Starting up in science

Part 2: The quest for cash

Alison Twelvetrees leaning over a wired box which makes up part of a microscope

Scientists Alison Twelvetrees and Daniel Bose opened the doors to their new labs at the University of Sheffield, UK, in 2017. 

Their most urgent problem is funding — they can’t keep the lights on without it. 

In June 2018, Ali lost out on a £1-million grant. The year before, the same had happened to Dan.

Can they win funding while trying to design experiments, prepare lectures and supervise students?

This is part two of a three-part series ‘Starting up in science’. Read part one.

5. Team science

Alison Twelvetrees crossing a road in Sheffield carrying a blue ice box

On an overcast morning in October 2018, Ali crosses a busy road clutching an ice box of test tubes. She’s been carrying this precious cargo once a week to the lab of her chief collaborator, biophysicist Tim Craggs, in the University of Sheffield’s physics department.

Their collaboration came about because of a chance encounter before Dan and Ali had even started at Sheffield. On a visit to campus, Dan met Tim and learnt that he had built a special kind of microscope for imaging single molecules. Ali immediately saw the potential for her own work.

“I’m like, ‘Yes. That’s what I need,’” she remembers.

So now she is making the ten-minute walk through the city at a careful pace to take her samples to Tim’s microscope. She is trying to develop a way to study, one by one, the proteins that could form the foundation of her career as a principal investigator (PI). In the depths of the physics building, Ali enters a dimly lit room where a stainless-steel table is kitted out with lasers and lenses. “My son calls it an air-hockey table,” says Tim, gesturing at the experimental set-up. It’s essentially a way to spy on the shapes of the motor proteins called kinesins.

Data on a computer screen attached to a microscope

Data from Ali’s FRET microscopy experiment.

Data from Ali’s FRET microscopy experiment.

Kinesins are shaped like long sticks that have a hinge in the middle and two feet at one end. They are too small to see directly. But Ali has spent weeks creating kinesin proteins adorned with fluorescent tags at each end. The scientists shoot laser light through a diluted drop of cellular fluid from Ali’s test tubes. When an individual kinesin molecule drifts under the laser, its fluorescent tags absorb energy and light up. The frequency of the fluorescence shows whether the motor protein is folded tightly at the hinge or stretched open into its active form. “The actual experiment — don’t tell anyone — is incredibly easy,” says Tim. The harder job, he says, is Ali’s: preparing high-quality protein samples with the right kinds of tags inside cells.

The technique is called fluorescence resonance energy transfer (FRET) microscopy and, much to Ali’s delight, it seems to be working. She hopes to use it to see how single molecules of kinesin change shape when they grab cellular cargo, and then to ask what conditions inside a cell affect that process. This could be the key advance that answers Ali’s burning questions about how cargo travels down neurons at different speeds and what causes this process to go wrong. But it will take months, if not years, to progress from a “quick and dirty” experiment to a repeatable, high-quality protocol, Ali says.

For Ali, the collaboration with Tim and his team, which has been running for about a year, is crucial. It means she has access to a microscope despite having little cash of her own, and other people to bounce ideas off. “When you’re starting out, you’re a team of one. That’s quite a lonely place to be,” she says.

Now, 18 months on, Ali’s lab is a team of three. She’s taken on a PhD student, Evie Smith, who will work full time on the FRET experiment. A final-year undergraduate student, Ashleigh Davey, has also joined the group to work on a ‘neuron-on-a-chip’ project, which will enable the team to watch motor proteins move in cells. Ali has managed to negotiate a move to a larger lab space. But money is still incredibly tight. Evie is funded by a research grant for PhD students, but if Ashleigh is to stay for a PhD, Ali will need to get a grant to support her.

Alison Twelvetrees and colleagues working in a lab near some equipment labelled 'Do not use until trained by Alison Twelvetrees'

Ali's lab, with Ali and Evie in the background.

Ali's lab, with Ali and Evie in the background.

Dan’s team has also grown. Several short-term students have come and gone, but PhD student Petra Celadova is a central figure. Petra joined in November 2017 with years of lab experience already under her belt, and quickly took ownership of her own project even as she helped Dan to set up the lab.

