A pragmatic approach succeeds at going green
It’s important to remember that lab sustainability is both good for the environment and for scientific productivity. Dr. Prober is characterized by his team as a highly pragmatic principal investigator - and that trait pervaded his group’s efforts to be certified through the Caltech Green Labs Network. A pragmatic lab group is driven to discover achievable improvements, rather than be distracted by wishful thinking. The focus on actionable ideas propels successful environmental stewardship.
The Prober lab investigates genetic and neuronal mechanisms which underlie sleep, with an emphasis on disordered sleep, a behavior with long-term consequences to human health, including chronic disease. By going green as a team, this neuroscience lab examined how their research consumed materials and energy. Their eyes were open to an opportunity for greener biochemistry.
Our thanks to research technician Tasha Cammidge for the following interview on targeting greener chemistry while pursuing the elusive nature of sleep-wake states with genetic, pharmacological, and neuronal approaches.
a sustainability mindset leads to a solvent switch
Courtesy of the Prober Lab (Lee et al., 2019). A) Schematic of 5-dpf zebrafish (Danio rerio) brain. A, anterior; L, lateral; V, ventral; Ce, cerebellum; Hy, hypothalamus; TeO, tectum. D,D’) in situ hybridization (ISH) of transforming growth factor, alpha (tgfa) coexpression with transcription factor sox2 in 5-dpf zebrafish brains. Coexpression indicated by white arrowheads. Dashed line in (D) shows outline of brain. Scale bar 30 mm.
What is the focus of your lab group's research?
The Prober lab focuses on the neural and genetic basis of sleep, using zebrafish as a model. You can learn more about our approach and see our recent publications at The Prober lab website.
What inspires you to aim for sustainable laboratory work?
We are biologists who see the environmental impact of our daily lives and our scientific practices. We feel a social responsibility for our impact on the planet. Our work can sometimes require non-environmentally friendly choices, such as the use of disposable gloves or hazardous chemicals. When our lab makes an effort to reduce the negative effects of our research on the environment with a more sustainable choice, our goals also include possibly improving our technical results, potentially saving funding, and always advancing science while protecting the very thing we study: life on Earth.
We are also hopeful that as we make environmentally friendly choices, other lab groups are encouraged to find creative solutions to make their labs more sustainable as well.
Can you share a specific group undertaking?
Our lab has been able to substitute chemicals we routinely use that are potentially hazardous either to us or to the environment with a green solvent.
What characterizes a green solvent?
A green solvent is a solvent that minimizes the environmental or health impacts of that solvent’s use and disposal.
In what methods has your lab been able to substitute green solvents?
Our lab uses a chemical called formamide, which is toxic to humans and hard to dispose of safely, during a process called in situ hybridization, which is a way to temporally and spatially detect molecules of protein or DNA in a piece of tissue (such as the brain).
What prompted this solvent switch?
Since we became a green-certified lab, our lab members have been inspired to look into greener practices. This particular protocol was being discussed during a lab meeting, and one of our members wondered if there was a safer alternative to formamide. We found several papers* that indicate that ethylene carbonate produces as good as, or better, results for in situ hybridization. Further, upon seeing that we could use ethylene carbonate exactly the same way as we use formamide, which reduced potentially disruptive changes to our protocols, we also found that it is cheaper (about 1/3 of the cost), more environmentally friendly, and healthier for us to use. When we attempted to run an experiment with ethylene carbonate in our lab, we did not observe significant changes to our results. This convinced our lab to make the switch, which was also approved by our EHS.
Prober Lab member (Andrey Andreev) adjusting a two-photon microscope. Photo credit Shannon M West Photography.
Have there been any performance advantages to switching?
By switching to ethylene carbonate, we have not seen any change in the quality of our results, although the papers mentioned suggest their results are improved by using ethylene carbonate. We have only recently made the switch, so have not quantified the data adequately yet to make any broad claims.
There are no changes to the time or process with our FISH protocol which made it easier for our lab to adjust to the change. Adjusting our protocols is not necessarily a barrier to changes, but if we can reduce stress in our lab by making switches as seamless as possible, it’s a big help to convince our members to switch.
The lessened health risks and cost savings are definitely of interest to our lab. We only purchase this chemical about once a year, but over a long period of time and if we find other alternatives to chemicals we commonly use, the monetary, environmental, and health savings will add up.
Could you recommend any resources for other scientists who are interested in exploring green chemistry?
During our research into ethylene carbonate, we found a wonderful website with lots of information on green lab practices, including green chemistry. This site has allowed us to start researching other chemicals that we use in our lab that are hazardous, to see if there are alternatives available for our research. A few other links that we found helpful and inspiring in general include the EPA web page on green chemistry, the Labconscious green lab groups page, and The Wire article Science has a garbage problem what aren’t recycling schemes more popular?
Prober Lab member (Axel Dominguez) filling zebrafish tanks with fresh salt water for putting away recently mated zebrafish (in the mating tanks behind him). Photo credit Shannon M West Photography.
What does it take to convince scientists to replace conventional with a green chemistry alternative?
One thing that our lab struggles with is simply convincing lab members that the change will be beneficial. People are sometimes very attached to particular protocols and resist changing because “what we have works” or “this is the way it’s always been done”. Part of my job in convincing our scientists to change includes testing out the changes, and providing data that in our hands, in our lab, these changes will work.
Further, I show what kind of disruption the changes will cause, and also research the health, environmental, and financial benefits (if any – sometimes changes are more costly or will be harder to procure, and those are certainly barriers). One example of this is changing from Ethidium Bromide (used for staining DNA on a gel to determine genotypes of our animals) to a safer alternative, like Sybr Green, which is more expensive (more than 10x as expensive) but also has many health benefits). Convincing scientists to change is not always easy. Some lab members will resist the changes no matter what kind of data is provided. In turn, some changes are half-way changes, in that new lab members are using the “updated” protocols, and older members are still using the “outdated” protocols.
Are there any other advantages to greening your protocols?
Even though the ethylene carbonate substitution is a small change, making a lot of small changes will make a big impact in our lab. These changes may also encourage other labs to start looking into greener alternatives. It could encourage innovation, thereby making our science and the science in other labs even better. Being a greener lab also increases our visibility. It makes us more attractive to potential collaborators and new students who want to work towards a more sustainable future.
Prober Lab members (Left: Alex Mack, Right: Tasha Cammidge) choosing some zebrafish females to set up for mating. Photo credit Shannon M West Photography.
What do you see as the biggest hurdle for greening biology labs?
Sustainability has not been universally standardized into the operational practices at research institutions, including Caltech. Few lab sustainability practices have support from institutions by way of encouraging integration into operational standards, funding requests, and research practices. (Although some campuses have already adopted these, they are few and far between). As such, many eco-friendly initiatives fail due to a lack of funding.
However, we are hopeful that as more labs on Caltech campus and across the U.S.A. and world adopt changes, such as green chemistry, recycling, composting, green lab certification, etc, that funding opportunities will become more common and integrated into lab facilities management.
References:
Sinigaglia C. et al. A safer, urea-based in situ hybridization method improves detection of gene expression in diverse animal species (2017) Developmental Biology.
Golczyk H. A simple non-toxic ethylene carbonate fluorescence in situ hybridization (EC-FISH) for simultaneous detection of repetitive DNA sequences and fluorescent bands in plants.(2019) Protoplasma.
Kalinka A.et al. Comparison of ethylene carbonate and formamide as components of the hybridization mixture in FISH (2021) Genetics and Plant Breeding