Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Christophe, I would approach this from an empirical angle. Look at both sets! Your Zeiss rep should be able to acquire them so that you can demo both Zeiss and Semrock filters. Semrock and Chroma will provide spectral characteristics of their products if you ask. Moreover, if you find their stock filter sets unsatisfactory, both Semrock and Chroma are able to custom make the filters to your specs.
Brian D Armstrong PhD
Light Microscopy Core Manager
Beckman Research Institute
City of Hope
1450 E Duarte Rd
Duarte, CA 91010
626-359-8111 x62872
http://www.cityofhope.org/SharedResources/LightMicroscopy
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Christophe Leterrier
Sent: Tuesday, February 13, 2007 5:32 AM
To: [log in to unmask]
Subject: Filters for eGFP/mCherry imaging
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi all,
Not a confocal-related question, but there are very good specialists of
widefield imaging here !
I'm trying to optimize a fast 3D widefiled/TIRF setup for two-color
imaging of eGFP/mCherry.
I got 488nm and 561nm laser diodes for TIRF.
I have a DG4 wavelength switcher for epifluorescence excitation. I don't
have an emission filterwheel, so I need a "Pinkel" filter set for rapid
imaging of eGFP and mCherry. It consists of two single band excitation
filters (in the DG-4) and a cube with a dual band dichroïc and a dual
band emission filter.
I want to be as much "light efficient" as possible, so I looked at the
different possibilities offered with "high transmittance" filter sets
from Zeiss (the scope is a Zeiss), Chroma and Semrock.
You can see the characteristics of the different filters I'm considering
on this page :
http://spreadsheets.google.com/pub?key=p1_lbCUheIRM4pjbGruaLKw
The problem is, from Zeiss and Semrock I have to choose between
GFP+DsRed (a bit on the lower wavelength side for mCherry) and GFP+HcRed
(a bit on the higher wavelength side). Which one is the best compromise ?
There is also the Chroma double band whih seems perfect, but I don't
know if it's a Pinkel set and I also don't know the dichroïc
characteristics (they're not on their website).
In case you're wondering, I've made a spectrum graph of both proteins
available here (data from R. Tsien website) :
http://docs.google.com/Doc?id=dfsfzj8m_0dttt6h
Let me know what you think !
"EMF <COH.org>" made the following annotations.
------------------------------------------------------------------------------
SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender.
==============================================================================
|
