UV lines on Yokogawa CSU-X1 Head

[Renato A. Mortara]

Hi !

 

Does any one has experienced using UV lines , 353 and/or 363 nm with the Yokogawa CSU-X1 Head. Yokogawa claim that in the range of 400 - 700 their microlenses work fine, but have no experience with lower wavelengths.

 

Any input (pros + cons) will be greatly appreciated.

 

All the best

 

Renato

 

Renato A. Mortara

Parasitology Division

UNIFESP - Escola Paulista de Medicina

Rua Botucatu, 862, 6th floor

São Paulo, SP

04023-062

Brazil

Phone: [phone]

Fax:     [phone]

email: [hidden email]

home page: www.ecb.epm.br/~ramortara

[Richard Berman]

Dear Renato:

Your timing is pretty good. Over the next few months we intend to
explore the potential of using the Yokogawa CSU at 375nm. For our
testing we will use the CSU-10 although the results should apply to the
X1 as well. Although the wavelength is not as low as 355nm, it is lower
than any previous work of which we are aware. You are correct to
question the transmission of the disks but there are other concerns too,
mostly on the excitation path. The transmission of the standard fiber
will be very poor at 375nm and could have a shorter lifetime when used
with such low wavelengths. We will investigate other fiber materials. We
are concerned that the chromatic shift in the collimating lenses and the
lenslets themselves could introduce problems.  In addition, there are
the usual concerns about the transmission efficiency of the scope and
objectives. A special CSU dichroic will be required to make this
possible too.

 From our side we would like to hear from you or other CSU users about
the potential benefits of 375nm excitation light. For your application
is it necessary to get down to 355nm or is 375nm useable? Operating at
355nm may be possible although it is not on our current test plans.

Regards, Richard

--
Richard Berman
[hidden email]

Spectral Applied Research
9078 Leslie St., Unit 11
Richmond Hill, Ontario
L4B 3L8

Renato Mortara wrote:

> Hi !
>  
> Does any one has experienced using UV lines , 353 and/or 363 nm with
> the Yokogawa CSU-X1 Head. Yokogawa claim that in the range of 400 -
> 700 their microlenses work fine, but have no experience with lower
> wavelengths.
>  
> Any input (pros + cons) will be greatly appreciated.
>  
> All the best
>  
> Renato
>  
> Renato A. Mortara
> Parasitology Division
> UNIFESP - Escola Paulista de Medicina
> Rua Botucatu, 862, 6th floor
> São Paulo, SP
> 04023-062
> Brazil
> Phone: [phone]
> Fax:     [phone]
> email: [hidden email] <mailto:[hidden email]>
> home page: www.ecb.epm.br/~ramortara

--
Richard Berman
Spectral Applied Research
9078 Leslie St., Unit 11
Richmond Hill, Ontario
L4B 3L8

905-326-5040 ext. 444

www.spectral.ca

[Mark Cannell]

Hi

To raise another concern, what about  UV damage to the micro lenses?

Mark.

Richard Berman wrote:

> Dear Renato:
>
> Your timing is pretty good. Over the next few months we intend to
> explore the potential of using the Yokogawa CSU at 375nm. For our
> testing we will use the CSU-10 although the results should apply to
> the X1 as well. Although the wavelength is not as low as 355nm, it is
> lower than any previous work of which we are aware. You are correct to
> question the transmission of the disks but there are other concerns
> too, mostly on the excitation path. The transmission of the standard
> fiber will be very poor at 375nm and could have a shorter lifetime
> when used with such low wavelengths. We will investigate other fiber
> materials. We are concerned that the chromatic shift in the
> collimating lenses and the lenslets themselves could introduce
> problems.  In addition, there are the usual concerns about the
> transmission efficiency of the scope and objectives. A special CSU
> dichroic will be required to make this possible too.
>
> From our side we would like to hear from you or other CSU users about
> the potential benefits of 375nm excitation light. For your application
> is it necessary to get down to 355nm or is 375nm useable? Operating at
> 355nm may be possible although it is not on our current test plans.
>
> Regards, Richard
>

[Gary Yellen]

For using UV excitation with the SD (or in any confocal scope), I would be
concerned about the chromatic aberration in the objectives.  Some of the new
objectives from many mfrs are now corrected down to 405 nm; most objectives
are not even this good.  Even without the problem of % transmission in the
UV, if the excitation wavelength is focused in a different plane than the
emission wavelength, the pinhole will reject much of the emitted light.

[Guy Cox]

I don't really know how the microlenses are made, but I assume it's by
some semiconductor-type process.  In which case, they might well be
silica, which will transmit UV OK.  Whatever they are, it would seem
unlikely that they have any chromatic correction.  I'd guess that they
would be designed around 488nm, so 365 would be out by 123nm -
equivalent to 611 in the visible direction.  Thus if the system will
work with a He-Ne (635) it should also work with 365.  

As Gary says, the objective is likely to be a problem, but you can buy
violet corrected objectives which should help quite a bit.  The other
snag is that these fancy objectives tend to be 60x, whereas I understand
the Yokogawa head is designed around 100x.  Confocality will therefore
be worse at 60x (pinholes too large) but on the flip side there will be
a bit more latitude for aberrations.

                                           Guy



Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Gary Yellen
Sent: Friday, 13 November 2009 12:37 AM
To: [hidden email]
Subject: Re: UV lines on Yokogawa CSU-X1 Head

For using UV excitation with the SD (or in any confocal scope), I would
be concerned about the chromatic aberration in the objectives.  Some of
the new objectives from many mfrs are now corrected down to 405 nm; most
objectives are not even this good.  Even without the problem of %
transmission in the UV, if the excitation wavelength is focused in a
different plane than the emission wavelength, the pinhole will reject
much of the emitted light.

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