Tool for physically constricting tissues
In previous research, I experimentally tested some theory-based hypotheses by physically manipulating the shape and size of tissues during development. In order to do so in a more precise and repeatable manner, I designed and built a custom device. The design files and usage instructions are available for others to use. For detailed step-by-step instructions, see the supplemental materials of this paper.
Most of the components were laser-cut from acrylic. The design file for those pieces is available to download here.
Customizable molds for live-imaging tissues
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. I developed some simple tools to make it easier for biologists to design and build their own custom mounting devices.
This paper describes a simple, reusable device to easily mount dozens of embryos in arrays of agarose microwells with customizable dimensions and spacing.
It can be configured to mount specimens for upright or inverted microscopes, and includes a reservoir to hold live-imaging medium to maintain constant moisture and osmolarity of specimens during time-lapse imaging.
All of the device components can be fabricated by cutting pieces from a sheet of acrylic using a laser cutter or by making them with a 3D printer. We demonstrate how to design a custom mold and use it to live-image dozens of embryos at a time. We include descriptions, schematics, and design files for 13 additional molds for 9 animal species, including most major traditional laboratory models and a number of emerging model systems:
- annelid worm Capitella teleta
- zebrafish Danio rerio
- fruit fly Drosophila melanogaster
- frog Eleutherodactylus coqui
- cricket Gryllus bimaculatus
- panther worm Hofstenia miamia
- amphipod Parhyale hawaiensis
- red flour beetle Tribolium castaneum
If you’d like to make your own, here are some things to get you started:
- Descriptions of molds for each species (PDF)
- Design files for all components for laser cutting & etching (zipped file of PDFs and DXFs)
- Assembly instructions for the OMMAwell device (PDF)
- Additional mold insert files—one per species listed above—suitable for 3D printing instead of laser etching (zipped file of SLTs)
And here is an open-access paper for the project:
Using the molds for high-throughput, long-term, fluorescence microscopy
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 showed 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 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:
While developing techniques for time-lapse imaging, I also had the opportunity to beta-test the Zeiss Cell Discoverer 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: