Hi, thank you to Chris for asking me to present today.My name is Tara HardyIll talk to you about my role as a technician now and in my previous roles
I studied Medical Biochemistry at Leicester universityGraduated 2000In my final year I chose to do a laboratory based project which was VacAI really enjoyed the time in the lab, did very well with the project and from this experience I knew that I wanted to work in a lab.
I applied for a technical role in the lab that I did my final year project.
We would receive a bleep if on call, pancreas would arrive in the lab, infused with collagenase, digested, and islets would be seperated out using a gradient, these would then be transplanted into a patient would type 1 diabetes.
Pancreas from a donorIslet transplantationThe islet isolation laboratory that I helped design Distended pancreas using collagenaseThis was only a fixed term contract of a year
Applied for a higher grade technical role making monoclonal antibodies – doing the same as in my final year projectThis also involved …While doing the role I made the uni in excess of £40000
My main role was to make the monoclonal antibodiesTo do this the principal is
When the contract came to an end I decided to try a technical role in industryI applied for a technical role at the Waltham centre for pet nutritionI worked here for 5 years doing a variety of roles, the first being
FISH this is a faeces sample looking at probiotics. Red Lactobacilli that a dog had eaten in the food
I was then a study scientist for a year to cover maternity leave This was not based in the lab, I did miss it
I then went back into the lab but as a lab scientist
Around this time I found out that my father had cancerI made a decision to work in this field because of this but I had to leave waltham and applied for the role I am doing todayI am now a grade 6 lab technician in Biochemistry based here, so back to Leicester university
The imaging facility allow us to see many things we couldn’t years ago. 3D reconstruction can be done as seen here. This is an image of a
These are images of cells that have been fixed and by antibodies we can see the details of the mitosis.
I have been working here three years and have been named author in 4 papers one being …
A good way of doing this is to do a final year project which is lab based
The Pets and People Technician Tara Hardy
Background Studied at The University of Leicester – Medical Biochemistry – Graduated 2000 Final year project - Produced a monoclonal antibody against VacA toxin on Helicobacter pylori From the experience I had at university I knew I wanted to work in a lab.
Grade C Lab Technician University of LeicesterDepartment of Surgery (R. James / S. Swift) Researched Islet cell isolations and Diabetes. Handled human tissue and validating protocols including islet isolation and islet proliferation assays. Part of an on call rota team of six people. Managed my own project researching MMPs in the pancreas. Presented at a Diabetes conference in Austria. Designed an Islet isolation laboratory to a clinical transplantation standard.
Grade D Laboratory Technician University of LeicesterDepartment of Surgery (R. James) Production of monoclonal antibodies. Training BSc and medical students technical assays related to monoclonal antibody production. Tissue culture, Antibody purification, ELISA production, Western blotting, Immunohistochemistry, Bacterialculture, Toxicity assays an animal research. Made Leicester University £40000+ in sales of antibodies. Part of a team that produced an ethical way of monoclonal antibody production. Produced of a web site to advertise monoclonal antibody production at Leicester University.
Laboratory Technician – Waltham Centre For Pet NutritionWaltham on the Wolds, Melton Mowbray
Laboratory Technician – Waltham Centre For Pet Nutrition Part of Masterfoods (Mars Brand). Employ approximately 300 people. Worked in three laboratories, Biochemistry, Tissue culture and Microbiology. Developed assays including the Nitric oxide, cGMP, cortisol and DHEA assays. Haematology analysis, blood gas analysis, radioactive work. Cell proliferation assays, endothelial cell, fibroblast, hepatocyte, and preadipocyte isolation all done in a class 2 contamination laboratory. FISH with DNA and PNA probes.
FISH Technique Cy3 image Overlap of DAPI and Cy3 images
Study Scientist- Waltham Pet centre Designed the trials in this area taking into account ethical requirements. Write protocols for each trial; collate results, data analysis. Liase with all the different teams working at the pet centre. Deal with problems arising and make contingency plans for such occasions Delegation of work and motivating others. Design new ethical techniques to be used in animal oral health in the future. Communication of results in the form of reports, posters and presentations.
Laboratory Scientist – Waltham Pet Centre Wrote protocols and reports for each trial; collate results, data analysis. Liasedwith all the research teams. Dealt with problems arising and make contingency plans for such occasions. Managed/organised five technicians in the microbiology team. Delegated work. Presented at Mars Conferences.
The Comet Assay To assess DNA damage caused to cells by day to day living. The amount of damage can be seen in the extent of the ‘comet’ or tail in the picture
Grade 6 Lab Technician – University of Leicester - Biochemistry Manage the light microscope facilities within the department, responsible for the flow cytometer and Typhoon scanner. Research project (Nek2 substrate – CNap1) Laboratory safety officer and first aider. Supervisor in the Biochemistry teaching program. Being deputy Project License Holder for the Xenopus studies and having responsibility for the mice studies adding mice to my HO Personal License.
Microscopy facilities at Leicester 1. Henry Wellcome Facility 4x Nikon epifluorescence microscopes Leica TCS SP5 confocal laser scanning microscope (CLSM) Eppendorf microinjection facility 2. Adrian Facility Olympus FV1000 CLSM Olympus Cell^R/Scan^R Screening Station Olympus Cytological imaging system 3. MSB Facility Zeiss MP7 Multiphoton CLSM Nikon TIRF microscope Olympus LV200 Bioluminescence imaging system 4. RKCSB Facility Nikon C1Si CLSM
3D reconstructions, maximum intensity projections and z-axis projections
z-series Max intensity projection of 9.5 day old mouse embryo stained for blood vessel network Critchley/Monkley/Spence /Straatman 3D reconstruction of centrosome Prosser/Fry
Immunofluorescence microscopy of fixed cells: observing subcellular localization Centrosomes/Microtubules/DNA
Nature Cell Biology Components of the Hippo pathway cooperate with Nek2 kinase to regulate centrosome disjunction-pp1166 – 1176 Balca R. Mardin, Cornelia Lange, Joanne E. Baxter, Tara Hardy, Sebastian R. Scholz, Andrew M. Fry & ElmarSchiebel The Nek2 kinase directs the separation of duplicated centrosomes. Now the Hippo pathway components hSav1 and Mst2 are shown to regulate the function of Nek2 at the centrosome. This pathway cooperates with the kinesin Eg5 to mediate centrosome separation.
Tips for becoming a Technician Get to know academics with in the university. Make a good impression in your final year. Look for a technical role you are interested in. Be a social person. Be enthusiastic about laboratory work. Want variation in your work. Enjoy Science!