ZOYC 2010 suggestions to hosts - Zeiss On Your Campus live cell imaging workshop

[George McNamara]

Dear confocal list,

We just wrapped up a Zeiss On Your Campus live cell imaging workshop
here at UM (www.zeiss.com/zoyc). First, I want to thank all the Zeiss
people involved. This was a huge effort - 20+ boxes (4 of them
crates), two meals, over ten hour X 4 days (plus a good chunk of
Friday) by up to 6 Zeiss folks at any time. Second, I want to thank
everyone in south Florida who attended, and especially the attendees
who made the cuban coffee during the workshop breakfast and lunch.
Third, the people in the DRI building for patience with all the boxes
and cables and attendees. The truckers, and campus electricians, and
other UM staff for helping out. Extra special thanks to Mark
Christenson, Fairchild Imaging, for visiting, speaking and bringing
an sCMOS camera with him (Mark and his camera [5 Megapixels, 33 ms
exposure time] also identified a light leak on our DMIRB optovar -
thanks again Mark, that had been bugging me for a long time but I
never diagnosed the source of the problem with our ORCA-ER).

For those of you who will be ZOYC hosts, I have several suggestions:

* tell your campus security in advance about the truck arrival dates
and approximate time, so they can help stop traffic if needed to get
the truck to your loading dock.

* work with your local Zeiss rep(s) to coordinate the delivery and
pickup. We arranged for ~9am pickup so the Zeiss truck arrived before
the liquid nitrogen/gases and other deliveries. This gave the Zeiss
folks a couple of extra hours on Monday to setup than if the delivery
had had to wait until say noon.

* you need storage space for 20+ boxes for one week (including four
large LSM 780 crates).

* arrange 208 Volt power for the LSM 780 (they have an alternative
using two separate 110 V but our electrician got us 208 V).

* the LSM780 also needed two additional 110 Volt outlets - Zeiss did
not ship extension cords, we had some.

* the LSM 780 came with a vibration isolation table - we supplied a
folding table for the monitor, keyboard, mouse, etc.

* the VivaTome and Observer SD did NOT have vibration isolation
tables (they did come with vistek platforms) - organize some sturdy
tables or free up your scope table(s). We supplied a folding table to
put various VivaTome controller units on.

* Thanks to Zeiss for using 30 inch monitors on all three systems.
Axiovision has a full screen mode (F11). Memo to Zeiss: full screen
mode for ZEN 2010 please, with pan/zoom, etc.

* Zeiss paid for breakfast and lunch for the talks for 70 people
(thanks again Zeiss!) and added fourth talk after lunch by Scott
Olenych on latest fluorescent proteins (thanks Scott!).

* Thanks to Zeiss and Mark Christenson and our UM speakers - we added
several afternoon talks after Scott's post-lunch talk. I was very
pleased by the attendance in the afternoon session.

* Zeiss ships in frozen cells - U2OS osteosarcoma cells with green
and red fluorescence. Have a -80 C freezer and an incubator for them,
and a tissue culture hood to setup in. Also, be happy Zeiss is not
using HeLa cells.

* I encourage buying Sigma-Aldrich M8410 (mouse) embryo tested
mineral oil to use as "liquid coverglasses" to seal imaging dishes
(both Zeiss's and your's). You might also want to have mattek
(glass-bottom-dishes.com), WPI, Greiner Bio-One, EMS, or other 35 mm
imaging dishes on hand for user's cells. Zeiss brought CO2 rigs 9no
tanks) but I decided to go with mineral oil overlays instead of
messing with setting up our CO2 tanks.

* Scott Olenych found that PBS gave better FRAP results on the
Observer SD than whatever culture medium a user brought their cells in.

* Thanks to the Zeiss folks for managing to last through the 4-6pm
BYOS (bring your own specimens) I organized, and for putting up with
several overnight experiments.




If Zeiss ZOYC's UM in 2011, everyone on the listserv is invited to
come over (sorry, you get to pay for your travel). Zeiss will
probably pay for the mini-symposium breakfast and lunch and I will
supply the cuban coffee and find cuban's to make it right. In
addition to what Zeiss brings, we have some nice microscopes, and as
a final inducement, I offer two final words: South Beach.


