Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
The trick to use in Photoshop to operate in RGB space where most of us
think better is to type control-y to see the result in CMYK while
operating in RGB.
Practically, to do CFP/YFP merge in RGB space would be to put the CFP in
the blue channel, the YFP in the red channel, and the maximum pixel
combination of the two channels in the green channel.
___________________________________
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On Wed, 26 Feb 2003, Glen MacDonald wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Starting fresh from the original grayscales is a fairly painless means
> of generating CMYK figures. The problems usually arise from generating
> an RGB figure then converting it to CMYK. The color conversion can be
> dreadful and affected by your color management module (if any), OS or
> application version and how far you've manipulated colors in the RGB
> colorspace. The one manner in which I've found this to work in
> Photoshop 7 is to select a CMYK printer profile for the working space,
> and 'Preserve embedded profiles' for color management policy. At least
> this works in Photoshop 7, with the Tektronix Phaser SDX Colorsync
> profile and using the Colorsync conversion engine in Apple's OSX.
> Adobe changes the location and names of these settings between software
> versions. Havent' yet compared other CMYK printer profiles or other
> color conversion engines.
>
> Regards,
> Glen
> On Wednesday, February 26, 2003, at 03:15 PM, Jacqui Ross wrote:
>
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > Hi Hugo,
> >
> > I'm not sure how you were working with the files. If you have already
> > tried
> > using the channels menu with ghastly results, read no further. If
> > not, it's
> > worth giving it a go, although it's a bit hard when I don't know what
> > your
> > images look like.
> >
> > Make a new file with the same resolution as the images with a black
> > background
> > and in CMYK colour mode. Make your original images grayscale and
> > adjust if
> > needed using the levels menu. Then go to the new black image and go to
> > the
> > channels menu. For the cyan (CFP) image, turn off all the channels
> > except cyan
> > and make sure this channel is highlighted. Go back to your cyan (CFP)
> > image,
> > select all and copy. Then go to the black image and paste the image
> > into the
> > cyan channel.
> >
> > For the YFP one, do the same thing except this time, turn all the
> > channels off
> > except yellow and paste the YFP image into the yellow channel. Then
> > turn all
> > channels on again and you will have your composite in CMYK.
> >
> > Cheers,
> >
> > Jacqui.
> >
> > "Caldas, Hugo" wrote:
> >
> >> Search the CONFOCAL archive at
> >> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >>
> >> I have a question that mind sound a little stupid and not really what
> >> this
> >> mailing list is about but....
> >>
> >> I have acquired CFP and YFP images, and am trying to use Photoshop to
> >> overlay the 2. Does anyone know how I can do this keeping the colors
> >> cyan
> >> and yellow? I have attempted CMYK but didn't take me very far. My
> >> composite
> >> images just end up looking horrible and I cannot make my point this
> >> way.
> >> Any help would be greatly appreciated.
> >>
> >> Hugo Caldas
> >> Columbus Children's Hospital
> >> Columbus, OH
> >>
> >> -----Original Message-----
> >> From: Guy Cox [mailto:[log in to unmask]]
> >> Sent: Wednesday, February 26, 2003 3:44 AM
> >> To: [log in to unmask]
> >> Subject: YFP/CFP excitation
> >>
> >> Search the CONFOCAL archive at
> >> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >>
> >> This reference recently posted to the MPLSM group is absolutely
> >> essential reading for those engaged in the current debate about
> >> MP excitation of GFPs.
> >>
> >> Essentially (and as expected since its a non-symmetrical chromophore)
> >> the excitation spectrum for the various GFP versions is a close
> >> match to the single photon spectrum (at twice the wavelength).
> >> Excitation of eGFP at 800nm is only about 20% of the excitation
> >> at the peak, 860nm. Unfortunately they don't give wtGFP but
> >> since that has a major excitation peak at 395 is could be good
> >> at 790-800 in MP for those who have non-tunable lasers.
> >>
> >> Thanks to Ben Youn for posting this.
> >>
> >> Guy
> >>
> >>> Here's a reference for TPE spectra for a couple of fluorophores,
> >>> including eGFP, eCFP, eYFP and DsRed
> >>>
> >>> Two-photon excitation action cross-sections of the autofluorescent
> >>> proteins
> >>> Blab GA, Lommerse PHM, Cognet L, Harms GS, Schmidt T
> >>> CHEMICAL PHYSICS LETTERS 350 (1-2): 71-77 DEC 14 2001
> >>>
> >>> Regards,
> >>> Ben Youn
> >>
> >> Assoc. Prof. Guy Cox, ooOOOOOOoo
> >> E.M. Unit, F09 # oOOOO | | OOOOo #
> >> University of Sydney ### OOO| | | | | |OOO ###
> >> NSW 2006, Australia ### OOO | | | | | | OOO ###
> >> Ph: 02 9351 3176 ### OO | | | | | | | | OO ###
> >> Fax: 02 9351 7682 ##### | | | | | | | | #####
> >> ==#####============================#####==
> >> http://get.to/gcc ##### #####
> >> http://guycox.cjb.net ~~#####~~~~~~~~~~~~~~~~~~~~~~~~~~~~#####~~
> >
> > Jacqueline Ross
> > Biomedical Imaging Research Unit
> > Division of Anatomy with Radiology
> > Faculty of Medical & Health Sciences
> > The University of Auckland
> > Private Bag 92019
> > Auckland, NEW ZEALAND
> >
> > Tel: 64 9 373 7599 Ext 87438
> > Fax: 64 9 373 7484
> >
> > http://www.health.auckland.ac.nz/biru/
> >
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923 USA
> (206) 616-4156
> [log in to unmask]
>
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