Histology by Ruki and Hafiza

Ruki Ibrahim and Hafiza Irshad were supervised by Dr Laura Yates in Imperial’s Faculty of Medicine. Here they share some of the imaging work they did:

In2Science H&E image 1

In2Science MSB Image 2

In2Science Ecadherin Image 3

In2Science BAL Image 4


The effect of pH on arterial dilation and constriction, by Thulase Sivasothy

Thulase undertook a two-week placement in UCL’s Department of Neuroscience, Physiology & Pharmacology, where she set up experiments to look at the effect of pH on arterial dilation and constriction, as well as trying out patch-clamping.

Read about her experience here: Thulase In2ScienceUK Blog (pdf)

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Mind control?!? by Kiran Pillai

Firstly I want to apologise. The majority of my week or even just a whole day was not based on mind control – it was only a small part. However what I did was as interesting as mind control. So at 8.20 Monday morning I arrive at the Royal London Hospital for Integrated Medicine to attend clinic with Dr. Sharma. What happens here is that patients are referred to the consultant neurologist – Dr. Sharma. He then begins to deduce through a number of different clues; mainly through the patients history and is able to draw on his vast knowledge to correctly diagnose the problem. Actually the most interesting case wasn’t a stunning work of intellectual ability. Rather it came from a patient who already had their diagnosis. They’ve been diagnosed with Cerebral Autosomal-Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy, which is bit of a mouthful (understatement much?). To save everyone’s time and mouth its called CADASIL. This disorder is caused by a mutation to a gene on the nineteenth chromosome. The effect of this mutation is far worse than its name. It dramatically increases the risk of having a stroke and also gives you plenty of migraines. The problem wasn’t exactly the risk of stroke – statins and a healthy diet were all helping to reduce the cholesterol levels. Rather it was the moral dilemmas that are so often skirted over. As it’s a genetic disorder the question was now to worry about the younger generation. The patient’s siblings all ready had the ball rolling in getting them tested. But what about the children? Would any one of us want to know we had this condition and could potentially die from it? The real world implications are huge. They’ll never get any insurance, a mortgage or even a bank loan. They’ll never be allowed to live in America and now a whole plethora of activities have been crossed out of their lives. How would children react if they knew they had the condition? Would it make them live their lives differently? It’s times like these when you truly appreciate the role of a doctor. The doctor has just dropped a massive life-changing bomb on the patient’s shoulders – but his work isn’t confined to just diagnosing. He’s already arranged therapy and counseling for the family and wishes to help them in anyway possible. To be a good doctor, you’re always going to have to balance the science with the social aspects of life and disease.

Moving on from that – Dr. Sharma isn’t just a consultant neurologist; he is also a senior researcher at the MRC Unit for Lifelong Health and Ageing at UCL and that’s where I spent the rest of my days after the clinic. His research is focused on the neurodegeneration of the motor system which he investigates using MRI scans, transcranial magnetic stimulation and the 1946 birth cohort. The first two are pretty science related but the last bit – the birth cohort is pretty amazing. It is a study of 5000 people who were all born in one week during March in 1946; they’ve been followed up throughout their 70 odd years alive. It is the biggest and longest running birth cohort study in the world and so much data; correlation and conclusions have come out of it. For example one such paper that was published using the results showed that the earlier a child would start to walk the higher the IQ. In fact for every month earlier the child learns to stand – on average they gain one half of an IQ point. This has further been proven with the complex link in the brain between the motor cortex and the cognitive ability and thus function of the brain (just want to point out – I could walk pretty much instantaneously after being born).

I bet by now you’re wondering where the mind control comes in. In fact I’ve already mentioned it. Transcranial magnetic stimulation. The act of using magnetic fields to stimulate an electric field in the motor cortex of the brain, which then creates impulses that causes the body’s nervous system to pass on electrical signal, which cause the muscles to contract and move. Currently the motions the body can be made to do are limited but the potential is there. It’s amazing to look at the science behind it, I remember learning in my Physics GCSE about Fleming’s left hand rule – which is exactly what the researchers use to stimulate the brain.

