changing filters on Leica TCS SP2 system

[Leigh Silvester]

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Hi
Has anyone got any experience of replacing the filter cubes for a TCS
SP2 system?

Am quite impressed with what I have heard about Semrock filters and am
thinking of getting one
for Texas Red as I am not convinced that the triple dichroic I have is
letting through enough light.

Regards

Leigh Silvester
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University of Nottingham
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[vb-2]

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I doubt that Semrock's filters/dichroics have AR-coating for the laser
applications (very few - for the very very broad market)

vitaly

----- Original Message -----
From: "Silvester Leigh" <[hidden email]>
To: <[hidden email]>
Sent: Monday, October 15, 2007 10:34 AM
Subject: changing filters on Leica TCS SP2 system

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi
> Has anyone got any experience of replacing the filter cubes for a TCS
> SP2 system?
>
> Am quite impressed with what I have heard about Semrock filters and am
> thinking of getting one
> for Texas Red as I am not convinced that the triple dichroic I have is
> letting through enough light.
>
> Regards
>
> Leigh Silvester
> ==================================================================
> Division of Animal Physiology
> University of Nottingham
> Sutton Bonington Campus
> Loughborough
> Leicestershire
> LE12 5RD
> T: [phone] x18736
> F: [phone]
> E: [hidden email]
> ==================================================================
> Do you need to print this email?
> ==================================================================
>
>
> This message has been checked for viruses but the contents of an
> attachment
> may still contain software viruses, which could damage your computer
> system:
> you are advised to perform your own checks. Email communications with the
> University of Nottingham may be monitored as permitted by UK legislation.
>

[Steffen Dietzel]

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Just for clarification, Which filters do you mean? If it is cubes,
that would be the non-confocal part? Or the confocal beamsplitter
where some systems have the AOBS?

Steffen

At 17:34 15.10.2007, you wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi
>Has anyone got any experience of replacing the filter cubes for a TCS
>SP2 system?
>
>Am quite impressed with what I have heard about Semrock filters and am
>thinking of getting one
>for Texas Red as I am not convinced that the triple dichroic I have is
>letting through enough light.
>
>Regards
>
>Leigh Silvester
>==================================================================
>Division of Animal Physiology
>University of Nottingham
>Sutton Bonington Campus
>Loughborough
>Leicestershire
>LE12 5RD
>T: [phone] x18736
>F: [phone]
>E: [hidden email]
>==================================================================
>Do you need to print this email?
>==================================================================
>
>
>This message has been checked for viruses but the contents of an attachment
>may still contain software viruses, which could damage your computer system:
>you are advised to perform your own checks. Email communications with the
>University of Nottingham may be monitored as permitted by UK legislation.

[Leigh Silvester]

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Steffen

I mean the confocal beamsplitter where some systems have the AOBS.
Possibly what I have in mind is not really practicable. and perhaps as
someone else pointed out
the semrock filters may not be suitable for confocal. There is also the
software side of the equation
as well...

Just investigating options.

Regards

Leigh Silvester
==================================================================
Division of Animal Physiology
University of Nottingham
Sutton Bonington Campus
Loughborough
Leicestershire
LE12 5RD
T: [phone] x18736
F: [phone]
E: [hidden email]
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Do you need to print this email?
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[Michael Weber-4]

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Steffen,

as far as I heard from Leica, changing the dichroics to hard coated ones
is not possible - they are too thick for the standard holder. It
probably also introduces a shift due to the thicker material, so the
emission beam will go somewhere else. You would need to do it on your
own risk, I guess.

cheers,
Michael


Silvester Leigh wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Steffen
>
> I mean the confocal beamsplitter where some systems have the AOBS.
> Possibly what I have in mind is not really practicable. and perhaps as
> someone else pointed out
> the semrock filters may not be suitable for confocal. There is also the
> software side of the equation
> as well...
>
> Just investigating options.
>
> Regards
>
> Leigh Silvester
> ==================================================================
> Division of Animal Physiology
> University of Nottingham
> Sutton Bonington Campus
> Loughborough
> Leicestershire
> LE12 5RD
> T: [phone] x18736
> F: [phone]
> E: [hidden email]
> ==================================================================
> Do you need to print this email?
> ==================================================================
>
>
> This message has been checked for viruses but the contents of an attachment
> may still contain software viruses, which could damage your computer system:
> you are advised to perform your own checks. Email communications with the
> University of Nottingham may be monitored as permitted by UK legislation.

[Carol Norris]

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Hi Leigh,

The beam splitters used by the SP2 don't live in cubes, rather they are mounted in a frame in the scanner. I don't know if they are standard sizes or not, but I personally would not want to disassemble anything in the scanner! In my experience, the challenge with probes like Texas Red is the 543 laser,  which is really low power (1.2 mW compared to 20 mW for the 488 line) in addition to being off-peak for Texas Red excitation. If you are not using the 633 laser line for another fluor, try using the DD488/543 instead of the triple dichroic - the TD will block some of the Texas Red emission. I have copies of the transmission curves for the beam splitters and can send them along if you'd like - I've found them useful for figuring things like this out. 

Regards,

Carol Norris

On Oct 15, 2007, at 11:34 AM, Silvester Leigh wrote:

Search the CONFOCAL archive at

http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi

Has anyone got any experience of replacing the filter cubes for a TCS

SP2 system?

Am quite impressed with what I have heard about Semrock filters and am

thinking of getting one

for Texas Red as I am not convinced that the triple dichroic I have is

letting through enough light.

Regards

Leigh Silvester

==================================================================

Division of Animal Physiology

University of Nottingham

Sutton Bonington Campus

Loughborough

Leicestershire

LE12 5RD

T: [phone] x18736

F: [phone]

E: [hidden email]

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Do you need to print this email?

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This message has been checked for viruses but the contents of an attachment

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Carol E. Norris, Ph.D

Facility Scientist

Flow Cytometry/Confocal Microscopy Facility

Biotechnology/Bioservices Center

University of Connecticut Unit 3149

91 N. Eagleville Rd

Storrs, CT 06269-3149

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