The next day, we were back in the lab and set about preparing everything we could for when the spin would end and we would be collecting the fish, running our behaviour experiment, and preparing the fish larvae for their return to Bristol for analysis.
We took this time to test our cameras for the behaviour test, label the tubes that the fish were going, and sort out the rest of our equipment to ensure that everything went as smoothly and as quickly as possible when the fish returned to normal gravity. The timed footage from the cameras inside the gondolas was also checked, to see how the larvae were handling the altered gravity. So far, they looked normal, if a bit confused about which way was up!
Before we knew it, evening had come once more and we turned in, wondering what would await us in the centrifuge.
Morning came and our mini-quest resumed. In the lab, Lizzie made up the solutions which would be used to treat the fish after their exposure to hypergravity, while Jessye made sure the camera set ups were functioning correctly. Team Avalanche arrived at lunch, ready to use the centrifuge straight after us for their experiment on avalanches on Mars.
Once the 48 hour timer was up, we donned our lab coats and made a hasty recovery of the petri dishes full of the larvae. Jessye took a dish from each gravity level (6 g, 3 g, 1 g, and 1 g from the lab) of the larvae for the behaviour tests, while Lizzie fixed the remaining fish to take back to Bristol.
The behaviour experiment, as voted for by the public, was simple but effective. The petri dishes were slid into the internally-lit ‘behaviour boxes’ through an opening in the side of the box, and once inside, the mini cameras were started. A minute of ‘normal swimming’ footage was recorded, then the ‘startle test’ began. This involved loudly tapping the outside of the box with a pencil to initiate a startle response from the larval fish from the vibrations. Five startle tests were taken, then the whole process was done again two more times with camera focusing on a different part of the the 15cm diameter petri dish. This entire sequence was then repeated at the following time points: 1.5 hours, 3 hours, and 24 hours after the initial test.
In between the behaviour tests, we looked at the footage from the mini cameras as well as the gondola cameras to see how the larvae had reacted to the hypergravity and how they were readjusting to normal gravity. By the time we had taken our final recording at 24 hours, we could see that the zebrafish larvae that had experienced 6 g initially did not respond to the startle test at all, but after 24 hours had resumed fairly normal swim and startle response behaviour. We compared this to the 1 g lab set of larvae, who all behaved normally from the initial behaviour experiment until the last one.
These initial results seemed to support our hypothesis that hypergravity affects their behaviour, but we will need to run our tracking software on the footage to quantify how different their behaviour was between samples. However, on first appearances, the results are definitely interesting! The effect of hypergravity on the cells and their joint formation are yet to be analysed, but once back in the lab we can stain them and image them to see the impact hypergravity has had on the physiology of the larvae.
We couldn’t wait to get back to the Shire (AKA Bristol) to analyse the larvae fully! But, as all adventuring Hobbits know, tea time is a very important tradition and those poffertjes and stroopwaffles had been at the back of our minds all week… Plus, there was so much of the Netherlands still to explore…
Once we had let wise Gandalf (AKA Chrissy – Lizzie’s supervisor and PI of the Hammond Lab) and the rest of advisers (our Fellowship of the Fish) know that our spin had been a success, we spent the weekend exploring the sights (and foods) of the northern coast.
Too many poffertjes later, we knew it was time to return. We packed our bags, equipment and zebrafish larvae, and headed home to the sunny hills of the Shire. No traffic Balrogs stopped us on our way home, and our travels flew by like a giant eagle.
Lizzie ensured the equipment and larvae were correctly stored in the lab, before spending the next few days catching up on sleep and celebrating with the lab group.
Finally, the legend of the spin was on its path to being fulfilled:
One spin to rule them all, one spin to load them, one spin to bring the fish back and in the dark lab image them.
Bristol Bone Biologists 