Spectra of PerkinElmer UltraView dichroics

[Ian Dobbie]

*****
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 Everyone.

We have two Perkin Elmer Ultraview's and are trying to get spectra for
the dichroics without dismantelling them. We have very vauge
specificasions, just giving the laser passbands eg:


Dichroics
 1: 405/488/561/640
 2: 405/440/514/640
 3: 405/440/488


but no other data. I know that Semrock sell dichroics for the Yokogawa
spinnging disks but none of those seem to match the laser wavelengths
above.

Any suggestions short of taking them out and using a spectrometer?

Thanks,

Ian

[Zdenek Svindrych-2]

*****
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 Ian,

the older CSUs *may* have been fitted with these beamsplitter filters (13
mm x 15 mm size):
https://www.semrock.com/FilterDetails.aspx?id=Di01-T405/488/568/647-W1
https://www.semrock.com/FilterDetails.aspx?id=Di01-T457/514/647-13x15x0.5
(works also with 405 line, but there is no emission band between 405 nm and
450 nm)
https://www.semrock.com/FilterDetails.aspx?id=Di01-T488-13x15x0.5 (yes,
single band covering 405, 440, 488 lines, might work OK with DAPI, but not
great for BFP)

No guarantee, your dichroics may be completely different!

CSU-W1 uses bigger beamsplitters (22.2 mm x 25.0 mm). Our Andor Dragonfly
uses one of these, the passbands are listed as follows: 400-410nm,
483-493nm, 557-563nm, 633-643nm, but I have no curve.

Obviously, the spinning disk dichroics are very specific in that they are
transparent for the laser wavelengths and reflect the fluorescence; all
other regular dichroics work the other way round.

Best, zdenek

On Wed, Feb 13, 2019 at 4:41 AM Ian Dobbie <[hidden email]>
wrote:

> *****
> 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 Everyone.
>
> We have two Perkin Elmer Ultraview's and are trying to get spectra for
> the dichroics without dismantelling them. We have very vauge
> specificasions, just giving the laser passbands eg:
>
>
> Dichroics
>  1: 405/488/561/640
>  2: 405/440/514/640
>  3: 405/440/488
>
>
> but no other data. I know that Semrock sell dichroics for the Yokogawa
> spinnging disks but none of those seem to match the laser wavelengths
> above.
>
> Any suggestions short of taking them out and using a spectrometer?
>
> Thanks,
>
> Ian
>

--
--
Zdenek Svindrych, Ph.D.
Research Associate - Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth

[Ian Dobbie]

*****
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.
*****

Zdenek Svindrych <[hidden email]> writes:

> the older CSUs *may* have been fitted with these beamsplitter filters (13
> mm x 15 mm size):
> https://www.semrock.com/FilterDetails.aspx?id=Di01-T405/488/568/647-W1
> https://www.semrock.com/FilterDetails.aspx?id=Di01-T457/514/647-13x15x0.5
> (works also with 405 line, but there is no emission band between 405 nm and
> 450 nm)
> https://www.semrock.com/FilterDetails.aspx?id=Di01-T488-13x15x0.5 (yes,
> single band covering 405, 440, 488 lines, might work OK with DAPI, but not
> great for BFP)


Thanks for the hints, we have something slightly different. After
testing it is clear our system has

Semrock Di01-T405/488/568/647
Semrock Di01-T488
Semrock Di01-T442/514/647

Why these are labelled

1: 405/488/561/640
2: 405/440/514/640
3: 405/440/488

is anybodies guess.

I case anyone else is interested in this I got passable spectra by
removing the filter wheel dropping a 1/2" 45degree prism down into the
slot left by the filter wheel. Then used the halogen transmission light
with no condenser or objective and a USB spectrometer to get the
spectra.

As there is no blank position I just read the halogen spectra before the
objective and arbitrarily scaled it to get sensible-ish results. Short
and long wavelength stuff seems to fall off rather than be flat but it
is quite clear if the holes in the dichroic match up or not.

Just for completeness our filters appear to be (all from Semrock)

FF01-530/55
Em01-R405/568
Em01-R442/647
Em01-R514 - not sure about this one, ours cuts off earlier at long wavelengths
Em01-R488/568
Em01-R442/514/647

With one empty, two blocked and one a polariser in a 10 position wheel.

Thanks for everyones help with this.

Ian