One of her first steps will be to use the gene-editing technology CRISPR–Cas9 to mutate an enhancer in leukaemia cells, looking for those mutations that impair the enhancer RNA’s function. Less than a year into her project, Dan says, Petra is already independent. “She will tell you what to do, which I think is a great sign in a student,” he says. “I take a lot of pleasure when she is like ‘no, this is how we should be doing this’.”

Petra is also wise to the harsh realities of lab work: she hopes to have her cell lines ready by the end of the year, but she knows that research will not necessarily bend to her timetable. “In science, you always think you will have done more than you actually can do in a year,” she says.

Hear Ali and Dan's story in our 4-part podcast miniseries. Via Apple Podcasts | Via Google Podcasts | Via Spotify

6. Second shot

Daniel Bose sitting on some stone steps holding a sheaf of paper as he prepares to go before a panel to get approval for a grant

It’s the eve of Dan’s interview for the prestigious Sir Henry Dale Fellowship awarded by Wellcome and the Royal Society, which he first applied for in 2017 but didn’t get. It’s worth five years of lab funding that he desperately needs. He arrives at a hotel near King’s Cross Station in London just in time to hold a goodnight video call with his three-year-old daughter, Ada.

The next morning, 19 October, Dan wakes up and has an extra-large breakfast to keep his stomach full and his mood positive. As Ada sends him a string of incoherent emoji texts, Dan runs through his practice talk. The interview will take place at the Royal Society’s headquarters near Trafalgar Square, and will consist of a five-minute presentation followed by questions. “It’s quite a lot to get in five minutes,” Dan says. “So I’ve drunk lots of coffee.”

With a bag full of snacks and his nerves jangling, Dan arrives too early at the Royal Society. He strolls through nearby St James’s Park, pausing to watch a group of schoolchildren chase the geese. Then it’s back to the Royal Society, where he sits in the lobby, has some water and tries to gather his thoughts.

There is cause for optimism: the reviews of his proposal were generally positive. He feels confident in the experiments he has proposed. And, in a way, he welcomes the interview experience, however agonizing it might be. The ability to stand up and defend your work is a great skill to have, he thinks. “Do I like it? No. I’m scared,” he says that morning before he leaves for the interview. “But I can see the value of it.”

Daniel Bose walks through the gates of the Royal Society building

Dan walks to his interview at the Royal Society in London.

Dan walks to his interview at the Royal Society in London.

The panel members call him in after their lunch break, and he walks into a room with about 20 people seated in a horseshoe shape. Only two or three seem to be leading the discussion; Dan focuses his eye contact on them. Some of the questions are surprisingly technical, but nothing catches him flat-footed. The 30-minute interview seems like only 10. “I felt like I gave a good impression of myself, and I think that’s all I can do here,” he says. “Other than that, I feel kind of tired.”

Once out of the Royal Society building, his mind returns immediately to his omnipresent to-do list: exam questions to write and all the other things he has pushed to the side while preparing. But first he is going to meet university friends for drinks in a few hours.

He calls Ali, who has spent the day distracting herself at the microscope, and Dan remembers his own anxiety during her interview for the same fellowship four months earlier. He wonders whether the family might go for a walk in the Peak District, a scenic national park near Sheffield, over the weekend to celebrate his new-found liberation from interview prep. “Ada has been bouncing off the walls,” he says. “I wasn’t a very diligent parent for a couple of weeks. I think we might unleash her on some sheep.”

7. Personal best

Daniel Bose sitting in an office reception area smiling

On 1 November 2018, Dan is on a train, sweating over the final details of a presentation he will give that afternoon at University College London. He used to think he would never be the sort of person who would still be preparing their talk on the way to give it. But now he’s alternating between presentation prep and procrastination as he tries to avoid eye contact with his inbox, where a notification from Wellcome could arrive at any moment. “Waiting for e-mails to come in is very anxiety-inducing,” he says.

Temptation eventually gets the better of him and, when he checks, the e-mail is there. “Dear Dr. Bose, I am pleased to inform you …” he reads. Dan’s eyes jump forward to the word “successful” and then he looks away. He glances back and revisits the same words to be sure he hasn’t misread them. “And then I sent it to Ali to double-check and probably read it again,” he recalled later. “But I never made it beyond the first two lines.”

