ThursdayLate!

So we made it to MAG with all our equipment and were ready to set up. We tried a few different layouts before deciding on  one that involved both mine and Nat's artworks in one. The frame worked really well with Nat's large screen prints and I feel they worked fantastically together to raise both pieces form their independent values and produce something really worth while. The collaboration came in the end and I feel that in the hanging stage we were a strong team and really ended up with something special!

I was standing around our area for most of the night and it was great to engage with the audience and explain how and why we had got to this stage. We also had a box with some of the pure culture broth and an agar plate that we got out to show people which went down really well! Overall I think the show was great success and we had great feedback from tutors and visitors alike. 

A pure culture!

Today was the day of the Thursday Late and we were all excited going into the Lab's following the aftermath of yesterdays fantastic results. We took our broths and flowers into the dark room and crossed all of our fingers and toes! There wasn't much happening with the flowers but to our amazement we had successfully harvested a pure culture of bioluminescent bacteria, two of them were really bright but seven out of ten all had a degree of glowing to them. These were the best results yet and I think they were down to the fact that we had looped in the dark and actually been able to see exactly what we were picking up. This was extraordinary and just in time! 

With a few of the less glowing cultures we poured one of them into each of the flowers that we had and decided to make a little hat for the top of the flowers, so they too were in complete darkness, and put them back into the rotating incubator for a few hours longer, before we would need to pick them up to take them over to MAG. 

I also used the wide format printers to print off two images of the glowing bacteria, one on the squid and another of one of three agar plates from the first successful dilutions that we managed to produce. 

The final push

Today was all hands on deck! with some great results from one of our dilution series we made a final attempt at getting a pure culture in the broth, we also had the fantastic idea of looping in the dark so we could actually see the isolated colonies and have a much better shot at looping the right bacteria. We shut all the blinds and turned off all the lights so it was as dark as possible in the space, you need a bunsen burner to sterilise the loop before looping and so the only thing visible was a flame. 

Some of our broth solutions were looking good in the dark so we took some of the best ones and took the swabs out before placing them in the centrifuge. They were in there for 10 minutes rotating at 3800 times a minute so when they came out we were left with all the bacteria forming a pellet at the bottom of the container. After this we drained the excess broth away and added a tiny amount of fresh broth in the hope that the glow would be more concentrated than before. 

We also got one of the squid and put him into a Kilner jar, to see what it would look like, as this was how we were going to display the squid in the exhibition. Filling the jar with salt water would be fine as it was only a short amount of time we needed the squid to be on show for, it didn't matter about preserving it. I really liked the look of the squid pressed up against the jar and the water being slightly foggy just added to the mystery shrouding the squid. I took the jar into the dark room to see if you could still make it out and to my surprise the water had a faint glow to it! When we shook the jar the bioluminescent bacteria seemed to just wash off the squid and make the water in the jar glow.

We then decided to use some of this water to feed the flowers to see if they could take it up before the show tomorrow. we cut the flowers to length and a spliced the bottom of the stems to try and allow easier access into the flowers. filling the flasks with some of the squid water we then bunged the ends of them so there would be no spillage and tin foiled them so that the broth was in darkness, but leaving the flowers able to grow in the light. They then went into the rotating incubator, leaving them for 24hours. 

The last squid

Getting a decent amount of glow on yesterdays squid we got to work, doing two separate dilutions from different squid and taking a lot of swabs from all three to submerge in the broth solutions. We had both Scientists back in the Lab again, as they had completed the majority of their exams now and had a small break before their last one. This was great for morale and also knowledge as they had a much better understanding of the science behind the methods and had completed them a lot more often than I had. As we were nearly running out of agar plates, we settled on using only three dilutions for each series, usually where you would achieve the best results anyway.

As it is the last week, and also the last chance, that we will get to work on any more squid before the exhibition we decided again to get another three squid. These would be the last squid ever that would stink out the microbiology labs, well at least from the Squid Squad anyway!

Tripling our chances

The new start to the week brings new squid, and as it is our last week before the exhibition, we decided to triple our chances and get three fresh squid. While reading up about this experiment we noticed that in one article they mention that it only really works well in 15-20 samples so hopefully this will edge our bets and mean we get some good glowing.

I also gave the plinths their second coat of paint and they are now looking a lot better and more equally covered. We are reaching the end of our time on this project and are trying to get everything to come together for this last push before our Thursday Late event at MAG in a few days!

Friday the13th

Some people are superstitious around this date and believe it brings bad luck, I don't share this view, however it didn't seem to do me any good as none of yesterdays experiments worked! It was all rather discouraging as I had quite enjoyed experimenting the other day and was hopeful that I would yield some good results. Everything had to go into the biohazard waste, as it does every time when we are finished with them, it is very unsatisfying throwing away all of your failed results.  

