Work Experience 2010

From CellBiology

Introduction

Me
My name is Paul and I am a Work Experience student studying in the School of Medical Sciences. My supervisor is Dr Mark Hill, and I have met many people since my first day. This wiki explains in detail what I did over the week that I spent there and how I felt about it.

OHS Induction

Jenny Hartley took me through an OHS Induction on Monday, where I learned about these following things:

  • The potential OHS hazards that could be present in labs and the university as a whole, which could include chemical spills and fires.
  • How to evacuate the building when a fire is present, and who to contact
  • The location of fire exits, fire extinguishers, evacuation assembly points and emergency procedures
  • Location of tea rooms, rest rooms and toilets
  • OHS signs and symbols and their meanings
  • Areas of restricted access
  • How to report a work related hazard

I also had to hand in a form on what I had learned at the OHS Induction by Tuesday.

My Daily Journal

Monday

Bio hood in the tissue culture lab

In the morning, when I first entered the building, Mr Balu Daniel introduced me to Jenny, who went through an OHS induction with me, so that it was safe for me to be in the University.

After that, I was introduced to Dr Mark Hill, who was to be my supervisor for the whole week. He took me to an anatomy practical exam, where students had to go to 10 stations, with 2.5 minutes per station. At each station students were required to answer various questions on the model or diagram displayed. I had to wear a lab coat for the duration of the exam.

Later, I went to observe the same group of students do a Histology and Histopathology exam on the computers. This exam consisted of 6 questions, which each had a 'virtual slide' which had an image of a cell, and students had to identify different components in detail, and answer questions related to the virtual slide. It was extremely boring.

Immediately after this, Dr Hill took me to Jim who is a 4th Year Medical Student. He was in a lab testing whether changes in the pH of the solution in which Retinal Ganglion Cells (RGCS) are placed in affects their survival. The process was very simple. He first got a sheet of parafilm, and divided it into a grid of 5x4 using a marker. He then used a pipette to place 20 microlitres of a liquid on each square. After that, he dipped treated coverslips (which had the retinal cells on them) in Phosphate Buffered Saline (PBS), and placed one on each of the squares on the parafilm. We did this over and over, and he even let me do some!

After a short break for lunch, he continued on with the process from 1pm, while I wrote information on slides for 1 hour till 4pm. After that, I had a chat with Dr Hill about the day's activities, and went to mr Daniel's office and waited for my dad to pick me up from the University.

The cold room in the tissue culture lab

Tuesday

Today Dr Hill came in at 10am, and I went into his office to work on this journal and learned about the different commands. Immediately after Jim came at 11am, I started writing information on the remaining slides while Jim did the same process of dipping cover slips into PBS.

After I finished the slides, Rosh, another student in the medical field, came in and explained in detail the way he was going to conduct his experiment. He was using a method called ELISA. He was researching whether the amount of a certain protein in the spinal fluid was one of the factors for a person to develop Delirium. His method was very simple. Firstly he was going to get a well coated with an antibody called the polyclonal anti-human cystatin C antibody. He then places the Cerebro-Spinal Fluid (CSF - which contains the protein) in the well. He then puts in the same antibody, with an enzyme called Horseradish Peroxidase attached to it. He then adds a substance called substrate into the solution, which produces a precipitate in the bottom of the well. This precipitate turns a blue/black colour and the amount of blue in the precipitate is directly proportional to the protein n the CSF. So then Rosh will make a conclusion on whether the amount of polyclonal anti-human cystatin C antibody in the CSF is one of the factors determining if a person has Delirium.

After that, we took a break for lunch, and Jim had to wait for the cells to incubate, so Rosh and I 'racked tips', which is placing pipette tips on a rack. We did that till 3pm when Rosh had to go to his other supervisor. From then till 4pm I worked on this journal and completed the OHS form I described earlier in this page. At 4pm, Jim came back in, and he started to mount the cover slips onto the slides I wrote information on, and I watched him do that. Half an hour later, my dad picked me up from Jim's lab and we had a chat for 5 minutes. I then went home.