George McNamara, Ph.D.
Image Core Manager
Analytical Imaging Core Facility
University of Miami, Miller School of Medicine
Miami, FL 33136
[hidden email]
[hidden email]
305-243-8436 office
http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+
spectra .xlsx file is in the zip file)
http://home.earthlink.net/~geomcnamara

[lechristophe]

The Zeiss ZOYC sounds very nice.

Could you or other people on this list elaborate on that sCMOS camera and the technology ?
I found scmos.com but it is mostly marketing.
How does it compare to a hig-end interline CCD (CoolSNAP HQ2 or Orca) and an 1 Mpx EMCCD ?
What would be the price point for such a camera ?

Thanks,

Christophe

On Sun, Mar 21, 2010 at 21:29, George McNamara <[hidden email]> wrote:

Dear confocal list,

We just wrapped up a Zeiss On Your Campus live cell imaging workshop here at UM (www.zeiss.com/zoyc). First, I want to thank all the Zeiss people involved. This was a huge effort - 20+ boxes (4 of them crates), two meals, over ten hour X 4 days (plus a good chunk of Friday) by up to 6 Zeiss folks at any time. Second, I want to thank everyone in south Florida who attended, and especially the attendees who made the cuban coffee during the workshop breakfast and lunch. Third, the people in the DRI building for patience with all the boxes and cables and attendees. The truckers, and campus electricians, and other UM staff for helping out. Extra special thanks to Mark Christenson, Fairchild Imaging, for visiting, speaking and bringing an sCMOS camera with him (Mark and his camera [5 Megapixels, 33 ms exposure time] also identified a light leak on our DMIRB optovar - thanks again Mark, that had been bugging me for a long time but I never diagnosed the source of the problem with our ORCA-ER).

For those of you who will be ZOYC hosts, I have several suggestions:

* tell your campus security in advance about the truck arrival dates and approximate time, so they can help stop traffic if needed to get the truck to your loading dock.

* work with your local Zeiss rep(s) to coordinate the delivery and pickup. We arranged for ~9am pickup so the Zeiss truck arrived before the liquid nitrogen/gases and other deliveries. This gave the Zeiss folks a couple of extra hours on Monday to setup than if the delivery had had to wait until say noon.

* you need storage space for 20+ boxes for one week (including four large LSM 780 crates).

* arrange 208 Volt power for the LSM 780 (they have an alternative using two separate 110 V but our electrician got us 208 V).

* the LSM780 also needed two additional 110 Volt outlets - Zeiss did not ship extension cords, we had some.

* the LSM 780 came with a vibration isolation table - we supplied a folding table for the monitor, keyboard, mouse, etc.

* the VivaTome and Observer SD did NOT have vibration isolation tables (they did come with vistek platforms) - organize some sturdy tables or free up your scope table(s). We supplied a folding table to put various VivaTome controller units on.

* Thanks to Zeiss for using 30 inch monitors on all three systems. Axiovision has a full screen mode (F11). Memo to Zeiss: full screen mode for ZEN 2010 please, with pan/zoom, etc.

* Zeiss paid for breakfast and lunch for the talks for 70 people (thanks again Zeiss!) and added fourth talk after lunch by Scott Olenych on latest fluorescent proteins (thanks Scott!).

* Thanks to Zeiss and Mark Christenson and our UM speakers - we added several afternoon talks after Scott's post-lunch talk. I was very pleased by the attendance in the afternoon session.

* Zeiss ships in frozen cells - U2OS osteosarcoma cells with green and red fluorescence. Have a -80 C freezer and an incubator for them, and a tissue culture hood to setup in. Also, be happy Zeiss is not using HeLa cells.

* I encourage buying Sigma-Aldrich M8410 (mouse) embryo tested mineral oil to use as "liquid coverglasses" to seal imaging dishes (both Zeiss's and your's). You might also want to have mattek (glass-bottom-dishes.com), WPI, Greiner Bio-One, EMS, or other 35 mm imaging dishes on hand for user's cells. Zeiss brought CO2 rigs 9no tanks) but I decided to go with mineral oil overlays instead of messing with setting up our CO2 tanks.

* Scott Olenych found that PBS gave better FRAP results on the Observer SD than whatever culture medium a user brought their cells in.