Dan forwards the e-mail to collaborators, his department and people who’d helped with the application. He calls Ali. “We do all of this together. You ride the ups and the downs and the pain and the rejections and all of the stuff you do at the same time,” he says. “So it’s just lovely to be able to share that kind of happiness and release with her.” After a few minutes, the train goes through a tunnel and cuts the call short.

The fellowship will give his lab £1.34 million over 5 years, half of which will be soaked up by salaries for the people who work there. But there will be some money for new equipment. “I’ve been promising Petra here for the last year, all the things we’re going to buy when we’re rich,” says Dan. He thinks about completing her set of micropipettes, buying extra time on the electron microscope and purchasing a machine for fast protein liquid chromatography (FPLC) that purifies proteins. Instead of spending time trying to find ways to stretch his funds to get particular experiments done, the lab could just do them, he thinks. “It just enables everything,” he says. “It makes such a big difference to everything we want to do.”

Lab benches and shelves with little lab equipment
Lab benches and shelves filled with lab equipment and chemicals

Dan’s lab in July 2019

And the lab in September 2021 

The grant also buys Dan time. It funds him for research, guaranteeing some set-aside periods free from the demands of university teaching and committees. Many grants run for only three years; the five-year length of his would allow Dan and Ali to feel more settled. After years of juggling careers, coping with uncertainty and separation, they could look for a school for Ada with the knowledge that she would probably spend much of her primary-school education there. “It’s the first time we’ve been able to say, ‘This is where we’re going to be in five years,’” says Dan. “And that is just making a big difference to everything.”

Would the fellowship affect Dan’s salary? He isn’t sure. “I haven’t really looked at it,” he says. “It’s really very much secondary.”

But for the lab, he has lofty ambitions ready to hand. “Yeah, it’s just going to let us take off as a group,” he says. “That’s what we hope for.”

The next day, Dan kneels down in the middle of the corridor leading to his lab, cradling a bottle of champagne.

“Are we ready?” He shuffles slightly on his knee.

Behind him a gaggle of his colleagues monitor his position on the floor. Their chatter turns to whoops as Dan ejects the bottle’s cork and watches it ping down the corridor and hit the wall just above a poster about a cancer biomarker.

It’s the lab’s tradition to fire off a champagne cork for each new grant; Dan’s last effort, a small award from the Royal Society that began at Easter, was worth £20,000.

Where the cork hits the wall, his colleague affixes a piece of orange tape with Dan’s achievement and the date.

“That’s a PB!” someone shouts.

Orange tape reading ‘Dan B, SHD Fellowship 2/11/18’ on a wall

Dan’s personal best — on tape.

Dan’s personal best — on tape.

Dan having the security of a fellowship “takes the pressure off a little bit”, says Ali. Emotions have been running high as they both digested the news. “Knowing that I didn’t have to worry about Dan any more just kind of came over me and I just felt very overwhelmed.”

The feeling of relief is fleeting. As 2018 draws to a close, Ali is more than halfway through her three-year probation, and time for her to win a grant is fast running out.

In late December, they put those concerns aside to celebrate Christmas with their labs. Last year, Ali and Dan organized a dinner with another new PI, molecular biologist Emma Thomson, who shares lab space with Dan. There were just five diners, including the three lab heads. This year, Ali says, she and Dan had to move a table, chairs and plates to Emma’s house so a dozen people would fit, and “suddenly it was, ‘Ah’, I have built something.’”

8. “I can see myself within that data”

Alison Twelvetrees loading samples onto a microscope in a darkened room

In the new year, the family rents a larger, more comfortable house and instantly feels happier, after two years in what Ali calls an “adequate” rental home. Their kitchen now has a dishwasher, the garden has a swing and room for plants — Ali and Dan start growing Tuscan kale — and the walk to work now takes them through Sheffield’s floral botanic gardens. Ada’s school, which she’ll start this year, is only ten minutes away.

After so many years of moving around, the family feels more settled and able to bring out more of the books and other possessions they’d kept in storage. They make resolutions to throw away what they don’t need. “We’ve accumulated three continents’ worth of possessions,” Dan says.