The day wasn't a complete write off though, I managed to go into the wood workshop and build a few plinths as none of the ones available from MAG were the right size or shape. The wood didn't cost too much and it didn't take very long, I also managed to swipe a bit of paint and gave them their first coat after I got them out of the workshop. As I made them myself I could guarantee that they were both identical, which is really important as the frame in my piece acts as a mirror, so they would need to be the same distance apart and exactly the same shape. 

Squid meets scalpel

I was all alone in the Lab today and after having a chat to one of the technicians the other day I decided that after taking some more swabs to broth solution that I would attempt to slice a thin layer of the skin from where the squid was glowing and submerge it in saline solution to try and subtract some of the more concentrated areas of bacteria. Using an incredibly sharp scalpel and a pair of grippers I cut off three small sections of skin where the glow was best, it felt a bit weird dissecting an animal as it isn't something I have ever done before, but by far the worst part of it was the smell! 

After acquiring the small sections of squid I vortexed the solutions and then, using a pipette, placed 1cl of the solution into a broth solution and 1cl onto an agar plate. I then vortexed the broth solution and wrapped in tin foil ready to go into the rotating incubator and using the glass spreaders again spread out the solution onto the agar plate ready to go into the other incubator. They would be left for another day to allow any growth that might occur. 

I wasn't sure if any of this would produce any good results but it was fun to try something new and also seeing as how our other results seemed so varied it was definitely worth a shot trying any new methods that could work. 

Muck up's and mock up's

Checking back into the lab we discovered that our second dilution series hadn't gone as well as our previous efforts, in fact we had no glowing at all again. This has been such a roller coaster ride of highs and lows, feeling elated when things are going great but also feeling wearied and uninterested when it's not going our way! Since we had no bacteria to sample it was time for squid number seven to make an appearance. 

At the Wednesday session we sorted out our locations for each of the six groups, luckily there was no overlap here, so no need to fight for places. We also had a chat about how are projects were coming along and what we would need for the actual event in just over a weeks time. We discussed the importance of the frame in my piece and I was adamant that it played an important role and that I needed it incorporated into the sculpture. I was advised to test out how it would all look, as I haven't actually seen what it would like, other than in my imagination, although I have a strong vision of what I want to achieve.

Following their advice I decided to go and create a mock up of how I wanted the sculpture to look and how the different aspects would interact together. Using two plinths and hanging an empty frame up in one of the bookable spaces in the studios I played around with different layouts and tried to see how well they complimented each other. I hung the frame at varied heights, swapped between portrait and landscape positions, looked at different distances the plinths were from the frame and even without the frame entirely. I found that I liked the frame hanging portrait and roughly in line with the plinths height, as this allowed you to look through the frame and see both of the objects encased in it. The distance I preferred the further you got from the frame, although this was a smaller space than I would have in the Gallery so I guess I can scope it out there and adjust accordingly. The time spent doing this was highly valuable and only cemented the need for the frame to be a part of my work. 

Dilutions and disasters

Back in the lab we decided to attempt another dilution series seeing as our last one had gone so well. Doing exactly the same procedure as before we took a total of six dilutions before placing them all back in the incubator. We also tried some swabs directly from the squid into broth again and put them into the incubator that keeps them spinning. 

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I checked on the flowers on our windowsill every three hours, taking a photograph to see if there were any alterations. The dark one was taken at 11pm at which point I went to bed and then just checked up again once I had woken up the next morning. 

After a full 24 hours there was still no colour change at all in any of the flowers, the websites I had looked at previously stated that after around six hours there was usually some subtle but noticeable difference. This was a bit of a disappointment and I think I will have to go on the hunt again for some carnations as they are meant to work the best. I had followed the instructions carefully so I expect the problems are down to the type of flowers, these roses did seem to have very thick stems, so maybe that had an impact too. 

Planning & prep

As it is a Monday again we got a new squid from the Arndale fish market, well actually we got two again as last time we had two together it produced the best results we have had so far! We left them up in the lab, same as always, and then left to go to Manchester Art Gallery to have a look around at possible spaces for our Thursday Late, as that is all the work we could complete on the Science side for today. 