* Thanks to the Zeiss folks for managing to last through the 4-6pm BYOS (bring your own specimens) I organized, and for putting up with several overnight experiments.




If Zeiss ZOYC's UM in 2011, everyone on the listserv is invited to come over (sorry, you get to pay for your travel). Zeiss will probably pay for the mini-symposium breakfast and lunch and I will supply the cuban coffee and find cuban's to make it right. In addition to what Zeiss brings, we have some nice microscopes, and as a final inducement, I offer two final words: South Beach.


George McNamara, Ph.D.
Image Core Manager
Analytical Imaging Core Facility
University of Miami, Miller School of Medicine
Miami, FL 33136
[hidden email]
[hidden email]
305-243-8436 office
http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file)
http://home.earthlink.net/~geomcnamara

[George McNamara]

Hi Christophe and list,

I have little to add beyond the white paper PDF at http://www.scmos.com/downloads/
I recommend the high resolution pdf version, 8 Mbytes, rather than the 2 Mbytes version .

Seeing the camera in action - the Fairchild engineering is a tiny camera, even with the heat sink. My recollection is the Fairchild camera uses a Camera Link interface ( http://en.wikipedia.org/wiki/Camera_Link), so data transfer is not a problem, and no worries about multiple cameras conflicting, as for example QImaging Firewire cameras.

The data below is from page 9 of the white paper. If there is any data you think is important that I don't mention - well, read the white paper.

The current generation sCMOS is 5.5 Megapixels, front illuminated. Mark mentioned that future developments include a back-illuminated version, which if it works out would narrow the QE advantage of back-illuminated CCDs (peak QE ~90% for standard, Jim Pawley recommends dividing this by 2 when using EMCCD gain) and current sCMOS (peak QE ~60%). 6.5x6.5 um pixel size is about the same as a typical interline CCD (whose peak QE is now about 65% by taking advantage of a microlens and/or transparent gate) but much smaller than most back-illuminated CCD's (usually 16x16 um, some smaller).

In sensitivity and anticipated price (at least for now), the current sCMOS is in between that of a high performance interline CCD and that of back-illuminated CCD or back-illuminated EMCCD. See their figure 12 and 13 for theoretical signal to noise plots (and that the SNR=1 X-axis intercept will move leftwards if a back-illuminated sCMOS comes out).

The key feature - at least in my mind - is that the current sCMOS has over 4x the imaging area of an interline CCD. This means that if it is adapted with the right emission path optics, something like a high efficiency Photometrics QuadView (I wonder if something equivalent to a series of mini-filter cubes be bonded directly to the sCMOS face to produce four channels?), that you would be getting the equivalent of FOUR better-than-interline cameras, for a small price premium over one CCD plus the price of the quad optics (which might be less than the price of a 4-camera splitter unit). The sCMOS also has the ability to read out pixels (or lines or areas) selectively and independently (different exposure times, different rates).

A web search by Mark and I at the ZOYC also turned up a new Hamamatsu Scientific CMOS camera, http://www.microscopy-analysis.com/news/hamamatsu-introduces-new-orca-flash28-scientific-cmos-camera with 2.8 Megapixels (ORCA-Flash2.8), 3 electron readout noise, 45 frames per second full chip, 12-bit Camera link readout. Size of the camera looks about the same as other Hamamatsu cameras. I seem to recall that pixel size is slightly less than 4x4 um.


Enjoy,

George




At 07:14 PM 3/21/2010, Christophe Leterrier wrote:

The Zeiss ZOYC sounds very nice.

Could you or other people on this list elaborate on that sCMOS camera and the technology ?
I found scmos.com but it is mostly marketing.
How does it compare to a hig-end interline CCD (CoolSNAP HQ2 or Orca) and an 1 Mpx EMCCD ?
What would be the price point for such a camera ?