Alison Twelvetrees sits with her daughter reading a book on the grass in their garden

Ali with the couple’s daughter, Ada, in their garden in Sheffield, UK.

Ali with the couple’s daughter, Ada, in their garden in Sheffield, UK.

At work, Ali has spent much of the past few months setting up protocols and tweaking them over and over again until they work. “It seems so glacially slow sometimes,” says Ali. “But I feel like that’s inevitable, because you’re pulled in so many directions at once. It takes real effort to find the time to do the stuff that you need to do in lab.”

One notable success is that Ali and Emma Thomson have joined together to offer a PhD position split between their labs. Ali’s undergraduate student Ashleigh applied for and won that position, and will start in October 2019 using the neuron-on-a-chip that she had developed to grow single neurons in channels and study ribosomes in the axon. “It’s super, super exciting,” says Ali — but it also puts pressure on her to bring in more research funding.

The Slack messaging group she helped to set up with immunologist Sophie Acton at University College London in 2018 has generated its own workload. Ali and Sophie kicked off a survey of new PIs and their experiences, and by March 2019 they had crunched the data and posted their analysis to the preprint server bioRxiv — since published in the journal eLife (S. E. Acton et aleLife 8, e46827; 2019).

Ali has been diverted by some stark results from the survey, which attracted responses from more than 350 PIs who had launched their first UK labs between 2012 and 2018. Women starting their first research labs tended to have a lower salary and receive less university start-up funding than did their male peers, the results show. Female PIs were also less successful at securing further funding in the first five years.

“I can see myself within that data,” Ali says. Of the two, Dan got a higher salary and more start-up funding — although, Ali adds, they were hired into different positions in different departments, so aren’t directly comparable. “It’s difficult to not compare myself to Dan a lot of the time, because he’s right there. And that’s not particularly healthy,” she says. For Ali, the survey made it clear that the gender pay gap applies to new female lab heads as soon as they start work. “When you’re appointed, you believe that you’re equal to everybody who’s appointed at that stage,” she says. “Actually, it’s not true. We’re being appointed at a deficit immediately.”

Ali takes immediate action on one of the survey findings: that women without mentors reported the lowest level of optimism about their careers. “I could do with more mentorship,” she says. She signs up for a mentorship programme at Sheffield.

Daniel Bose wearing a white coat and gloves pipetting into a centrifuge tube in his lab

Dan pipettes reagents in his lab.

Dan pipettes reagents in his lab.

Dan is finding ways to progress some key experiments while dealing with a burst of teaching, admin and recruitment. He taught a group of students in the lab to clone and purify some of the proteins he’ll need for his cryo-EM experiments. Those students have moved on, but the samples are nearly ready.

He’s also interviewing for new PhD students. With the United Kingdom’s formal exit from the European Union approaching, he’s noticing a drop in applications from Europe. “It’s quite sad. You try to build a culturally diverse laboratory and suddenly no one wants to come because of the uncertainty,” he says.

In late March, Ali decides not to apply for the Sir Henry Dale Fellowship by the rapidly approaching deadline. She is juggling too many priorities to make a strong showing, and is gambling that if she waits until a new round for applications in August, there will be stronger preliminary data from her lab’s work to bolster her bid. But it is a big internal struggle to postpone. If Ali doesn’t win a grant in the next year, she could be out of a job after her university reviews her three-year record. “That will be a disaster for everybody.”

The stress is starting to shake her. “I’m quite up and down. I think I am struggling generally at the moment,” she says when we catch up in April. Starting at Sheffield has not been easy, and before that she felt she was still playing catch-up after maternity leave. “It’s just constantly felt like an uphill battle to try and have any sort of confidence in what I’m doing, and it’s really difficult to kind of be upbeat. Sometimes I just do still feel like I want to quit.”

Alison Twelvetrees sits with her daughter at the edge of a lake by snow covered mountains in Patagonia

The family in Patagonia in 2019.

The family in Patagonia in 2019.

Still, there are jubilant moments. Evie and Ali make their FRET microscope results public for the first time at a conference in Chile in April 2019. “We’re really, really excited about what this method can show us in the future about kinesin activation, although we haven’t discovered any new biology just quite yet,” Ali tells scientists at a small session titled ‘The next generation of cytoskeleton researchers’. Dan flies out with Ada to Chile, so that the family can take advantage of the conference to hike in Patagonia — one of Dan’s bucket-list destinations.