We thought we had a scheduled planning and prep meeting at the Gallery, but it turns out we didn't, so we asked if Kate the head curator was around and free, which luckily she was, and managed to have a good discussion with her about our ideas for the evening and how logistics wise it would work. We talked about the security lights being turned off to offer us the darkest possible space, how and where we could hang a frame from, the risk assessments that would need to be completed for bringing in things fro the labs and also about the plinths situation. It was a lot to take in but was a great opportunity to discuss one on one what was achievable and how we could execute it, we even got taken down to the storage spaces downstairs to see if any of the plinths they had there would work for our exhibition. 

This is the space we are wanting to exhibit in as it is the darkest available space which we need so you are able to see the glowing. The bench would obviously be moved and there are lights above that would work for hanging the frame from, the space is quite narrow but I feel the work would fit well into this space. 

I also trailed round a few charity shops looking for some frames to hang for my sculpture, I didn't have much luck but i did find a suitable back up option in Oxfam in Withington. These frames aren't ideal as they are smaller than what I was hoping for but it is nice to know that I have a plan B incase I don't manage to find any others in time. I really want the frames to be as ornate as possible so they really give the impression of a mirror, theses two are alright but I am looking for something a bit grander and more characterful. 

I also tried to get carnations in several different florists around the manchester area, rom Northern Quarter right down to Didsbury, with no luck at all! From my research into which flowers worked best, the close second was roses, I eventually found a bunch of them in Co-op of all places.  

The idea was to test out the dye with the flowers first to see how long they typically take to suck up the dye and change their petal colour, so we would have a rough idea of when the flowers would show if they were glowing or not. I also decided to test out different length of stems and concentration of dye to see if they affected the overall success of the experiment. So I got five glasses and filled them with the same amount of water, some plant food that came with the flowers and different amounts of dye. I pruned them and then cut them to either 15cm, 20cm or 25cm, putting a mixture in each of the different concentrations. It was then time to leave them and see how long it would take them to change colour.

Miniature solar systems

Me and Nat headed into the lab again today to check up on our dilutions to see if we had successfully managed to dilute the bioluminescent bacteria into a more manageable number of colonies so we could more easily pick a single one out. When we looked at our six dilutions it was only the first three that had some glow to them, the last three for whatever reason didn't seem to glow. As you can see in the image below they do get dramatically diluted and maybe it was that there simply weren't enough of the bacteria present for the to glow at all. As a fist attempt at this method and with only the Artists undertaking it due to exams for he BioMed students, I would say we did a pretty good job! (even if I do say so myself)  

When I first came in I forgot my camera so after seeing the amazing results we got I biked back home to pick up my camera and tripod so I could et some nice long exposures of our dilutions. This photo is around a 2 minute exposure, it is only this long because of the low ISO settings, I kept the ISO at 100 so that the image is as clean as possible, with as little noise in it. This photo is also shot on RAW so it will still look great if blown up to a bigger size, which might be an option for our Thursday Late if we can't get any flowers to glow. 

I really like this image, the middle one especially looks to me like a whole solar system with each colony as a star and maybe some of the brighter ones as planets. It is ridiculous to see in person, much better than a photograph can reproduce but it is a good representation for those who come to the Manchester Art Gallery who won't be able to come into the lab to see them. 

Paper circuits and dilutions

I signed up for the EdLab paper circuits workshop a couple of weeks ago and it was on today at 1pm in the Brooks Building, so I missed out on the first talks at the Wednesday meets. The session was really good fun and has lead to talks of helping out for future events with schools. First we looked at simple circuits with LED's and batteries, but after that we had a go at using a CodeBug - a small device that every year 5 and 6 child is given now at school. It is a simple device that easily allows you to complete simple coding on a computer, needing only access to the internet in order to do this. 

The CodeBug plugs into the computer via USB and you can create your own code or download other shared codes from users around the world with a few clicks of a button. I have never had a go at coding before but this simple and easy to learn format meant I could complete a simple message in a relatively quick time frame. Below is a short video of what I managed to do.

In the lab we tried doing a dilution, yet another different method that we have decided to test seeing as we haven't yet got our pure culture. This method involved filling seven small containers with 10cl of saline solution (a mixture of purified water and salt) before directly swabbing the squid on the area where it was glowing brightest. We then broke off the end of the swab in the first container and then vortexed it so the bacteria was completely mixed in to the saline solution. Next we took 1cl of that solution and put it into one of the other unused containers and marked it -1, this was then vortexed and 1cl was taken out and spread on the agar plate with the corresponding number as well as another 1cl being put into the next saline solution -2. This was repeated until we got 6 complete dilutions, each time you were diluting to a tenth so in theory you should get ten times fewer number of bacteria in each solution and on the agar plates. This will hopefully allow us to get some isolated colonies, it should be much easier to pick out an isolated colony if there are only a handful of glowing spots coupled with less growth on the plate overall, which is what we hope will happen here.