Thanks,

Christophe

On Sun, Mar 21, 2010 at 21:29, George McNamara <[hidden email]> wrote:

Dear confocal list,

We just wrapped up a Zeiss On Your Campus live cell imaging workshop here at UM (www.zeiss.com/zoyc). First, I want to thank all the Zeiss people involved. This was a huge effort - 20+ boxes (4 of them crates), two meals, over ten hour X 4 days (plus a good chunk of Friday) by up to 6 Zeiss folks at any time. Second, I want to thank everyone in south Florida who attended, and especially the attendees who made the cuban coffee during the workshop breakfast and lunch. Third, the people in the DRI building for patience with all the boxes and cables and attendees. The truckers, and campus electricians, and other UM staff for helping out. Extra special thanks to Mark Christenson, Fairchild Imaging, for visiting, speaking and bringing an sCMOS camera with him (Mark and his camera [5 Megapixels, 33 ms exposure time] also identified a light leak on our DMIRB optovar - thanks again Mark, that had been bugging me for a long time but I never diagnosed the source of the problem with our ORCA-ER).

For those of you who will be ZOYC hosts, I have several suggestions:

* tell your campus security in advance about the truck arrival dates and approximate time, so they can help stop traffic if needed to get the truck to your loading dock.

* work with your local Zeiss rep(s) to coordinate the delivery and pickup. We arranged for ~9am pickup so the Zeiss truck arrived before the liquid nitrogen/gases and other deliveries. This gave the Zeiss folks a couple of extra hours on Monday to setup than if the delivery had had to wait until say noon.

* you need storage space for 20+ boxes for one week (including four large LSM 780 crates).

* arrange 208 Volt power for the LSM 780 (they have an alternative using two separate 110 V but our electrician got us 208 V).

* the LSM780 also needed two additional 110 Volt outlets - Zeiss did not ship extension cords, we had some.

* the LSM 780 came with a vibration isolation table - we supplied a folding table for the monitor, keyboard, mouse, etc.

* the VivaTome and Observer SD did NOT have vibration isolation tables (they did come with vistek platforms) - organize some sturdy tables or free up your scope table(s). We supplied a folding table to put various VivaTome controller units on.

* Thanks to Zeiss for using 30 inch monitors on all three systems. Axiovision has a full screen mode (F11). Memo to Zeiss: full screen mode for ZEN 2010 please, with pan/zoom, etc.

* Zeiss paid for breakfast and lunch for the talks for 70 people (thanks again Zeiss!) and added fourth talk after lunch by Scott Olenych on latest fluorescent proteins (thanks Scott!).

* Thanks to Zeiss and Mark Christenson and our UM speakers - we added several afternoon talks after Scott's post-lunch talk. I was very pleased by the attendance in the afternoon session.

* Zeiss ships in frozen cells - U2OS osteosarcoma cells with green and red fluorescence. Have a -80 C freezer and an incubator for them, and a tissue culture hood to setup in. Also, be happy Zeiss is not using HeLa cells.

* I encourage buying Sigma-Aldrich M8410 (mouse) embryo tested mineral oil to use as "liquid coverglasses" to seal imaging dishes (both Zeiss's and your's). You might also want to have mattek (glass-bottom-dishes.com), WPI, Greiner Bio-One, EMS, or other 35 mm imaging dishes on hand for user's cells. Zeiss brought CO2 rigs 9no tanks) but I decided to go with mineral oil overlays instead of messing with setting up our CO2 tanks.

* Scott Olenych found that PBS gave better FRAP results on the Observer SD than whatever culture medium a user brought their cells in.

* Thanks to the Zeiss folks for managing to last through the 4-6pm BYOS (bring your own specimens) I organized, and for putting up with several overnight experiments.

If Zeiss ZOYC's UM in 2011, everyone on the listserv is invited to come over (sorry, you get to pay for your travel). Zeiss will probably pay for the mini-symposium breakfast and lunch and I will supply the cuban coffee and find cuban's to make it right. In addition to what Zeiss brings, we have some nice microscopes, and as a final inducement, I offer two final words: South Beach.

George McNamara, Ph.D.

Image Core Manager

Analytical Imaging Core Facility

University of Miami, Miller School of Medicine

Miami, FL 33136

[hidden email]

[hidden email]

305-243-8436 office

http://www.sylvester.org/AICF (Analytical Imaging Core Facility)

http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file)

http://home.earthlink.net/~geomcnamara

George McNamara, Ph.D.
Image Core Manager
Analytical Imaging Core Facility
University of Miami, Miller School of Medicine
Miami, FL 33136
[hidden email]
[hidden email]
305-243-8436 office
http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file)
http://home.earthlink.net/~geomcnamara