Another great thing about my placement was that my supervisor and me were both similar in one important fact. We are both horrendous in a wet lab. From a chemistry lab to the kitchen you can be guaranteed that I will make mistakes with my hands. So for me; working with computers in dry labs was a great experience. I learnt how to do some coding and use programmes that would actually allow me to do computer modelling of the brain. Using diffusion weighted magnetic resonance imaging I produced stunning images of the brain on my laptop. It’s really interesting how this specific type of MRI scanning works. We all know about the amount of the water our brain holds, but using diffusion MRI we can track motion of water in the brain and produce stunning images of the brain. These images were used to track changes in someone’s brain and then analyse the patterns that come out of it. For Dr Sharma it was how the brain changes with neurodegeneration experienced with motor disease.

The whole week was interesting, educational, inspiring and a number of adjectives that I can’t begin to list. Perhaps the best part was everyone I met whether it was Jane who gave me a cool electronic key to get into a Victorian house or Sanjay who helped me with my medical school entry. But ultimately I have to thank Dr Sharma who opened my eye to the potential for research in medicine; and how a background in both allows a person to approach any scientific problem with a unique perspective that has only motivated me more to become a doctor and now a doctor who is involved in furthering the collective scientific knowledge of the entire human race.

Cancer Research at UCL, by Zoya Gokhool

Graft-versus-host disease (GVHD). The heart of my supervisor’s research are these four words, much like a riddle in its entirety, it would be better to say the heart of this research relies in solving this particular riddle. However this puzzle goes beyond the typical, with its involvement of tissue culturing, fluorescent microscopy, something rather brilliant known as FACS, and much much more. I felt like one of the Doctor’s companions, journeying on a mission, but instead of bow ties and a fez, I had lime gloves and a lab coat, and instead of a Tardis, we had a centrifuge- not nearly as big but it does consider time and relative dimensions (just not in space). The beginning of this mission involves understanding what GVHD actually is; the scenario is as so: bone marrow transplantation is an upheld treatment for curing some blood cancers, but use of this treatment is limited by GVHD. This is when the donor’s T cells attack not only the recipient’s mutated cells, but also their healthy cells. My supervisor Clare Bennett, and her team have a particular focus, which are immune cells known as dendritic cells. Dendritic cells have a vital role in activating donor T cells to attack patient’s cells.

How? This one question brings about a collaboration of procedures, advanced technology and talented scientists, which I had the honour to work with. Amongst the many techniques I had a chance to witness, the most detailed of which was the tissue culturing of tumour cells. A minor fraction of which entailed using several pipettes (much fancier than the plastic ones in school) to get rid of the tumour cells’ waste, feed the tumour cells the necessary nutrients in order for them to grow and incubating the tumour cells. One of the highlights of tissue culturing for me was using a centrifuge; up to that point the existence of a centrifuge was only known to me through biology classes and drawings in textbooks. So to see this in actuality was like seeing a real life unicorn, but only better. See, a centrifuge is a superb machine, which applies centrifugal force to its content; it takes into account the different densities of the substances that are contained within this fluid, and separates them into a pellet and a supernatant each time the fluid is spun – things, which a unicorn most certainly cannot do. And what a great coincidence that the five marker in my biology exam was on this particular machine…

Now onto this brilliance known as FACS- fluorescence activated cell sorting is a specialised type of flow cytometry. It specifies a method by which a mixture of biological cells can be separated into different containers based upon the fluorescent features and specific light scattering of each cell, which is determined by the markers and the proteins the cell contains, and how abundant it is in each particular cell. As part of my tissue culture experiment, I analysed B16.F10 cells (a melanoma tumour cell line). I put tumour cells in six different mediums, the first of which was a negative control. The other five contained a media known as CASM; CASM contains a cytokine known as interferon gamma (IFN-g), which stimulates the tumour cells to express components known as MHC (major histocompatibility complex [class II]), which they normally wouldn’t do in order to avoid the immune system. My association with FACS featured tagging MHC class II with fluorescent dyes, and processing each of the six samples of tumour cells and seeing which samples expressed the components MHC class II. The results are as follow:

Zoya Gokhool FACS poster

Beyond the tissue culture, the FACS and even the awesome portal door that you have to spin clockwise in, in order to reach the dark room to carry out fluorescent microscopy, the best part of this work placement has to be the people I got to work with. Despite being involved with research that is fundamental to an inevitable breakthrough in cancer research in the upcoming future, everyone welcomed me with open arms. There was Clare who warmly received my presence into her lab, Alastair who helped me carry out the entire tissue culture process, Anjum and Sophie who offered invaluable advice on applying to medical school and Anna and Lia who openly shared their research with me, and were always there to share great conversations with. The research, technology and experiments I witness were undeniably fascinating. However the people that make this happen everyday are even better; they have further fuelled my aspiration to becoming a medic, as well as inciting a spark in me to pursue cancer research alongside.