The spreaders we used were very different to the swabs and loops we have used previously, they are flat headed so they can spread out the dilution as evenly as possible using the full agar plate. We then put all the agar plates into the shoebox and into the incubator to grow for the next 24 hours. 

After the Wednesday session me and Nat decided to have a chat about where our work was heading as he had come into our group quite late and I have a very fixed idea in my head about what I want the finished piece to look like and how I want it all to come together. I feel like I didn't want to force my ideas onto him and that we both agreed that as well as working on this idea together, he would also pursue some of his own ideas and work separate to ours. I feel like this was a good idea for both of us because as an Artist I want that freedom and I'm sure Nat does too. It was a very much needed discussion and I feel like we both took something from it and can now work better both together and with our independant endeavours.

Squid no. 4

After the exhilaration of last week's successes and in the anticipation of coming back to some great results, we were obviously disappointed to find that none of our batches of broth or agar plates had a single faint glimmer to them. The sad squid below paints a thousand words...

This is the third time in a row now that our trials have failed while left over the weekend. This could be down to a number of different reasons but after some research into our bacteria - vibrio fisheri, we found that it reaches its peak brightness after 24 hours. This means that after this time it starts to get dimmer and dimmer unit there is no glow left at all, it is for this reason that we believe that all our experiments have come to a close over the weekend. As we are not allowed access into the labs for Saturday and Sunday we cannot transfer the bacteria onto new agar plates in time. This also means that time scale is important and that starting with a fresh squid on a Monday grants us the longest time frame in which to continue our experimentation. 

With this we went on our now accustomed route to the arndale to pick up another squid and took it back to the lab, following the, well known by now, procedure and leaving it for another 24 hours. 

Successful submission

Today was all about handing in my essay for my CP unit, due to this I didn't make it into the labs but the others went in my place. The results were good agin, getting further and further with each squid! We didn't have a pure culture but the new agar plates had been successful in transferring some of the bioluminescent bacteria. They used the same loop method and added the bacteria to new broth solutions that went into the incubator again after being wrapped in tin foil.  

Hopes are high again as we leave them over the weekend, although this Monday is a bank holiday so it will be 3 days before we will see any results.

Inoculation and broth

Taking the streaked plates from yesterday into the darkroom we picked out the points with the most isolated cultures and marked them on with a pen. In the light we attempted to loop the bioluminescent bacteria and then put it into 10cl of broth solution, we only scraped the loop along the inside of the container, being careful not to dip it into the solution. Then we used the vortex machine to make sure it was thoroughly mixed in. After we had done two samples form each of the agar plates we then wrapped them up in tin foil, to keep then in darkness, and placed them in the incubator that keeps them at a constant temperature (22°c) and rotates them, so the nutrients in the broth don't just settle at the bottom. 

As well as the broth, we also took a sample from Wednesday's agar plate and inoculated the whole of another agar plate to try and get a whole load of the bioluminescent bacteria on there so we have a bigger selection of colonies to attempt to isolate. Again looking back on previous results and deciding to alter some aspects in order to give us a wider range of possibilities and hopefully produce some more prominent results. Below is a photo of Thursday's efforts (marked in blue) taken from Wednesday's results (the agar plate in the centre). We have put the samples away in their corresponding incubators and will check up on them tomorrow. 

Here is a quick look at the inoculation agar plates, as you can see the swabs are moved across the agar in a cross hatch effect to cover as much of the area as possible to get as much glowing bacteria as we can to grow.

Tony Hall - Tabletop experiments

Today in our wednesday session we were joined by Tony Hall and his big box of goodies! It was a hands on series of experiments, working with batteries, magnets, ink, syringes and even ferrofluid. More than anything it was just fun, taking time out to play around with things reminded me of being a child again. We don't do enough exploring and experimenting but Art and Science are some of the few area's that allow this creative practice to flourish.  

Here is a video of the battery/magnet experiment working, here we just created a completed circuit using wire to connect the battery and the magnet causing the wire to spin around. It is a simple experiment to do, but was fun to reproduce. 

I didn't make it into the lab today as the Science students went in while I was at the tabletop experiments afternoon. They got some great results though, with the broth glowing once it was shaken up. They also got some good glow from the agar plates but again there was too much growth of other bacteria, so they streaked again onto fresh agar, this time trying a slightly altered method. Instead of the usual 4 streaks per section, we went down to 3 and also added another section to try and really spread the vast combination of different bacteria out as thinly as possible to get some isolated colonies. Below is an example of the new improved streaking method. Again they are left in the incubator in the dark and we will return in another 24 hours to check up on them. 