Wednesday

The microscope that Jim used to examine his slides
Today was jam-packed with lots of fun. I met Jim on the bus, so I got straight to work with him. We first went upstairs to a storage room, and we had to wear special lab coats that covered the front of our bodies. Jim cleared out some of his supplies using the needle and a liquid-sucking machine in the bio hood. He showed me the centrifuge, the microscopes, and the -20˚ storage room. It was cold.

After that we examined the slides that we mounted the cells on yesterday under the Inverted Fluorescent Microscope outside Dr. Hill's lab. We had to take pictures from the computer and save them as files as part of Jim's research. We had to get 100 cells for each cover slip. That was hard work trying to find the number of cells required. After a bit of that, we had lunch. We came back later to do some more. It took so long because we had to take 12 pictures per cover slip, and there were 50 slides with 2 cover slips each on them.

Rosh came in later, and I started to help him with writing the letters 'A' and 'B' on little microtubes. It was a hard and lengthy process because the surface on which I had to write on was curved. After that, I went back to help Jim with his slides.

Just after we had finished one batch, Dr. Hill took me to watch a girl called Holly do a practice for her presentation in front of us. I was her 'test', as Dr. Hill said. I had to remember whatever I could from the presentation, and what it was about at the end of it. We then decomposed the whole slideshow and talked about the pros and cons for each slide. I am supposed to see the final presentation on Friday at 1pm.

After that, I came back to the office, and my dad still wasn't there; he was going to be running very late, so I started to work on this journal, and I only went home much later.

Thursday

Today Jim and Rosh weren't at the university, so I was quite disappointed. So I worked on my journal for most of my day.

At 10:30am, I went to the Biomedical Imaging Facility (BMIF), where there were a range of different microscopes, some small, and some really big. A guy called Henry showed me around. You can find more about this at the microscopes section further down the page. After that I worked on my journal for 1 hour, until I took a short break for lunch. After lunch, I worked till 1pm, when I took a walk with Dr Hill to the campus's local Apple Store. There Dr Hill bought Mac OSX Snow Leopard for someone.

At 2:30, we went to watch Holly's improved presentation. It was substantially better. It had slides with good main points, it's layout was much less clutered, and she got rid of the jumbled tables. I had to think up a couple of questions to ask her for after the presentation. She answered them well.

After the presentation, I went back to Dr Hill's office, but he had ti go, so he sent me to Mr Daniel's office to wait and get picked up.

Friday

Today I came in at 10am and worked on my journal till Jim came at 11am. We then went to a quiet room where he examined his cells and saw if the cells were either showing up fluorescent or not. He then had to count the number of cells that were fluorescent. We then both went down to the local apple store, and I got a case for my iPod Touch. I liked it. We then went back, and I fitted the case and screen protector. They fitted well.

After that, we went to the Medical Honours Students' presentations. Holly was up first. This was the 3rd time I had seen her presentation, and it was much better than the previous 2 times put together. It was interesting, had a good layout, and wasn't just text for the whole page. Next was Holly's friend, and she did well, although her slides were overly complicated, and she ended up going overtime. After that there were 2 or 3 more talks that were not so interesting, because they had loads of text and no images. After that I went back to Jim in the quiet room (because he was still examining the cells), and I watched him do that for a bit.

As soon as I got bored of that, I went up to Dr. Hill's room, and worked on this journal on his iPad. It was pretty good, but after a while I gave up and went to the computer to finish of the rest of this journal. I then waited for my dad to come to the university for the last time in a long time to come. I was sad to go, but I knew I had had the best time in my life. The end.

Research Questions

Retinal Ganglion Cells

A Retinal Ganglion Cell (RGCS) is a type of neuron located under the surface of the retina in the human eye. It receives information, in the form of visual information, through two neuron types, bipolar and amacrine cells. RGCS transmit image-forming and non-image-forming information from the retina to various parts of the brain, including the thalamus, hypothalamus, and midbrain. Jim is testing these cells to see whether pH changes affect their survival.

