Olympus Cell Excellence TIRF/FRAP

[Rickman, Colin]

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Hi

We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.

Colin

Dr Colin Rickman
Life Science Interface
Department of Chemistry (WP 2.03)
School of Engineering and Physical Sciences
Heriot-Watt University
Edinburgh
EH14 4AS

Tel: [phone] (Office)

http://www.lifescienceinterface.hw.ac.uk
http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

[Chen, De (NIH/NCI) [C]]

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Hi Colin:

We purchased an Olympus TIRF 3 system to do TIRF & PALM imaging. Ours is a customized system, which allows us a lot flexibility in moving parts. The system delivers only 10 % of the laser power to right out of the objective, which is enough for TIRF imaging, but barrly for PALM. Our TIRF lens is 60x, NA 1.49, and a Hammamachu EMCCD camera. We have no big problem to get TIRF images. If you need further information, please let me know.

De

De Chen, Ph.D. (Contractor)  
Scientist I                                                  
Optical Microscopy and Analysis Laboratory
http://atp.ncifcrf.gov                                 
SAIC-Frederick, Inc.                            
National Cancer Institute at Frederick
Post Office Box B – see note below  
Frederick, MD 21702                        
Phone: [phone]
Fax: [phone]
[hidden email]                                      
 
Please consider the environment before printing this e-mail
 
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________________________________________
From: Colin Rickman [[hidden email]]
Sent: Tuesday, May 10, 2011 3:20 PM
To: [hidden email]
Subject: Olympus Cell Excellence TIRF/FRAP

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Hi

We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.

Colin

Dr Colin Rickman
Life Science Interface
Department of Chemistry (WP 2.03)
School of Engineering and Physical Sciences
Heriot-Watt University
Edinburgh
EH14 4AS

Tel: [phone] (Office)

http://www.lifescienceinterface.hw.ac.uk
http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

[Jim Keen]

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Colin,

Regrettably, we have had a very similar experience with this exact system.  We recently returned it to the manufacturer after it could not be made to perform to specs after almost 12 months.  

Feel free to contact me offline for the sordid details.

Jim Keen

James H. Keen, Ph.D.
Professor, Department of Biochemistry and Molecular Biology
Jefferson Medical College
Faculty Supervisor, Bioimaging Facility, Kimmel Cancer Center
Thomas Jefferson University
233 S. 10th St., BLSB/915
Philadelphia, PA 19107

215-503-4624 (Voice)
215-503-0622 (Fax)
[hidden email]
On May 10, 2011, at 3:20 PM, Colin Rickman wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi
>
> We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.
>
> Colin
>
> Dr Colin Rickman
> Life Science Interface
> Department of Chemistry (WP 2.03)
> School of Engineering and Physical Sciences
> Heriot-Watt University
> Edinburgh
> EH14 4AS
>
> Tel: [phone] (Office)
>
> http://www.lifescienceinterface.hw.ac.uk
> http://www.eps.hw.ac.uk/departments/chemistry/cr.htm
>

[Rickman, Colin]

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Hi

When you say the TIRF 3 system do you mean the older CellR system with the triple line condenser controlled by micrometers or the new fully motorized four laser line system? We had the previous microscope system from Olympus which was fine, however, this new motorised system is more problematic.

Colin

Dr Colin Rickman
Life Science Interface
Department of Chemistry (WP 2.03)
School of Engineering and Physical Sciences
Heriot-Watt University
Edinburgh
EH14 4AS

Tel: [phone] (Office)

http://www.lifescienceinterface.hw.ac.uk
http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

