***************"Commercial Response" ******************
Dear Inna,
Ideally it is desired to provide very high degree of blocking (several
orders of magnitude of blocking) of the unwanted laser lines to avoid
interference patterns in the emission channel. Both the excitation
filter and the emission filter contribute to this blocking of unwanted
laser lines. For extremely demanding applications such as imaging of
single molecules, it may be even desirable to provide additional
blocking by using a suitable notch or a long pass filter in the emission
channel
(http://www.semrock.com/Data/Documents/PerfectingTIRF_0109_Reprint.pdf).
This is generally not true for a dichroic mirror, as they are not
specifically designed for blocking.
For your application, it may be worthwhile considering from following
Semrock filter for imaging CFP & YFP.
1) CFP-2423A set
(http://www.semrock.com/Catalog/SetDetails.aspx?SetBasePartID=21&Categor
yID=16). Note that the emission filter of this filter set,
FF01-483/32-25 is designed to provide >OD 6 blocking at 458 nm.
2) YFP-2427A set
(http://www.semrock.com/Catalog/SetDetails.aspx?SetBasePartID=25&Categor
yID=16). Note that the emission filter of this filter set,
FF01-542/27-25 is designed to provide >OD 6 blocking at 514 nm.
3) Alternatively, a steep edge long pass filter in the emission channel,
such as Semrock's LP02-458RS-25
(http://www.semrock.com/Catalog/Detail.aspx?FilterPartID=150&CategoryID=
69), which is designed to provide > OD 6 blocking at 458 nm when used in
conjunction with your existing filters should be able to eliminate
interference patterns in this channel.
Please contact me offline at [log in to unmask], if you have
questions.
Regards,
Prashant
Prashant Prabhat, Ph.D.
Applications Specialist
Semrock
A Unit of IDEX Corporation
3625 Buffalo Road, Suite 6
Rochester NY 14624
Email: [log in to unmask]
Phone: 585-594-7064
Toll Free: 866-SEMROCK
Fax: 585-594-7095
http://www.semrock.com <http://www.semrock.com/>
The Standard in Optical Filters for Biotech & Analytical Instrumentation
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extensive inventory now!
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________________________________
From: Confocal Microscopy List [mailto:[log in to unmask]]
On Behalf Of Inna Slutsky
Sent: Tuesday, May 26, 2009 1:57 AM
To: [log in to unmask]
Subject: CFP detection under 458 excitation
Recently we upgraded our Olympus FV-300 confocal system and purchased
filters for CFP/YFP detection for FRET applications.
The problem is that the 465-495 nm band pass filter (Chroma) showed
direct excitation of the 458 line (multi-line argon laser) and a heavy
interference with direct reflection from the glass. Chroma claims that
it only passes 0.3% of 458nm, and theoretically it is supposed to be OK
for 458 nm excitation.
I wonder if someone has encountered a similar problem and may offer a
better solution for detection of CFP signal (together with YFP under 514
nm excitation).
Thanks,
Inna.
--------------------------------------------------------
Inna Slutsky, PhD
Dept. of Physiology and Pharmacology
Sackler Faculty of Medicine, room 511
Tel Aviv University
Ramat Aviv, Tel Aviv 69978, Israel
phone: +972 3 640 6021
e-mail: [log in to unmask]
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