Bath University’s Bee Unit

By Lewis Hayward

The aim of my placement was to isolate a yeast (Metschnikowia pulcherrima) that is capable of producing an antibiotic from blackberry bushes found on campus. I collected 3 leaves, 3 fruits and 3 flowers from 5 bushes on Bath University’s Bee Unit. From there I had to compare yeasts growing on those three structures.

Taking cuttings from each leaf, I positioned them in the lid of a petri dish using the surface tension of water as an adhesive (since the petroleum jelly had run out). For each of the 3 leaves per location, one cutting was face up and the other face down. This was designed to compare the yeasts on the upper and lower surface. The idea was that the ballistospores would be fired onto the agar below. Unfortunately the surface tension was not enough to prevent them falling, so all the microorganisms from within the cuttings grew.

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Mass of fruits and flowers were recorded for each location. For each location, the 3 fruits were placed in a tube of tropic soy broth. Flowers were treated the same. All tubes were shaken for 5hrs to dislodge yeast. Yeast solution was diluted and spread on agar. After a 48hr wait, I had to painstakingly count the yeast colonies and calculate an average. Luckily the desired yeast was distinguishable by its pinky/red colour. I was later able to transfer small amounts of yeast from selected colonies to fresh agar. I discovered that 26.6% of all yeast on fruit and 1.5% of all yeast on flower s was Metschnikowia.

I had the chance to make up a PCR solution containing water, primers and pre-prepared nucleotide solution. Once yeast was added to these, they were placed in the PCR machine. Electrophoresis was used to compare lengths of separated/uncoiled DNA strands to a ladder.

To test bacterial inhibition of various strains of Metsch., I took plugs from each and placed them in plates freshly inoculated with bacteria. A strain called ‘I48’ produced the largest zone of inhibition in the 1st batch.

The experience was genuinely great – a tool that allows one to undertake a real, in-context research task as well as providing insight into the working life of a scientist and finding out about others’ careers. It proves that sometimes repetition, contamination, errors and waiting are part of what it means to be a scientist. Absolutely worthwhile!

Part II: Regina’s second week at KCL

My second week at King’s was spent with Dr Matthew Grubb and his team consisting of Elisa Galliano and Adna Dumitriscu which I got to work with closely with. Their project was more focused on neurons and more specifically, the olfactory bulb in mice. They want to see if and how stimuli affects the AIS (Axon Initial Segment) in neurons where action potentials are generated, whether it affects its length and how far along the axon it is. This can be done with a process called patching where you can observe the action potentials and the way the neurons interact with each other.

I wasn’t able to participate as much in this week as most of the things were technical but I was able to watch and help out in small ways, all the while asking questions about their work. I was able to use a confocal microscope to view some of the zebrafish slides after some immunofluorescence.  This works by targeting antigens with a primary antibody and the fluorophore as a secondary antibody to target that primary antibody so we are able to detect the antigens which in this case is the TH enzyme (tyrosine hydroxylase) in the olfactory bulb. The AnkyrinG also found in the AIS is what enables us to see the latter as they’re the ones being labelled.

 Here is a picture of me using a confocal microscope and seeing some dopaminergic cells found in the CNS and shown due to some immunohistochemistry!

Here is a picture of me using a confocal microscope and seeing some dopaminergic cells found in the CNS and shown due to some immunohistochemistry!

It was a real eye-opener to work in this lab, the two weeks felt really short but I was introduced to so many aspects of neuroscience and how life as a scientist would be so I’m really grateful for this opportunity and that I was able to meet such amazing people. Everyone was so passionate about their project and so eager to help me out in introducing their work and making sure I was able to make the most out of it; I could write about 20 blog entries!

Thank you to In2ScienceUK and everyone at King’s!

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