On 10 May 2011, at 22:35, Chen, De (NIH/NCI) [C] wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Colin:
>
> We purchased an Olympus TIRF 3 system to do TIRF & PALM imaging. Ours is a customized system, which allows us a lot flexibility in moving parts. The system delivers only 10 % of the laser power to right out of the objective, which is enough for TIRF imaging, but barrly for PALM. Our TIRF lens is 60x, NA 1.49, and a Hammamachu EMCCD camera. We have no big problem to get TIRF images. If you need further information, please let me know.
>
> De
>
> De Chen, Ph.D. (Contractor)  
> Scientist I                                                  
> Optical Microscopy and Analysis Laboratory
> http://atp.ncifcrf.gov                                 
> SAIC-Frederick, Inc.                            
> National Cancer Institute at Frederick
> Post Office Box B – see note below  
> Frederick, MD 21702                        
> Phone: [phone]
> Fax: [phone]
> [hidden email]                                      
>
> Please consider the environment before printing this e-mail
>
> This e-mail and any attachments to it are intended only for the identified recipients.  It may contain proprietary or otherwise legally protected information for SAIC-Frederick.  Any unauthorized use or disclosure of this communication is strictly prohibited.  If you have received this communication in error, please notify the sender and delete or otherwise destroy the e-mail and all attachments immediately.
>
>
> ________________________________________
> From: Colin Rickman [[hidden email]]
> Sent: Tuesday, May 10, 2011 3:20 PM
> To: [hidden email]
> Subject: Olympus Cell Excellence TIRF/FRAP
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi
>
> We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.
>
> Colin
>
> Dr Colin Rickman
> Life Science Interface
> Department of Chemistry (WP 2.03)
> School of Engineering and Physical Sciences
> Heriot-Watt University
> Edinburgh
> EH14 4AS
>
> Tel: [phone] (Office)
>
> http://www.lifescienceinterface.hw.ac.uk
> http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

[Rickman, Colin]

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*****

Hi

Thanks for getting in touch. We are having significant problems withe the TIRF module on this system. The TIRF does work but only if you manually adjust the laser angle in the software. The preset 100 nm TIRF certainly is 100 nm penetration. We currently have four laser lines attached (405 nm, 491 nm, 561 nm, and 640 nm) and the 640 nm seems to be suffering from a large amount of scattering in the light path. All of this is using the 150 x 1.45 NA lens. Members of Olympus OSIS have flown over to look at the system tomorrow (OSIS in Germany is where the system was developed) so hopefully they can resolve some of the issues. they are also struggling to supply any laser safety interlocked system which is compatible with our stage and incubation system. I think that is going to be more difficult to rectify.

What was wrong with your system? Did you buy an alternative system? If so would you mind letting me know which you went for. We are a bit restricted in our choice as we really need to use the high magnification high NA lens available from Olympus.

Colin


Dr Colin Rickman
Life Science Interface
Department of Chemistry (WP 2.03)
School of Engineering and Physical Sciences
Heriot-Watt University
Edinburgh
EH14 4AS

Tel: [phone] (Office)

http://www.lifescienceinterface.hw.ac.uk
http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

On 11 May 2011, at 02:17, Jim Keen wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Colin,
>
> Regrettably, we have had a very similar experience with this exact system.  We recently returned it to the manufacturer after it could not be made to perform to specs after almost 12 months.  
>
> Feel free to contact me offline for the sordid details.
>
> Jim Keen
>
> James H. Keen, Ph.D.
> Professor, Department of Biochemistry and Molecular Biology
> Jefferson Medical College
> Faculty Supervisor, Bioimaging Facility, Kimmel Cancer Center
> Thomas Jefferson University
> 233 S. 10th St., BLSB/915
> Philadelphia, PA 19107
>
> 215-503-4624 (Voice)
> 215-503-0622 (Fax)
> [hidden email]
> On May 10, 2011, at 3:20 PM, Colin Rickman wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi
>>
>> We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.
>>
>> Colin
>>
>> Dr Colin Rickman
>> Life Science Interface
>> Department of Chemistry (WP 2.03)
>> School of Engineering and Physical Sciences
>> Heriot-Watt University
>> Edinburgh
>> EH14 4AS
>>
>> Tel: [phone] (Office)
>>
>> http://www.lifescienceinterface.hw.ac.uk
>> http://www.eps.hw.ac.uk/departments/chemistry/cr.htm
>>

