Deep-tissue Imaging using Selective Plane Illumination Microscopy in Drosophila

Abstract

The imaging speed and penetration depth limit the study of dynamics of the lateral mesodermal cells during gastrulation in Drosophila. Selective Plane Illumination Microscope(SPIM) is an important tool to image the rapid cell shape changes on the whole-organism level. However, both illumination light and fluorescence signal are strongly scattered in the fly embryos which reduces the contrast in images. To improve the deep-tissue imaging using SPIM, we developed new membrane markers with fluorophores that emit in the far-red and near-infrared region of electromagnetic spectrum. Also, we used deconvolution to improve the contrast in the acquired images by computationally reducing the noise and blur. Use of fluorophores emitting at longer wavelengths and deconvolution helps in producing more interpretable images with high signal-to-noise ratio at greater depths. Such images will facilitate the image analysis and thus generate reliable 3D reconstructions of cells which will be essential in quantitative analyses of shape changes in the lateral mesodermal cells.