What information do I need to bring for discussions of my EM requirements?
- A literature review of published findings and methodologies in the research area
- A brief summary of the main research objectives
- Any relevant preliminary data already obtained.
Do I really need to pay for annual access when I only need to look at a few samples?
No, but you need to discuss this with the Unit; small projects often grow beyond initial expectations meaning that annual access is frequently the most cost-effective option. Short term access usually means engaging in a collaboration or paying £200+ per sample for us to process and image it for you if we have time.
Will someone be available to do all of the sample preparation and imaging for me?
The Unit is a research rather than a service facility meaning that in most cases you will have to provide someone to perform these functions from your group. We will ensure that your staff receive appropriate training and support to achieve your research goals. Alternatively, a number of groups have long-term projects utilising electron microscopy and it may be possible to arrange collaborations with members of these groups
Isn't EM difficult to learn?
Modern TEM's are no more complicated than confocal microscopes and novice users will rapidly reach the level of expertise required to obtain good images of samples. Some preparatory techniques, such as cryo-sectioning, do benefit from long practical experience to obtain the best results; however, with support from EM Unit staff and attention to optimising sample preservation conditions, novice users will invariably obtain results without significant difficulty.
What consumables do I need to get for my EM experiments?
We have all of the chemicals and general consumables required for conventional, immuno- and cryo-EM including EM grids, gold-conjugated secondary antibodies, stains, solvents and resins. For high volume use normally associated with many projects, researchers are expected to replace any consumables used or source their own supplies independently from the Unit.
What do I need to know before attempting immuno-EM?
The technique relies on good antibodies, these should have already been characterised using fluorescence microscopy before considering immuno-EM. Important details include: what dilution does the antibody work at for light microscopy? Typically the antibody will need to be 5-10x more concentrated for EM. Fixation conditions are also critical, ultrastructure can be clearer when glutaradehyde is included during fixation or it may be necessary to use less common fixatives so it is worth testing whether the antibody works under these conditions before progressing to EM. Finally, appropriate negative controls are essential and should also first be verified using light microscopy.
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