The prospect of using super or enhanced resolution techniques and improving resolution within light microscopy is an exciting prospect, but faced with a whole host of acronyms; SIM, STED, PALM, GSDIM, RESOLOFT to name a few, the prospect of actually picking a technique is less exciting. 

Within the Wolfson Bioimaging facility we have a range of techniques to push the resolution limits of light microscopy:

• Stimulated emission depletion microscopy (STED)
• Single molecule localization microscopy (SMLM)
• SoRa spinning disk microscopy
• Leica Lightning & HyVolution
• Deconvolution microscopy via Huygens (WF and Confocal)

Each of these techniques have different strengths and weaknesses and the choice of which one is best for your project will ultimately be led by the sample and the questions you wish to answer, the highest resolution isn’t always the best option…

The table below is designed to provide a quick overview of the techniques available within the facility and how they compare. Judgements in this table are made by how these techniques perform within the facility, with our current hardware and software options.

More details on each of the techniques can be found using the links above, and should be consulted before committing to a particular technique to ensure your project runs as efficiently as possible. As always, the facility team are always available to answer any questions or discuss projects should you need more information or require further advice.

 

	STED	SMLM	SoRa spinning disk	Lightning/ HyVolution	Deconvolution microscopy
What is the highest XY resolution possible?	approx 50nm	approx 15nm	approx 100-120 nm	approx 130-140nm	approx 150-180nm
what is the highest Z resolution possible?	approx 150nm	approx 15nm	approx 350nm	approx 300nm	approx 350nm
Can I use samples I have already prepared for confocal?
Can I do more than 2 colour imaging?		‌
Does it work well at depth? (>50um)
Is it readily compatible with dynamic live cell imaging?
Is it ‘optical super resolution’? (i.e. no reliance on computer reconstruction etc.)

 

Table 1: overview of different super/enhanced resolution techniques within the wolfson bioimaging facility. A red cross indicates ‘no’ or ‘difficult’, aN amber line indicates ‘possible, but not straightforward’ and a green tick indicates ‘yes’