Suggestions regarding Zeiss 5Live

[Christina Pyrgaki]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear all,
We are considering the purchase of a Zeiss 5 Live confocal and we are shortly going to get a demo of the instrument. In our lab we currently use a Zeiss LSM 510 Meta, mainly for the live imaging of mouse embryo development and of mouse lung explants in culture.
I was wondering if people who have used both instruments think that the 5 Live is the instrument of choice when it comes to live imaging applications and is it really so much better compared to the LSM 510 Meta. Also, does anyone have any specific suggestions when it comes to the specifications of the instrument? Is there something that we should look out for if we decide to order the system?
Any suggestion will be greatly appreciated!
Sincerely,

 

Christina Pyrgaki

Niswander Laboratory, HHMI

University of Colorado at Denver

and Health Sciences Center

Department of Pediatrics RC1-N

12800 E. 19th Ave rm 4402E

P.O. Box # 6511

Aurora, CO 80045

TEL. [phone]

FAX. [phone]

 

[K.K.Veeraraghavan]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi

 

Commercial Interest

 

Probably you can also look at Nikon’s Swept Field Confocal too in this class.

 

K.K.Veeraraghavan

Product Specialist

Towa Optics (India) Pvt. Ltd

 

 

 

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christina Pyrgaki
Sent: Thursday, November 22, 2007 11:37 AM
To: [hidden email]
Subject: Suggestions regarding Zeiss 5Live

 

Dear all,
We are considering the purchase of a Zeiss 5 Live confocal and we are shortly going to get a demo of the instrument. In our lab we currently use a Zeiss LSM 510 Meta, mainly for the live imaging of mouse embryo development and of mouse lung explants in culture.
I was wondering if people who have used both instruments think that the 5 Live is the instrument of choice when it comes to live imaging applications and is it really so much better compared to the LSM 510 Meta. Also, does anyone have any specific suggestions when it comes to the specifications of the instrument? Is there something that we should look out for if we decide to order the system?
Any suggestion will be greatly appreciated!
Sincerely,

 

Christina Pyrgaki

Niswander Laboratory, HHMI

University of Colorado at Denver

and Health Sciences Center

Department of Pediatrics RC1-N

12800 E. 19th Ave rm 4402E

P.O. Box # 6511

Aurora, CO 80045

TEL. [phone]

FAX. [phone]

 

[Holly L. AARON]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Christina –

 

Think about why you want to get a 5-Live. Just because it has “live” in the name does not mean it is necessarily better for live cell imaging. In our experience, it bleaches quite quickly and high laser power is often needed. The spinning-disk systems are known to bleach less. Also, some live samples will be happiest with NIR imaging. The 5-Live has some features which make it useful and be sure you do not get the 532nm laser line we are stuck with – it is IMPOSSIBLE to do 2-color simultaneous (green/red) with this laser line. If you go to the far-red (635nm), then it is possible. But let’s face it, most people want to use green/red. But it cannot be done if your laser is right in the middle of your green signal!

 

The best reason IMNSHO to consider the 5-Live is for speed: 2-color simultaneous full-frame 30+ frames per second. The spinning disk systems are still choking on the 2-camera options (as far as I know).

 

I would suggest before plopping down the dough, that you try your sample on a 5live system. That will be the best answer to your question. It is more sensitive than our 510 META system, when it’s aligned well. It is very sensitive to many, many factors – not all of which we have completely figured out yet.

 

Best,

 Holly

__________________

Holly L. Aaron

CRL Molecular Imaging Center

http://imaging.berkeley.edu

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christina Pyrgaki
Sent: Wednesday, November 21, 2007 10:07 PM
To: [hidden email]
Subject: Suggestions regarding Zeiss 5Live

 

Dear all,
We are considering the purchase of a Zeiss 5 Live confocal and we are shortly going to get a demo of the instrument. In our lab we currently use a Zeiss LSM 510 Meta, mainly for the live imaging of mouse embryo development and of mouse lung explants in culture.
I was wondering if people who have used both instruments think that the 5 Live is the instrument of choice when it comes to live imaging applications and is it really so much better compared to the LSM 510 Meta. Also, does anyone have any specific suggestions when it comes to the specifications of the instrument? Is there something that we should look out for if we decide to order the system?
Any suggestion will be greatly appreciated!
Sincerely,

 

Christina Pyrgaki

Niswander Laboratory, HHMI

University of Colorado at Denver

and Health Sciences Center

Department of Pediatrics RC1-N

12800 E. 19th Ave rm 4402E

P.O. Box # 6511

Aurora, CO 80045

TEL. [phone]

FAX. [phone]

 

[Rietdorf, Jens]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Christina & folks,

 

a couple of points I want to add/clarify.

