# Protocol for 3D surface texture modeling and quantitative spectral decomposition analysis in Drosophila border cell clusters

## TL;DR

This paper presents a protocol to model 3D surface textures of Drosophila border cell clusters and analyze how genes affect their geometry.

## Contribution

A new method for 3D surface modeling and spectral decomposition analysis of cell clusters using Airyscan microscopy.

## Key findings

- Airyscan images can be converted into 3D surface models showing convex and concave curvature.
- Spectral decomposition analysis reveals how genes influence surface texture differences across genotypes.
- The protocol is applicable to Drosophila border cells and potentially other cell types.

## Abstract

Drosophila border cell clusters model collective cell migration. Airyscan super-resolution microscopy enables fine-scale description of cluster shape and texture. Here we describe how to convert Airyscan images of border cell clusters into 3D models of the surface and detect regions of convex and concave curvature. We use spectral decomposition analysis to compare surface textures across genotypes to determine how genes of interest impact cluster surface geometry. This protocol applies to border cells and could generalize to additional cell types.

For complete details on the use and execution of this protocol, please refer to Gabbert et al.1

•Drosophila cell-specific transgene expression and ovary dissection and fixation•Airyscan image acquisition of border cell clusters with high-resolution z stacks•Extraction of cluster surface to generate 3D models of the surface shape and texture•Spectral decomposition analysis to quantify differences in surface geometry

Drosophila cell-specific transgene expression and ovary dissection and fixation

Airyscan image acquisition of border cell clusters with high-resolution z stacks

Extraction of cluster surface to generate 3D models of the surface shape and texture

Spectral decomposition analysis to quantify differences in surface geometry

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Drosophila border cell clusters model collective cell migration. Airyscan super-resolution microscopy enables fine-scale description of cluster shape and texture. Here we describe how to convert Airyscan images of border cell clusters into 3D models of the surface and detect regions of convex and concave curvature. We use spectral decomposition analysis to compare surface textures across genotypes to determine how genes of interest impact cluster surface geometry. This protocol applies to border cells and could generalize to additional cell types.