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On Wed, 17 Dec 2014 06:47:02 +0530, sivaram mylavarapu 
<[log in to unmask]> wrote:
>1) Which is the best microscope in the market for these needs? I have used
>a Zeiss spinning disc in the past, and it appears Leica has an instrument
>too - both use Yokogawa spinning discs. Nikon sells their system with an
>Andor spinning disc.

I've used both the Yokagawa CSU-X and the Andor/Spectral Diskovery system. 
Both on a Nikon Ti-E base. They are comparable in their throughput. The Diskovery 
is much more flexible, and we have it with two camera ports with an EMCCD and a 
CMOS. Also, we can perform confocal and TIRF onto the same cameras without 
moving them. The downside is more complexity, larger footprint, and high cost 
(although the price is approximately the same as if you were going to buy Nikon 
TIRF, Yokogawa disk, and lamp/LED epi).

I would highly recommend either of these systems, especially if your lab has the 
expertise to set up TTL triggering. Neither is exactly "turn-key," but you get 
flexibility.

In order to perform the different imaging modes that you mentioned, you'll either 
have to get the Diskovery (which can move the disk out of the way) or simply 
move the camera from the spinning disk port to another port on the microscope. 
For the latter option, you'll need a separate excitation source, like a lamp or LED 
epi illuminator (I love the Lumencor Spectra-X, but there are many good options). 
If you need laser illumination, such as for TIRF, you'll need a laser combiner that 
has a second fiber output.

>2) Which is the best EMCCD camera for such a system?

I've used Andor iXons (both the 512 and the 1024 pixel versions) as well as CMOS 
(Hamamatsu Flash4 and Andor Zyla 4.2).

I would recommend a 512 EMCCD for spinning disk and single-molecule imaging. 
The flexibility and sensitivity of an EMCCD just isn't yet matched with CMOS. And 
the new EMCCDs run at very fast frame rates. That said, CMOS is cheaper, higher 
resolution, as fast fast fast. I love our CMOS cameras for epi, TIRF, and z-stacks of 
bright samples. For single-molecule, EMCCD is a hair better. (Some of that has to 
do with the larger pixels on the EMCCD. So if your CMOS supports binning, that's 
an option. Also, if you don't mag the detection as much, you can get that dim 
signal into fewer pixels and boost your signal to noise, but then you lose the 
benefit of the high-res camera.)

Most of the companies really use the same sensor, so it matters more that you get 
the best price, that you get good support, and that the camera works well with 
your software. For these reasons, I typically go with Andor, but Photometrics, 
Hamamatsu, etc. are all excellent cameras. (I find that Andor support on Micro-
Manager is superior.)

>3) Can I combine the above features to be supplied with a dual inverted and
>upright microscope so that both high resolution live cell (inverted) and C
>elegans/ Zebrafish/ Drosophila embryo live imaging (upright) can be
>performed as per need?

I'm not 100% sure what you intend to do there. Do you want diSPIM? Nikon and 
other companies are starting to integrate that:
http://www.nikoninstruments.com/About-Nikon/News-Room/US-News/Nikon-
Instruments-Inc.-Unveils-New-Ti-diSPIM-System-at-Cell-Biology-2014

You should be able to add that on top of a spinning disk scope.

I'd be happy to discuss more. Just send me a personal message.

-Sam