[Chen, De (NIH/NCI) [C]]

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*****

Dear Colin:

You may be right. Our TIRF 3 first came with three laser arms, with TIRF angles of the three laser tuned by micrometers. But we recently motorized the three arms. After motorization, TIRF has to be tuned by software, which is not convenient. The good for motorization is that it allows fast switch between TIRF and non TIRF mode. You problem I guess is that, using software, each step changes the incidence angle by very little, so it becomes difficult for you to fine tune to get the best TIRF illumination. If this is your problem, you should be able to solve it by changing the step size in software, making each click generate more motor move distance. That's what we had done here. I would be happy of this info helps any bit.


De


De Chen, Ph.D. (Contractor)
Scientist I
Optical Microscopy and Analysis Laboratory
http://atp.ncifcrf.gov
SAIC-Frederick, Inc.
National Cancer Institute at Frederick
Post Office Box B – see note below
Frederick, MD 21702
Phone: [phone]
Fax: [phone]
[hidden email]


________________________________________
From: Colin Rickman [[hidden email]]
Sent: Thursday, May 12, 2011 3:44 PM
To: [hidden email]
Subject: Re: Olympus Cell Excellence TIRF/FRAP

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi

When you say the TIRF 3 system do you mean the older CellR system with the triple line condenser controlled by micrometers or the new fully motorized four laser line system? We had the previous microscope system from Olympus which was fine, however, this new motorised system is more problematic.

Colin

Dr Colin Rickman
Life Science Interface
Department of Chemistry (WP 2.03)
School of Engineering and Physical Sciences
Heriot-Watt University
Edinburgh
EH14 4AS

Tel: [phone] (Office)

http://www.lifescienceinterface.hw.ac.uk
http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

On 10 May 2011, at 22:35, Chen, De (NIH/NCI) [C] wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Colin:
>
> We purchased an Olympus TIRF 3 system to do TIRF & PALM imaging. Ours is a customized system, which allows us a lot flexibility in moving parts. The system delivers only 10 % of the laser power to right out of the objective, which is enough for TIRF imaging, but barrly for PALM. Our TIRF lens is 60x, NA 1.49, and a Hammamachu EMCCD camera. We have no big problem to get TIRF images. If you need further information, please let me know.
>
> De
>
> De Chen, Ph.D. (Contractor)
> Scientist I
> Optical Microscopy and Analysis Laboratory
> http://atp.ncifcrf.gov
> SAIC-Frederick, Inc.
> National Cancer Institute at Frederick
> Post Office Box B – see note below
> Frederick, MD 21702
> Phone: [phone]
> Fax: [phone]
> [hidden email]
>
> Please consider the environment before printing this e-mail
>
> This e-mail and any attachments to it are intended only for the identified recipients.  It may contain proprietary or otherwise legally protected information for SAIC-Frederick.  Any unauthorized use or disclosure of this communication is strictly prohibited.  If you have received this communication in error, please notify the sender and delete or otherwise destroy the e-mail and all attachments immediately.
>
>
> ________________________________________
> From: Colin Rickman [[hidden email]]
> Sent: Tuesday, May 10, 2011 3:20 PM
> To: [hidden email]
> Subject: Olympus Cell Excellence TIRF/FRAP
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi
>
> We recently purchased an Olympus Cell Excellence TIRF system with four independent laser lines and the additional hardware FRAP scanner. We are having numerous ongoing problems with both the hardware, software and support from the UK arm of Olympus microscopes. I was wondering if anybody else has taken delivery of such a system (with the new 4 line motorised TIRF combiner) and if they are having problems.
>
> Colin
>
> Dr Colin Rickman
> Life Science Interface
> Department of Chemistry (WP 2.03)
> School of Engineering and Physical Sciences
> Heriot-Watt University
> Edinburgh
> EH14 4AS
>
> Tel: [phone] (Office)
>
> http://www.lifescienceinterface.hw.ac.uk
> http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

