU-labeled nucleus by normalizing its Y coordinate to the total petal length (YNorm = Y/L). (D) Output from script section #1: Density plots of EdU-labeled nuclei per individual petals. (E) Output from script section #2: Pooled density plots of EdU-labeled nuclei per genotype.

Article Snippet: Example of output table showing EdU-labeled nuclei coordinates extracted from Imaris

Techniques: Labeling