ELISA

ELISA stands for Enzyme-Linked Immunosorbent Assay. It is a type of serological test that is used to test if a certain type of antibody or antigen is present in a cell. This works because every single microbial species has at least one unique antigen. This is divided into 2 sections. You can read more about ELISA here. Rosh is using this technique in his experiment.

High-Tech Microscopes

Early Tuesday morning, I went down to the Bio-Medical Imaging Facility and met Henry who showed me around to a range of interesting microscopes that used cutting-edge technologies.

Axio Observer

The Nuance Microscope

This microscope is encased in a plastic case, or incubated, to maintain the temperature. It uses lasers that allow images to have higher clarity than images that are taken with a microscope using a normal lamp. It can take live and dead cells, which are commonly referred to as 'fixed' cells.

Axio Examiner

This microscope was also incubated to keep the temperature just right, so parts of the microscope didn't heat up or cool down. Both the microscopes that have been mentioned so far are incubated so that they can take live cells, that thrive at body temperature.

Nuance

This isn't incubated, and so, can only take fixed cells, but uses a very expensive camera that uses spectral deconvolution which enables the microscope to view a wide range of colours of colours on the spectrum, which is very important.

The Elyra Microscope

SP5

This microscope can take very high resolution images and can view very deeply into tissue because it uses low frequency infrared rays. It is supported by a smart table that uses air pressure and other technologies to keep the table perfectly still. A smart table is required because the SP5 has an extremely high accuracy, and any movement will reduce the quality of image taking. Another feature of this is that it uses a multi-photon laser, that use extremely powerful light which enables even infrared light to light up dyes, where normal infrared isn't capable of that.

PicoQuant

This is designed to see how proteins move around in membrane; it looks at movement and analyses it, and it can tell whether proteins are fixed in the membrane, or floating around, and if so, how much.

Elyra

The Spinning Disk and TIRF Microscope

This is a prototype that uses the latest technology to make its images many more times accurate than the SP5. Its incubator makes the temperature absolutely perfect. Even the slightest temperature change, even 0.1˚C will make the metal and other objects expand or contract, and make the image inaccurate. It only takes a single image every 30 minutes.

The Spinning Disk And TIRF System

This microscope doesn't have a name as such, but this doesn't make it any less than remarkable. It uses a spinning disk confocal system that allows high speed images to be taken which enables images to be viewed in 3D. It also uses Total Internal Reflection Fluorescent (TIRF) microscopy which does almost the same thing as the PicoQuant, except it is good for surface, rather than deep in the tissue.

You can find more information about the microscopes here

Embryonic Development

Note: all weeks mentioned are clinical weeks, which are 2 weeks more than normal birth weeks due to 2 weeks being before fertilisation.

1st Trimester

  • Week 1: Ovary (Follicle development), Uterine endometrium (loss)
  • Week 2: Ovulation
  • Week 3: Fertilisation
  • Week 4: Implantation
  • Week 5: Neural tube forms
  • Week 6: Heart tube starts beating
  • Week 7: Eyes start forming, nasal pits form, leg and hand buds form
  • Week 8: Brain continues to develop, lungs begin to form, arms and legs lengthened
  • Week 9: Nipples and hair follicles begin to form, elbows and toes visible
  • Week 10: Facial features develop, eyes are more developed
  • Week 11: Tooth buds appear, face is well formed
  • Week 12: Limbs are long and thin, fetus can make a fist with its fingers
  • Week 13: Skin is almost transparent, muscle, tissue and bones have started to develop

2nd Trimester

  • Week 15: Glucagon detectable in fetal plasma
  • Week 19: Nails appear on fingers and toes, eyebrows and eyelashes appear
  • Week 25: 6 alveolar cells type 2 begin to appear and start to secrete surfactant

3rd Trimester

  • Week 31: Body fat rapidly increases, bones fully developed, but soft and pliable
  • Week 35: Fingernails reach the ends of fingertips
  • Week 36: Small breast buds are present, head hair is thick
  • Week 40: Birth

The website that I mainly got my links from was this website ----- http://embryology.med.unsw.edu.au/week/weekbyweek.htm (but I got other little snippets from websites I can't find)