9. Rainbow tape

Different rainbow-coloured tape on a dispense

Dan’s fellowship begins on 1 June 2019, and he quickly sets about laying purchasing plans. He agonizes over what he should buy first, consulting Ali at length. “I had this quandary: you get this grant, what’s the first thing you buy with it?” he says. “It has to be semi-useful but also frivolous. So I got a seven-roll tape dispenser and rainbow tape to put on it.” Then he needs to move on to more-practical purchases, such as centrifuges and shaking incubators. The biggest purchase is the FPLC machine for purifying proteins, with a price tag hovering around £65,000 — more than triple Dan’s budget when he first opened his lab.

Is he at risk of forgetting his humble beginnings? “Maybe ask me in six months’ time, when the gold-plated centrifuge turns up.”

But the clock keeps ticking and, a little more than two weeks after his fellowship began, Dan still hasn’t been able to bring himself to buy the FPLC machine. “Every time I think about it, it brings me out in some kind of horrible sweat,” he says, with a tone that conveys a blend of guilty shopaholic and anxious spendthrift. “It’s kind of terrifying, especially when I’ve been getting by on scavenging the backs of cupboards for a couple of years.”

Petra’s project seems to be going well: she has engineered the different leukaemia cell lines that she will need, and is on the verge of collecting data. Dan’s goal is to get some samples on the cryo-EM over the summer to see how RNAs bind to a protein called CBP that switches genes on and off. Thus far, he has dealt with these molecular complexes only in a test tube. “To actually physically see an interacting RNA and CBP for the first time is going to be super exciting for me,” he says.

Alison Twelvetrees wearing a lab coat and gloves using a pipette in her lab

Ali prepares cell samples.

Ali prepares cell samples.

Ali, meanwhile, is starting to seriously worry about her finances (see ‘Contrasting fortunes’). She’s managed to convert a master’s student at Sheffield, Emma Turner, into a PhD student in her lab, starting in the autumn. For her project, Emma will be engineering mutations into kinesin — the same suite of mutations seen in people with motor neuron disease — to see what effects they have on kinesin walking and unfolding. Her presence will mean that Ali will have doubled the size of her lab team this year, but not increased her income for instruments, reagents and other consumables. “This is my biggest anxiety and stress,” she says.

Contrasting fortunes

Whereas Dan’s large grant allows him to buy new equipment and reagents, Ali has to watch her spending carefully. (Chart excludes staff salaries.)

The students get money to cover their tuition fees and living expenses, but they don’t always get cash for research materials. Evie does have a support grant of about £5,000 a year, which is largely spent on essential equipment such as pipettes and biochemical reagents. Her project can go ahead only because they are using Tim Craggs’s microscope. “I have to be upfront about what we can and can’t afford,” Ali says. She is eyeing a couple of small grants to kickstart and maintain projects for her students.

To top it all, Evie’s microscope experiments, which seemed to be going well, have mysteriously stopped working. “It’s super frustrating,” says Evie. Ali calls it a “summer of anguish”.

By the end of October 2019, as Dan is slowly and anxiously spending his grant money, Ali is midway through her second application for the Sir Henry Dale Fellowship. “Feeling unexpectedly excited/proud of it!” she writes in an e-mail after submitting the full pitch. But this is only step two of four; if they like her application, the judges will interview her in March next year. Then it will be another anxious wait to see if she, too, has secured the money she needs for her lab to survive.

Next: The world changes

Read part 3 of Starting up in science.

Nature 597, 613-618 (2021)


This article is also available as a pdf version.

Authors: Kerri Smith, Heidi Ledford, Richard Van Noorden
Additional Reporting: Benjamin Thompson
Design: Lizzy Brown, Wesley Fernandes, Kelly Krause
Original photography: Chris Maddaloni for Nature
Subeditors: Anne Haggart, Anna Callender
Podcast editors: Benjamin Thompson, Kerri Smith
Editor: Richard Monastersky
Project leader: Kerri Smith

Personal photos provided by Alison Twelvetrees and Daniel Bose.

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