Does anyone has experience with STEDYCON from Abberior? It is described as a user friendly and compact STED unit that can be added on to an existing microscope. What is your experience with it? Have you encountered any problems using it? Thanks!
We have a STEDYCON since 2018 in our facility. Occasionally, the dichroics in the laser box and the scanner require some alignments, so I strongly recommend to have it serviced by Abberior once a year or so (I should add that our instrument was formerly a demo device, which had travelled a lot, so it may be a bit more worn out). Apart from that, we had so far just one minor electronic issue (the laser safety LED indicator panel was broken). It’s indeed compact and fairly user friendly.
we have a STEDYcon in the imaging facility since more than 2 years. If you do not need 3D STED I found the STEDYcon a good compromise and price-wise quite nice. You can do two color STED with resolution up to ~50 nm for the far-red dye and 80-100 nm for the orange dye
Usage is intuitive and due to the design no extra alignment is required by the user. The quality of 2D STED is comparable to Abberior expertline system.
For the quality of the image it is also important on which optics you mount it. Some optics are better suited to cover the wavelengths important for STED (red to IR). As a confocal it is quite slow so it does not replace a standard confocal.
our system performs a bit better, reaching 40 nm with some far-red dyes (preferably Abberior STAR Red) and 60 nm or so with some orange dyes (STAR orange performs well in many stains). We use a 100x objective preselected for a good point spread function, regularily check the STED power (with the built-in powermeter) and, if we observe a STED power drop, occasionally re-align the dichroics in the controller/laser box (the latter, however, requires strict laser safety measures, since it has to be done with an open laser box. Unexperiences microscopists should have this alignment done by the Abberior service team).
for the orange we use Alexa594 and it depends on the sample. For very bright structures one can push the STEDlaser higher and probably get the mentioned resolution. I have to say that 60 nm is pretty good, may be we should try some sample with STAR-Orange.
On our system we have some inherent XY chromatic abberation in the optics (objective/tubus-lens/light path combination). Unfortunately , these can’t be solved by realigning the lasers or dichroics or swapping the objective. The overall quality is a convolution of Abberrior STEDYcon and the body you optics it is mounted on.
What I want to say is that @meetball should look at the microscope body and optics too. May be ask Abberior on which systems they achieve the best results.
We have it mounted on a very simple, unmotorized, upright Zeiss Axio Imager.A2. A minimalistic upright frame is, in this case, probably a good choice. We saw the performance of our STEDYCON first while it was still a demo unit travelling through Europe, mounted on a comparable Olympus (a BX53, if I remember correctly). In this configuration, it also came down to 40 nm in the far-red.
Not an expert about the STEDYCON. Generally speaking, the microscope frame/tube lens from major manufacturers is going to have minimal impact on the image. However at a trade-show or for an on-site demo often sales people are using objective lenses that cost more than the microscope body when they are trying to show off resolution. No mention so far in this post of NA or which manufacturer/model of objective lens???
Thank you! I will surely talk to Abberrior regarding the microscope set up.
We use a Zeiss alpha Plan-Apochromat 100x/NA 1.46 Oil. 100x is the recommended standard magnification for the STEDYCON.