Abstract
SummaryIn a previous paper, we showed a new giant lens called the Mesolens and presented performance data and images from whole fixed and intact fluorescently‐stained 12.5‐day old mouse embryos. Here, we show that using the Mesolens we can image an entire Drosophila larva or adult fly in confocal epifluorescence and show subcellular detail in all tissues. By taking several hundreds of optical sections through the entire volume of the specimen, we show cells and nuclear details within the gut, brain, salivary glands and reproductive system that normally require dissection for study. Organs are imaged in situ in correct 3D arrangement. Imaginal discs are imaged in mature larvae and it proved possible to image pachytene chromosomes in cells within ovarian follicles in intact female flies. Methods for fixing, staining and clearing are given.
Highlights
Drosophila has been described as ‘too small for easy handling but too large for microscopy’ (Chyb & Gompel, 2013)
Wild-type Drosophila melanogaster were anaesthetized by placing in a plastic tube surrounded by dry ice for 2 min
In a typical imaging session, up to 220 confocal optical sections were obtained in the 850 μm thickness of the larva and adjacent sections showed quite different patterns of nuclei and even of cells, since the optical section thickness was less than 4 μm
Summary
Drosophila has been described as ‘too small for easy handling but too large for microscopy’ (Chyb & Gompel, 2013). The Mesolens, with its unusual combination of low magnification and high numerical aperture, solves the size problem without compromising image resolution (McConnell et al, 2016), but the impermeability of the cuticle prevents the use of many preparative methods. Light-sheet illumination has been successful in imaging specimens up to 400 μm long (Tomer et al, 2012). Other methods such as micro-CT (Matsuyama et al, 2015) and optical coherence tomography (McGurk et al, 2007) provide only low-resolution images
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