Research

I study the evolution of developmental mechanisms in insects. For my full publication list, please see my profile at Google Scholar or ResearchGate. These are projects I’ve worked on recently:

My collaborators include: Jordan Hoffmann and Chris Rycroft at the Harvard John A. Paulson School of Engineering and Applied Sciences, Steve DiNardo at the University of Pennsylvania School of Medicine, James Crall at the Harvard University Center for the Environment, and members of the Extavour Lab.


High-throughput live-imaging of embryogenesis

Recent advances in microscopy have made it possible to capture morphogenesis in 3D time-lapses with excellent temporal and spatial resolution. The datasets that result are quite useful, but they are time- and labor-intensive to generate. We show that for some experimental systems a complementary approach is also helpful, namely high-throughput epifluorescence. It is technically simple, and it allows the user to retain a comparable x-y and temporal resolution, while trading off excellent z-resolution for a 10- to 100-fold increase in sample size.

I was recently invited by Science/AAAS to present on this work for an online microscopy webinar. My co-presenters were Dr. Ed Boyden from MIT and Dr. Meltsje de Hoop from Sanofi S.A.

The hour-long webinar covers several ways that biologists can add automation to their microscopy-based research. The video recording of the webinar is available for free, but viewers need to register to get access:

de Hoop, M., Donoughe, S., and Boyden, E. (2017). What automation can do for you: The benefits and pitfalls of automating your microscopy research. Science, 355.6323, 421-421. (link to free webinar; slides also available for download)

While developing techniques for time-lapse imaging, I also had the opportunity to beta-test the Zeiss Celldiscoverer 7, a microscope that was designed specifically for high-throughput live-imaging. I worked with Sebastian Gliem at Zeiss, and we co-wrote an article for the industry magazine Imaging and Microscopy:

Donoughe, S. and Gliem, S. (2016). High-Throughput, Long-Term Live Imaging: Automated Microscopy of Insect Development. Imaging and Microscopy. (PDF)


Modular, customizable molds for mounting samples

Live-imaging embryos in a high-throughput manner is essential for shedding light on a wide range of questions in developmental biology, but it is difficult and costly to mount and image embryos in consistent conditions. We are developing tools to make it easier for biologists to design and build their own custom mounting devices.

A forthcoming journal article will describe these devices in detail, but in the mean time please don’t hesitate to contact me for more information about the designs or protocols (donoughe [at] harvard.edu). A pre-print is available on bioRxiv:

Donoughe, S., Kim, C., & Extavour, C. G. (2017). High-throughput live-imaging of embryos in microwell arrays using a modular, inexpensive specimen mounting system. bioRxiv. (PDF) (link to bioRxiv)

Photo credits: beta-testers Andrew Gehrke and Mara Laslo


Embryonic development of the cricket Gryllus bimaculatus

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SUMMARY: The two-spotted field cricket, Gryllus bimaculatus is an increasingly popular model species. It is a member of the order Orthoptera, a group that branches basally with respect to the clade that includes flies, beetles, bees, and butterflies. We used a combination of brightfield timelapse microscopy, confocal microscopy, 3D reconstructions, and measurements of egg dimensions to comprehensively describe the morphogenesis and anatomy of the cricket embryo. This resource will facilitate further studies on G. bimaculatus development and serve as a useful point of reference for other studies of wild-type and experimentally manipulated insect development.

Donoughe, S., & Extavour, C. G. (2016). Embryonic development of the cricket Gryllus bimaculatus. Developmental Biology, 411(1), 140-156. (PDF) (link to journal)

NOTE: On the Resources page of this website, I have included additional materials for anyone interested in cricket embryonic development.

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The role of BMP signaling in cricket germ cell specification

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SUMMARY: Primordial germ cells are the cells that turn into sperm or egg. Thus, they are the only lineage of cells in a body that give rise to the next generation. For the first time, we uncovered the molecular basis of germ cell formation in an insect other than fruit fly, discovering that the process is strikingly similar to that of mice.

Donoughe, S., Nakamura, T., Ewen-Campen, B., Green, D. A., Henderson, L., & Extavour, C. G. (2014). BMP signaling is required for the generation of primordial germ cells in an insect. Proceedings of the National Academy of Sciences, 111(11), 4133-4138. (PDF) (link to journal)
Ewen-Campen, B., Donoughe, S., Clarke, D. N., & Extavour, C. G. (2013). Germ cell specification requires zygotic mechanisms rather than germ plasm in a basally branching insect. Current Biology, 23(10), 835-842. Recommended by the Faculty of 1000 (PDF) (link to journal)
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Planar polarity in the epidermis of Drosophila melanogaster

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SUMMARY: In order to function properly, cells often need to “know” things beyond the information encoded in their genome. One thing a cell needs to know is how it is oriented with respect to the arrangement of the other cells in its tissue; this is called cell polarity. In this project we used the epidermis of the fruit fly to ask how different genes contribute to cell polarity. To our surprise, we discovered that two groups of genes separately imbue cells with the same sort of cell polarity, but each group has a particular subset of cells where its effect is strongest.

Donoughe, S., & DiNardo, S. (2011). dachsous and frizzled contribute separately to planar polarity in the Drosophila ventral epidermis. Development, 138(13), 2751-2759. Recommended by the Faculty of 1000 (PDF) (link to journal)
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The biomechanics of dragonfly and damselfly wing flexibility

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SUMMARY: How could a dragonfly change the shape of its wings during flight even though it has no muscles within the wings? We used mechanical tests and detailed micrographs of 12 species to show that tiny flexible protein pads and mechanical linkages are built into the surface of wings. These microscopic structures affect how a wing changes shape in response to aerodynamic forces.

Donoughe, S.*, Crall, J. D.*, Merz, R. A., & Combes, S. A. (2011). Resilin in dragonfly and damselfly wings and its implications for wing flexibility. Journal of morphology, 272(12), 1409-1421. (PDF) (link to journal)
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