opinion on dual camera spinning disc systems

[Katalin Schlett]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all,

I'm new to this list and would be very greatful for any info on the comparison of different SD systems. We are going to establish a small CF with a point scanner and a spinning disc confocal. I would need advice regarding the planned dual-camera SD system, which is going to be used mainly for LCI in cell cultures (cell lines and primary neurons) for FRAP, PA and FRET. So far, I have offers from Zeiss (AxioObserver Z1 + Yokogawa CSU-X1 + Zen Blue + Rapp UGA-42-Firefly), Olympus/Visitron (Olympus IX83 + Yokogawa CSU-X1 + VisiView + VisiScope Confocal FRAP) and Nikon/Andor (Nikon Ti-E + Dragonfly + Manual Micropoint), equipped with different sCMOS cameras.

We have hands-on experience with the Zeiss/Rapp system performing FRAP in dendritic spines of cultured neurons, had only a day-long demo with the Visitron FRAP system and an even shorter demo with the Dragonfly lacking the Micropoint system. Unfortunately, we can not wait until all three systems are available for direct testing and comparison as due to technical and financial restrictions, the decision should be made pretty soon.

I have the following concerns - hopefully, some of you can reflect on them:
1) How well is the Visitron system (especially the VisiView software) integrated with the Olympus IX83? The demo was on a Zeiss Z1 and I had no chance to test the hardver-software communication with the IX83.

2) How fast is the switch between the acquisition and bleaching mode with the Visitron system? I was told that it is about 500 ms, but would like to get a reflection on it, as well.

3) The Dragonfly seemed to be an impressive system, however, I have no clue about the usability of their dual pinhole concept and how efficiently the Manual Micropint can be used. Can any one comment on it?

We have a fixed and pretty tight budget which should cover the point scanning confocal, as well, so we have to accept compromises. Nevertheless, I appreciate any remarks or suggestions.

with best wishes,
Katalin

****************************************
Dr. Katalin Schlett
Associate professor
Eotvos University, Institute of Biology
Dept. Physiology and Neurobiology
1117 Budapest, Pazmany Peter s. 1/C
Hungary

email: [hidden email]

[Sylvie Le Guyader]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Katalin

The CSU-X1 is completely obsolete to my opinion. You need a large field of view of the Dragonfly or the CSU-W to get the best out of your sCMOS camera. Ask those who gave an offer with the CSU-X1 if they can change to an sCMOS with CSU-W and even illumination of the field.

The software capacity and computer architecture must be thouroughly tested if you want to avoid years of head banging. Try timelapse every 10 min for 3h, 4 channels, 4x4 tiles in 10 wells of a multiwell plate to get a small idea of how the software and PC can handle this type of data.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Facility Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Katalin Schlett
Sent: den 20 november 2016 15:41
To: [hidden email]
Subject: opinion on dual camera spinning disc systems

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all,

I'm new to this list and would be very greatful for any info on the comparison of different SD systems. We are going to establish a small CF with a point scanner and a spinning disc confocal. I would need advice regarding the planned dual-camera SD system, which is going to be used mainly for LCI in cell cultures (cell lines and primary neurons) for FRAP, PA and FRET. So far, I have offers from Zeiss (AxioObserver Z1 + Yokogawa CSU-X1 + Zen Blue + Rapp UGA-42-Firefly), Olympus/Visitron (Olympus IX83 + Yokogawa CSU-X1 + VisiView + VisiScope Confocal FRAP) and Nikon/Andor (Nikon Ti-E + Dragonfly + Manual Micropoint), equipped with different sCMOS cameras.

We have hands-on experience with the Zeiss/Rapp system performing FRAP in dendritic spines of cultured neurons, had only a day-long demo with the Visitron FRAP system and an even shorter demo with the Dragonfly lacking the Micropoint system. Unfortunately, we can not wait until all three systems are available for direct testing and comparison as due to technical and financial restrictions, the decision should be made pretty soon.

I have the following concerns - hopefully, some of you can reflect on them:
1) How well is the Visitron system (especially the VisiView software) integrated with the Olympus IX83? The demo was on a Zeiss Z1 and I had no chance to test the hardver-software communication with the IX83.

2) How fast is the switch between the acquisition and bleaching mode with the Visitron system? I was told that it is about 500 ms, but would like to get a reflection on it, as well.

3) The Dragonfly seemed to be an impressive system, however, I have no clue about the usability of their dual pinhole concept and how efficiently the Manual Micropint can be used. Can any one comment on it?

We have a fixed and pretty tight budget which should cover the point scanning confocal, as well, so we have to accept compromises. Nevertheless, I appreciate any remarks or suggestions.

with best wishes,
Katalin

****************************************
Dr. Katalin Schlett
Associate professor
Eotvos University, Institute of Biology
Dept. Physiology and Neurobiology
1117 Budapest, Pazmany Peter s. 1/C
Hungary

email: [hidden email]