What can I use to test the performance of a Zeiss 880 Airyscan with PALM super-resolution? It is supposed to have 140nm Airyscan resolution and 20nm super-resolution. I'm looking for something I can source independent of the vendor, preferably which is easy to make or durable. I just had GATTAQuant try to sell me 20nm and 40nm test slides which they then admitted do not twinkle so would not work at all for PALM. They do have 140 and 160nm "nano rulers" which would work for Airy. Perhaps negatively stained diatoms would work, or the fine branched ends of insect tracheoles? I don't culture animal cells, so nuclear pores are not an attractive option.
Hello, just an idea: how about lipid vesicles? These should be easy to make in the laboratory conditions. For the labelling: quantum dots twinkle and their dimensions would fit as well, so you could use them alone or label structures with them.
Yes, we've had vesicles on our confocal systems. They tend to jiggle about a bit, so not ideal for the very long exposures of twinkly super-res. I also am hoping to use a standard with known sized features, so I can prove the performance of the instrument is up to snuff. I was thinking maybe diatoms soaked in dye/buffer could give me a negative image which could be compared against SEM of the same species. Hoping someone out there has a solution they are using for me to copy.
GATTAquant has a PAINT slide that will work for localization super resolution (which covers PALM and STORM). The fluorophores diffuse in and out and bind on and off the DNA origami scaffolds which makes them "blink". You have to image in TIRF with 640 nm, but I don't see why this won't work for the Zeiss PALM system. This will tell you the resolution of the system using that particular fluorophore and will be valuable to know if the system is performing to spec. The resolution of PALM when you use photoswitchable dyes will undoubtedly be worse due to the lower photon yields of photoconvertable/switchable proteins compared to the dye in the QATTAquant slide.
Yes, I was going to get GATTAquant but was then told they are optimized to not blink. I then felt they were trying to get my money for something that they knew wouldn't work. Have you used them yourself for PALM/STORM?
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