 

>most people want to use green/red

we run our 5 live with 488 & 561 lasers, perfect for this purpose.

 

>spinning disk systems are still choking on the 2-camera options...

we run a dual EMCCD spinning disk since one year.

 

>“live” in the name does not mean it is necessarily better for live cell imaging.

I agree, we mostly use it to scan large mosaics of fixed brain tissue. Here it outcompetes the single beam confocals by at least one order of magnitude in terms of speed, which is remarkable. Scans that used to take over night can now be done within one hour.

 

One of the major issuesto think about is how to handle and store the data; its up to 10GByte per hour, so you can become very popular by filling the entire harddrive and server capacity of your group within one week.

 

regards, jens

 

---

Dr. Jens Rietdorf[hidden email]
Head Microscopy
Novartis Research Foundation
Friedrich-Miescher-Institute, wro1066.2.32
Maulbeerstr.66, CH-4058 Basel, Switzerland
phone [phone] mobil [phone]
Email:rietdorf(at)fmi.ch

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Holly Aaron
Sent: Dienstag, 27. November 2007 00:44
To: [hidden email]
Subject: Re: Suggestions regarding Zeiss 5Live

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Christina –

 

Think about why you want to get a 5-Live. Just because it has “live” in the name does not mean it is necessarily better for live cell imaging. In our experience, it bleaches quite quickly and high laser power is often needed. The spinning-disk systems are known to bleach less. Also, some live samples will be happiest with NIR imaging. The 5-Live has some features which make it useful and be sure you do not get the 532nm laser line we are stuck with – it is IMPOSSIBLE to do 2-color simultaneous (green/red) with this laser line. If you go to the far-red (635nm), then it is possible. But let’s face it, most people want to use green/red. But it cannot be done if your laser is right in the middle of your green signal!

 

The best reason IMNSHO to consider the 5-Live is for speed: 2-color simultaneous full-frame 30+ frames per second. The spinning disk systems are still choking on the 2-camera options (as far as I know).

 

I would suggest before plopping down the dough, that you try your sample on a 5live system. That will be the best answer to your question. It is more sensitive than our 510 META system, when it’s aligned well. It is very sensitive to many, many factors – not all of which we have completely figured out yet.

 

Best,

 Holly

__________________

Holly L. Aaron

CRL Molecular Imaging Center

http://imaging.berkeley.edu

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Christina Pyrgaki
Sent: Wednesday, November 21, 2007 10:07 PM
To: [hidden email]
Subject: Suggestions regarding Zeiss 5Live

 

Dear all,
We are considering the purchase of a Zeiss 5 Live confocal and we are shortly going to get a demo of the instrument. In our lab we currently use a Zeiss LSM 510 Meta, mainly for the live imaging of mouse embryo development and of mouse lung explants in culture.
I was wondering if people who have used both instruments think that the 5 Live is the instrument of choice when it comes to live imaging applications and is it really so much better compared to the LSM 510 Meta. Also, does anyone have any specific suggestions when it comes to the specifications of the instrument? Is there something that we should look out for if we decide to order the system?
Any suggestion will be greatly appreciated!
Sincerely,

 

Christina Pyrgaki

Niswander Laboratory, HHMI

University of Colorado at Denver

and Health Sciences Center

Department of Pediatrics RC1-N

12800 E. 19th Ave rm 4402E

P.O. Box # 6511

Aurora, CO 80045

TEL. [phone]

FAX. [phone]

 

[Scott Phillips-2]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

***commercial interest****

The Andor Revolution Spinning Disk system has a two camera solution with a
dichroic on the emission side that allows for two color capture of any two
wavelengths between 400nm and 700nm.  With the iXon 897 (512x512 BI)
camera it is capable of 30 full fps, with the iXon 860 or a smaller FOV
higher speeds can be reached.

Scott Phillips
Andor Technology
Microscopy Sales