Hi Tim,
The spectral website: http://www.spectra.arizona.edu/ will show you specifically what the "efficiency" of absorbance is for any dye you might be considering. Simply place the cursor over the curve at any wavelength to find out the relative absorbance. This can be used to roughly pre-test dyes vs. excitation sources so you have a rational basis for considering which laser might do what you need.
Good luck.
C
Carl A. Boswell
520-954-7053
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Tim Feinstein
Sent: Thursday, November 10, 2011 6:09 AM
To: [log in to unmask]
Subject: Re: FRET in the time of DPSS
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
Hi guys (and Guy),
Many thanks for the feedback. I like the 505 laser idea in particular. As long as it excites 488 fluorophores well enough it could be exactly what I need. Does anybody have experience with using that in place of a 488 line?
All the best,
Tim
Sent from my iPad
On Nov 10, 2011, at 1:57 AM, Guy Cox <[log in to unmask]> wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The Sapphire series lasers are OPS (Optically Pumped Semiconductor)
> not DPSS but, you are right, they do indeed have a 514nm version. I
> don't know why I didn't find it, but did find the 505, when I searched
> this morning. I still think that the 505 might be exactly what
> Timothy needs, though, since he doesn't want to buy an extra laser.
>
> I have also been contacted off-list and told that Spectra-Physics have
> a 515nm DPSS which is close enough to make no difference. I've no
> idea what the lasing crystal is.
>
> 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) Australian Centre for
> Microscopy & Microanalysis, Madsen Building F09, University of Sydney,
> NSW 2006
>
> Phone +61 2 9351 3176 Fax +61 2 9351 7682
> Mobile 0413 281 861
> ______________________________________________
> http://www.guycox.net
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[log in to unmask]]
> On Behalf Of samuel connell
> Sent: Thursday, 10 November 2011 1:57 PM
> To: [log in to unmask]
> Subject: Re: FRET in the time of DPSS
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Industry Response:
>
> There absolutely is a 514 DPSS available from 20 mW up to 150 mW
> available in the Sapphire form factor from Coherent. We use this
> laser, often in conjunction with the 445 CUBE in our LaserStack for
> CFP/YFP (and their newer brothers and sisters in FP evolution) imaging
> for FRET or otherwise.
>
> ------------------------
> Samuel A. Connell
> Sales Manager
> Pacific Region-North America
> Intelligent Imaging Innovations, Inc
> 3250 Ocean Park Blvd, Suite 202
> Santa Monica, CA 90405
> Cell: (858) 692-4510
> [log in to unmask]
>
>
> On Wed, Nov 9, 2011 at 4:21 PM, Damir Sudar <[log in to unmask]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>>
> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u
> mn .edu/cgi-bin/wa?A0=confocalmicroscopy>
>> *****
>>
>> See: http://www.cobolt.se/**coboltfandango515nm.html?**
>>
> gclid=CPCOueXlqqwCFQyEhwodQn-**aAw<http://www.cobolt.se/coboltfandango
> 51 5nm.html?gclid=CPCOueXlqqwCFQyEhwodQn-aAw>
>>
>> No connection, just considering about switching away from our old Ar
> Ion
>> as well.
>>
>> - Damir
>>
>>
>> On 11/9/2011 3:36 PM, Guy Cox wrote:
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>>
> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u
> mn .edu/cgi-bin/wa?A0=confocalmicroscopy>
>>> *****
>>>
>>> Are you sure there is a 514nm DPSS? I think that wavelength would
> have
>>> to be an OPS and I don't know anyone who makes one. Coherent do
>>> make
> a
>>> 505nm OPS which is intended to stand in for both the 488& 514 lines
> of
>>>
>>> an Argon laser, and I rather suspect that this could be exactly what
> you
>>> need. But I have no experience of it. (I do have a Coherent 488nm
>>> OPS).
>>>
>>> Guy
>>>
>>> Optical Imaging Techniques in Cell Biology
>>> by Guy Cox CRC Press / Taylor& Francis
>>>
>>>
> http://www.guycox.com/optical.**htm<http://www.guycox.com/optical.htm>
>>> ______________________________**________________
>>> Associate Professor Guy Cox, MA, DPhil(Oxon) Australian Centre for
>>> Microscopy& Microanalysis,
>>>
>>> Madsen Building F09, University of Sydney, NSW 2006
>>>
>>> Phone +61 2 9351 3176 Fax +61 2 9351 7682
>>> Mobile 0413 281 861
>>> ______________________________**________________
>>> http://www.guycox.net
>>>
>>>
>>>
>>> -----Original Message-----
>>> From: Confocal Microscopy List
> [mailto:CONFOCALMICROSCOPY@**LISTS.UMN.EDU<[log in to unmask]
> EDU>
>>> ]
>>> On Behalf Of Tim Feinstein
>>> Sent: Thursday, 10 November 2011 7:34 AM
>>> To: [log in to unmask]**EDU
> <[log in to unmask]>
>>> Subject: FRET in the time of DPSS
>>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>>
> http://lists.umn.edu/cgi-bin/**wa?A0=confocalmicroscopy<http://lists.u
> mn .edu/cgi-bin/wa?A0=confocalmicroscopy>
>>> *****
>>>
>>> Hello all,
>>>
>>> We want to spec a four-laser launch for a new live cell system that
> will
>>> handle both CFP/YFP FRET and red/green imaging. However, I am sad
> to
>>> see that gas lasers are no longer speccable and so the freebie 514
> laser
>>> line is gone. We would therefore have to spec a 514 DPSS and forego
> the
>>> far-red line.
>>>
>>> I was wondering whether there is a way to do more (or at least the
> same)
>>> with less. 488 nm excites YFP well enough, so in theory I could
> image
>>> CFP/YFP using a scan head dichroic with cutouts for 442 and 488 nm
> laser
>>> lines. In my experience 442 nm laser excitation (via TIRF) causes
>>> negligible YFP excitation and 488 nm does not excite CFP, so it is
>>> possible that I could gain speed by passing everything through a
> single
>>> broad bandpass filter (e.g., 455-550 nm) and alternate excitations.
>>> Assuming that cross-talk is not a problem, the most significant cost
>>> would be that I lose a decent chunk of CFP emission to the scan head
>>> dichroic, but in return I gain a 641 nm laser.
>>>
>>> Has anyone tried this? Any feedback on or off-list would be much
>>> appreciated.
>>>
>>> Thanks and all the best,
>>>
>>>
>>> TF
>>>
>>> Timothy Feinstein, PhD
>>> Postdoctoral Fellow
>>> Laboratory for GPCR Biology
>>> Dept. of Pharmacology& Chemical Biology
>>>
>>> University of Pittsburgh, School of Medicine BST W1301, 200 Lothrop
>>> St.
>>> Pittsburgh, PA 15261
>>>
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>>
>> --
>> Damir Sudar - Staff Scientist and Deputy for Technology Lawrence
>> Berkeley Laboratory / Life Sciences Division One Cyclotron Road, MS
>> 977, Berkeley, CA 94720, USA
>> T: 510/486-5346 - F: 510/486-5586 - E: [log in to unmask]
>> WWW: http://www.lbl.gov/lsd/People_**&_Organization/Scientific_**
>>
> Staff_Directory/Sudar_Lab.html<http://www.lbl.gov/lsd/People_&_Organiz
> at ion/Scientific_Staff_Directory/Sudar_Lab.html>
>>
>
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