Bioluminescence at it's best!

We got the best glow we have had yet from the two squid together! It was a really big concentration of bioluminescent bacteria from all over the body of the squids. I managed to get a great long exposure in the dark room detailing the full extent of the glowing. With our hopes high we headed into the lab to once again try and isolate the bioluminescent bacteria. 

Using the same technique as before we streaked the agar plates with a swab taken from the part of the squid where the glowing was brightest. Again we took 3 samples from the body, tentacles and water to maximise our chances. 

With the agar plates working but not producing amazing results, we decided it was time to try a new method as well as still attempting to get a pure culture with the old method. So this time we also tried taking a swab and directly submerging it into the broth solution, in the hopes that the broth solution will glow. The container was vortexed using a small machine before being wrapped in tin foil (to stop any light getting in) and then put in the incubator that controls the temperature at the same time as spinning the solution. We need the broth to be in constant motion so that the bacteria doesn't just sit at the bottom and so it can use all the nutrients that are present in the solution. 

We decided to try this new approach after consulting with James, one of the microbiologist researchers, who has worked with bioluminescent algae previously. He suggested taking a direct swab as the glowing was so intense. We will have to wait another 24 hours to see the results!

Third time lucky?

After a long weekend of waiting we were once again greeted by disappointment, there wasn't even a hint of glowing from any of the agar plates. They haven't survived the weekend for whatever reason, it is most probably the time scale rather than the lack of nutrients on the agar plate as we have provided everything they should need. 

So it was off to the arndale centre again to purchase another squid, although this time we got two for no other reason than the minimum card payment was a fiver and we had no change. We took them back to the lab and went through the same procedure as before, covering them in salt water and putting them in the dark in a shoebox, however this time we put the box into the fume cupboard to aid with the smell! At least they have a friend now and might be a little happier together.

Jumping through loops

So after another 24 hours we headed back into the lab and took our agar plates from the incubator and took them into the dark room where we could see if there was any glow. There were quite a few bright spots of light on three of our agar plates, so we tried as best we could to mark where they were (pretty hard in the dark when you couldn't even see the pen you were holding!) hence the messy scribbles.

We then used a teal loop in attempt to isolate some of the bioluminescent bacteria further, as there was a lot of growth from other bacteria and we want a pure culture. This involved heating up the loop to sterilise it before carefully trying to pick up just the glowing bacteria, which you obviously can't see any more, and then re-streaking some fresh agar plates. We use a loop because it is more accurate than a swab, which is what you want when you are trying to get a single colony.

We streaked another three plates from each of the original three so we, in theory, have nine chances of getting an isolated culture. These then go back in the incubator in the dark at 18°c and should be left for another day to grow, although it is Friday now and we don't have access to the labs over the weekend, so we will have to hope that they survive until Monday morning! 

Squid Squad!

We are now known by all the staff in the labs as the Squid Squad, a crack pot of mad scientists/artists with an obsession for deep sea creatures with tentacles! Well we headed back into the lab after roughly 25 hours to check up on our squid and amazingly it had a faint glow to it again - that's a 100% success rate. It wasn't quite as bright as the previous one but there was definitely enough light to proceed to the next step, attempting to isolate the bacteria that we wanted from any others that could be there. To do this we needed some agar plates, swabs, some fashionable blue gloves and most importantly a strong stomach - by now the squid had been rotting in a warm box for over a day and has a certain strong smell about it.

The wonderful staff in the microbiology labs had already made up a number of agar plates and broth with the right concentration of salt and nutrients for our bacteria to thrive. So all we had to do was label and date them correctly and then use the swabs to first get the bacteria from the squid and then wipe it on the agar plates.

There is a proper technique to swabbing the agar plate, used specifically to isolate the bacteria, it is called a streaking technique, I have included a simple diagram explaining the steps to it underneath. We use this certain technique of streaking to try and isolate a single bacteria as there could be lots of different other ones that we don't want. The streaking allows you to drag out the selection of microbes that might be on the swab and get them as thin as possible so some of the bacteria are far away from others and therefore can multiply to form isolated colonies.

We all had a go at this technique, getting two samples from each area - the head, body, tentacles and the water. It was great learning these new skills and fascinating to see how and why you go through all of these procedures. When all the samples were collected we turned all the agar plates upside down, with the solution at the top so that gravity brings down the nutrients in the water after the bacteria have used the top layer. Then we put all the samples into a box to keep them dark again and put them into an incubator at 18°C to keep them at a constant temperature, they need another 24 hours to grow so we will head back tomorrow at around the same time and hopefully see some isolated colonies of bioluminescent bacteria.