Electron Microscopy

Just some brief updates of my latest muse - electron microscopy and immunogold labelling (of which I started last Thursday). Perhaps the antibody I probed was abundantly available, so it's pretty awesome to see that the results was as expected for the wild type. As much as I was disappointed that none of the mutants I'm currently looking at has different phenotype when compared to wild type, the data is definitely useful to form future questions for my work. Exciting times ahead!

IgL of my protein of interest - yay!

TEM image of my Chlamybaby - pweeety!!

Here's the paper which I mentioned about the techniques used for IgL. Jeremy is an excellent adviser to me when it comes to imaging techniques. I do my own fixing and cell embedding, so it's always great to have Jeremy around in CAIC whenever I go over. Thanks a lot, Jeremy! :)


Hope you guys find this post helpful :) Cheers!

Immunogold Labelling

Today I've got some peace by chilling at the imaging centre to run some immunogold labelling (IgL). My samples had been fixed in epoxy resin (I'll show you guys some pics in action when I'm running it next week). For now, I'm using older ones, mounted on nickel grids. I'm using a modified protocol from Skepper (2000), J. Microscopy 199, 1-36. (I'll link it later)

I'm having research fun and I think research should be fun. Science is mind blowing and fun. Unfortunately some people have very distorted and disillusioned vision of what "fun" and professionalism in science means.

Right now I'm battling a birdman who wants to kick me out because he's totally disapproving my idea of fun in science. Calling me unprofessional, unproductive and unknowledgeable is low if not bitchy. Telling untruths and causing me this much of troubles at this point only caused me to find him disgusting, and crazy. Stalking me under the disguise of taking my attendance without me knowing is mental harassment.

Maybe some would think that I'm wasting my time battling against a silent but dangerous birdman but if I don't fight until the last drop of my blood runs dry, I'd do myself the biggest harm. As much as I'd like to live in peace to do a PhD, the evil birdman thinks he can show me the door easily. Making my research life difficult is only making the birdman's life worse. I'm a resilient wild grass. Have always been that way.

The birdman said he'd not help with carrying of gas cylinders for me from now on. I'm perfectly happy that he refuses to help me, though I'm totally pissed with his unprofessional and childish behaviour. Thanks to this behaviour I have more conviction that I should fight till the end.

St. Paul mentioned about him running his race till the end. I need to put up a good fight too. If not, I'll be disappointing myself in the end.

Quantitative Western Blotting

I've been recently introduced to infrared fluorescence Western blotting and the video embedded is helpful to understand the differences between IR fluorescence Western blotting and the conventional chemiluminescence method. I'd have to work more on this method before I can be conclusive on its potential and usage but so far, it seems amazing and more cost-, labour- and time-effective. I discovered more from the video of its other functions which I would like to explore further, if given the opportunity to do so.

Two of my samples from the LICOR Odyssey system for the infrared fluorescence Western blot I did this afternoon. 

Does anyone know why there's a red band below the green band when I only use one IRdye?