What's that smell!

B-mercaptoethanol smells like crap...

Now the lab smells like it... opps...

Phew!

Came in a little late yesterday morning to find an email from the boss...

"Dear SB,
I have a couple of things I would like to talk about.
Are you available for a meeting at say 12 noon today (Monday) ?
Best Regards,
AC"

My instant thought was... "Oh Crap!! I'm in trouble!"

I then started to think of reasons for my poor work output over the past few weeks... (mainly centered around the broken microscope!)

All in vain though - he just wants me to write a review with him... *sigh* more work!

Hey Mum! Look what I made!!

Playing around with GIMP today! GIMP is a FREE image manipulation program... and you can get it at http://www.gimp.org/
I decided to learn about selective colourisation through their tutorial section (www.gimp.org/tutorials/... turned out pretty well!

Backposting...

Please excuse me as I merge journals...

I think I have gone crazy....

as I have already started my thesis... what the?!

Bloody FRETing Red Tape

So more microscope related lab woes... As a lab we are hoping to get into some FRET work. What is FRET? Well, FRET stands for Fluorescent Resonance Energy Transfer and refers to an energy transfer mechanism between two chromophores (or molecules that are responsible for colour. In FRET you have a donor chromophore and an acceptor, when they are in close approximation the donor in its excited state is able to transfer energy to the acceptor (as seen in the figure to the left).

The newly emitted light from the acceptor is different from the donor - leading to a change in the fluorescent spectra which we can measure.

We are hoping to use FRET to see conformational changes in our receptor of interest. By inserting CFP and YFP into different sections of the receptor, we are hoping to measure how FRET differs when receptor is bound to it's various ligands. Do certain sections of the protein move closer or further away from each other to allow ligands to bind? Do they move differently for different ligands? Well... hopefully FRET will allow us to work that out... however for us to do FRET in our lab is requires us to buy new filters for our (probably cursed) miscroscope.

Somehow I managed to be put in charge of buying these filters. Mainly because I require them to try a new FRET based method of measuring intracellular calcium for my STC-1 cells (which don't seem to like Fura-2AM all that much).

So I email "The Company" asking whether our microscope was capable of doing this, what new equipment would we need and (most importantly) how much was it all going to cost. The next day one of "The Company's" reps just happened to be in the building, so they came around, we had a chat and he said that I'd get a quote in the next week.

A week went by....

Two weeks went by... at this stage I'm thinking that our rep has forgotten us so I send off an email. Turns out they had been trying to contact us, but by phone as the rep wanted us to talk to the product manager to make sure we were getting the right filters etc. I ring the product manager while he's in the middle of a conference... but eventually we get in touch, talk about what we need and we finally receive our quote... (with a special 'sorry its taken so long' discount)

The quote offers High Efficiency and Standard Filter Sets... AC (the boss) isn't sure which one we need... it also doesn't list excitation filter holders which we require... so after a few phone calls later we have a revised quote and have (finally) decided what we are getting. Yay! I send an email to 'The Company' and we wait....

After hearing nothing after a week I email our rep who tells me they haven't received an order yet. Puzzled I start to ponder... and then realise that maybe I was meant to put the order through downstairs via our store/purchasing department. (Opps! My Bad!)

The entire order equals up to ~$4,900 containing our filter set (~$4,600) and filter holders (~$300). The next day my one of our postdocs RM comes back to me with the order form. Apparently it had gone over the purchasing department threshold and they couldn't put it through!

So I went down and talked to the store guy who explained that it was over the $5,000 threshold.

I'm Sorry... did I hear that wrong... My order was for less than $5,000... Oh what? Once I add 10% GST ... Ah... yes I see...

So Mr Storeman tells me that I eitehr need to go around and find and cheaper quote (impossible as "The Company" is the only supplier) or get them to write a letter on the company letterhead stating that they are the only supplier for Australia... and hopefully the purchasing department will process it all in about a month or so... (ANOTHER MONTH! ARGGHHH!!)

I decided to have a quick chat to the Boss about it... I was secretly hoping that, as he also doubles as Head of School ,he may have the sway to get this order through without having to try and get this letter... He firsts suggest we split up the order into two, which allows the filter holders to be bought however the filter set is still over once GST is added (by $60!). He also chats to Mr Storeman and brings up the fact that we don't actually have to pay GST on equipment... however it doesn't matter. Letter is still required.

Manage to get into contact with our Rep and explain the situation. After much complaining on the stupidity of the purchasing department by both of us, the rep tells me just to further discount the product so that is = $4,999.00 once GST is added!

Change the price and head down to the store... Explain what I'd done to Mr Storeman, who gives me a very uncertain eye and says "I'll see what I can do."

I haven't heard back yet.... *fingers crossed*

An Article of Substance

With the obligatory "welcome" message out of the way I thought I should probably write a proper journal entry...

At the moment I'm just sort of waiting around the office for our weekly lab meeting to start (in about 2 hours). It's been pretty bludgy for me over the past few week - which is both a good and bad thing. Obviously I would like to be doing more work, however, due to circumstances out of my control (i.e. Our fluorescent microscope being out of order) I've only really been able to do a few other things. I'm seriously hoping for the scope to back and running in the next week or two... so once that happens hopefully it'll be GO GO GO on the experimental front. BUT in all honesty... I'm beginning to think that this microscope is cursed!

First off, our 'waterproof' microscope got liquid all the way through it causing the reflector turret to stop moving automatically. We discovered that it wasn't the turret that was causing the problem... but probably the motherboard. Then the lens off the 63x objective managed to come unglued and could only be replaced by a 40x objective. Then the bulb died on the DG-4 wavelength switcher. When the new bulb was installed, the length of the main connector was changed meaning the light reaching the microscope was more than halved... but we didn't discover that until 3 months of crappy results. Now... it seems as if the bulb of the DG-4 has gone again and since we have no spares we've had to order some in from the US. While this allowed "the company" to take the scope away to fix the turret/motherboard problem... and apparently they manage to break something while transporting it back into the lab. *sighs*

Fingers crossed that this is the last problem we have for a long, long, LONG time.

Hmmm I've just looked out the windows and realised it is looking very dark and gloomy at the moment... not good. Tonight, I am (hopefully) going out for dinner with 'The Girls' (and yes I capitalised that on purpose!), however apparently our booking at the Flying Fajita Sisters is for the outside courtyard and I fear dinner may be cancelled if the weather worsens... which I sincerely hope does not happen as I haven't seen a few of them for a few weeks and I'm really looking forward to a good catch up!

First Post of a New Blog

So throughout my electronic life I've always been a bit of a haphazard blogger/diary writer. I have a tendency to get really enthusiastic and blog 50 million times a day for about 2 weeks... and then it sort of peters out to 6 months of nothing. But... I'm nothing if not a glutton for punishment... so I've decided to start afresh. Move to a new blogging site - try something new. This blog will be a mish mash of personal stuff, uni stuff and anything else that I find interesting.

So if for some reason you've managed to internet wonder you way over here - enjoy :)

Brennan