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Dear Olivier,
I agree with Gabor and Anton, but would like to add that the important point when discussing CSU-X vs. CSU-W is *power density* (both illumination and detection). This is the main reason the CSU-X can be much faster than the CSU-W. Both are very fast on *paper*, but in reality we are of course photon-limited:
The CSU-W field of view, hence area of illumination is 3.3 times larger. Thus, for the same excitation power, it requires around three times higher laser powers. (It is in fact slightly more complex than this because the CSU-W uses a less dense pinhole spacing too, so requires more like four times greater laser power for equivalence).
On a standard CSU-X and CSU-W, a single-mode fibre is expanded to fill the illumination area. This results in a Gaussian illumination profile - a well-known "feature" of CSUs since the original CSU-10.
On a Borealis-modified CSU-X and CSU-W, a multi-mode fibre (among other changes) is used, resulting in higher potential power densities distributed evenly across the entire field of view.
Thus, Borealis removes the power density issue of the excitation pathway on a CSU-W, but the detection pathway remains the limiting factor - around four times longer exposure times will be required for the equivalent sample due to the larger pinhole spacing and larger field of view.
My opinion would be that, unless your major applications are speed (let's quantify that as 50 Hz or higher), then the CSU-W is a more flexible offering. You can see comparisons of CSU illumination homogeneity with and without Borealis on our website - if large fields of view in fluorescence are a major application, then Borealis would be a dramatic improvement.
Remember also that two camera/ wavelength simultaneous is possible on both the CSU-X (additional optics) and CSU-W (optional extra) but, as Gabor and Anton correctly point out, that identical cameras should be used to facilitate acquisition and analysis.
If you do not want / need to do simultaneous two-camera imaging, then a CSU-W with a large-pixel EMCCD and a small pixel sCMOS might be the ultimate in flexibility. Just use the "dual camera" option to switch the light-path from one camera to another.
I hope this is of some help!
James
James Wainwright
Product Support Engineer (Microscopy Systems)
Europe, Middle East & Africa
Tel: +44 (0) 2890 270 873
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Csúcs Gábor
Sent: 07 October 2015 10:20
To: [log in to unmask]
Subject: Re: Spinning Disk Questions
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Dear Oliver,
We have both an X1 and a W1 system (this is our newest one) and we also have an Andromeda system from FEI (former Till photonics).
Overall you summarize very well what are the things to be decided, but on the other hand only you can evaluate the relative importance of the various factors for your experiments. Here are my comments:
- CSU-X1 or W1
In our newest purchase we went for a W1 because (very) high speed was less an issue for us and the W1 offers more flexibility and a larger field of view (we wanted to have sCMOS cameras).
- sCMOS or EMCCD
Unless you have very weak samples, I do not see so much advantage of an EMCCD. The speed difference is real but the new EMCCD-s can also go up to 60 frames/s and this is not so far from the SCMOS (although much less pixels). In practice, there are only a few projects where you need such extreme speed (typically you simply do not have enough photons to use the full camera speed). A further thing you should consider is the pixel size (Nyquist sampling). With an EMCCD (16 micron pixels) even with a 100x objective you will get (assuming no intermediate magnification) a 160 nm pixel size in your image which can mean undersampling for high NA objectives. This is less an issue with the sCMOS (6.45 micron pixels).
- For two color imaging,
If you mean simultaneous two color imaging - the I would take two identical cameras. Otherwise you will have a hard time to match the two images (EMCCD and sCMOS) to each other due to the different pixel size. On the other hand having both camera types on the system gives you more flexibility.
- Borealis system
It is certainly a nice system (but to my knowledge only offered by Andor), but I do not think that it is absolutely necessary. We have a W1/sCMOS system without it and it is working fine.
- What do you think of silicon objectives ?
I think that the Olympus silicon objectives are indeed superior for live sample applications (compared to normal water ones) but at the same time they are (unfortunately) also very expensive.
Hopefully my comments will help you a little bit.
Greetings Gabor
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Olivier Bardot
Sent: Mittwoch, 7. Oktober 2015 10:18
To: [log in to unmask]
Subject: Spinning Disk Questions
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Hello
We are going to buy our first spinning disk for our facility (we already have CLSM).
We want to image live cells or tissues with lower phototoxicity or increased frames rates.
We would use water objectives from 40x to 63x and 100 frames/seconds will largely be enough for our applications.
Some questions arise for the choice :
- CSU-X1 or W1 : I don't know why we should not go for the new W1. Most companies proposed CSU-X1, but from what I can understand on Yokogawa website W1 is better for most applications. Is there any specific reason to use X1 except frame rates and resolution ?
- sCMOS or EMCCD : W1 will use most of the surface sCMOS detector, not the X1. I know that EMCCD will be more sensitive, but sCMOS are better for speed.
- For two color imaging, is it better to have 1 EMCCD and 1 sCMOS or 2 identical sCMOS ?
- Is Borealis system really necessary for sCMOS detectors ?
- What do you think of silicon objectives ? Are they really better than water objectives for live imaging ?
I know that it will depend on our applications. But I would like to be able to understand the proposal from the vendors.
Thanks
Olivier
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