[Andreas Bruckbauer]

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Are you sure they have put in a laser line filter for the 640 nm laser? They usally have some spurious emission which away from the main line. This causes a lot of problems.

best wishes

Andreas

 

 


 

 

-----Original Message-----
From: Colin Rickman <[hidden email]>
To: [hidden email]
Sent: Thu, 12 May 2011 20:44
Subject: Re: Olympus Cell Excellence TIRF/FRAP


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To join, leave or search the confocal microscopy listserv, go to:

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Hi



Thanks for getting in touch. We are having significant problems withe the TIRF

module on this system. The TIRF does work but only if you manually adjust the

laser angle in the software. The preset 100 nm TIRF certainly is 100 nm

penetration. We currently have four laser lines attached (405 nm, 491 nm, 561

nm, and 640 nm) and the 640 nm seems to be suffering from a large amount of

scattering in the light path. All of this is using the 150 x 1.45 NA lens.

Members of Olympus OSIS have flown over to look at the system tomorrow (OSIS in

Germany is where the system was developed) so hopefully they can resolve some of

the issues. they are also struggling to supply any laser safety interlocked

system which is compatible with our stage and incubation system. I think that is

going to be more difficult to rectify.



What was wrong with your system? Did you buy an alternative system? If so would

you mind letting me know which you went for. We are a bit restricted in our

choice as we really need to use the high magnification high NA lens available

from Olympus.



Colin





Dr Colin Rickman

Life Science Interface

Department of Chemistry (WP 2.03)

School of Engineering and Physical Sciences

Heriot-Watt University

Edinburgh

EH14 4AS



Tel: [phone] (Office)



http://www.lifescienceinterface.hw.ac.uk

http://www.eps.hw.ac.uk/departments/chemistry/cr.htm



On 11 May 2011, at 02:17, Jim Keen wrote:



> *****

> To join, leave or search the confocal microscopy listserv, go to:

> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

> *****

>

> Colin,

>

> Regrettably, we have had a very similar experience with this exact system.  We

recently returned it to the manufacturer after it could not be made to perform

to specs after almost 12 months.  

>

> Feel free to contact me offline for the sordid details.

>

> Jim Keen

>

> James H. Keen, Ph.D.

> Professor, Department of Biochemistry and Molecular Biology

> Jefferson Medical College

> Faculty Supervisor, Bioimaging Facility, Kimmel Cancer Center

> Thomas Jefferson University

> 233 S. 10th St., BLSB/915

> Philadelphia, PA 19107

>

> 215-503-4624 (Voice)

> 215-503-0622 (Fax)

> [hidden email]

> On May 10, 2011, at 3:20 PM, Colin Rickman wrote:

>

>> *****

>> To join, leave or search the confocal microscopy listserv, go to:

>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

>> *****

>>

>> Hi

>>

>> We recently purchased an Olympus Cell Excellence TIRF system with four

independent laser lines and the additional hardware FRAP scanner. We are having

numerous ongoing problems with both the hardware, software and support from the

UK arm of Olympus microscopes. I was wondering if anybody else has taken

delivery of such a system (with the new 4 line motorised TIRF combiner) and if

they are having problems.

>>

>> Colin

>>

>> Dr Colin Rickman

>> Life Science Interface

>> Department of Chemistry (WP 2.03)

>> School of Engineering and Physical Sciences

>> Heriot-Watt University

>> Edinburgh

>> EH14 4AS

>>

>> Tel: [phone] (Office)

>>

>> http://www.lifescienceinterface.hw.ac.uk

>> http://www.eps.hw.ac.uk/departments/chemistry/cr.htm

>>