This response came from someone using the confocal microscopt in our facility (Centers for Disease Control and Prevention, Atlanta, GA): My response would be that there are a wide variety of UV-excitable probe and dyes that can be very useful in three color confocal applications. Our system has UV laser (351nm), Argon(488 nm)-Krypton(568 nm), and Neon(633 nm) lasers which give us the widest fluor capabilities available for biological applications. We do not use our UV laser as often as the Ar-Kr laser since it requires a large cooling unit which gives quite a bit of vibration related noise in images. This is a problem when one is looking at fine detail in single cells which is often the case. The other problem is the availability of good UV-fluorophores which are bright and stable. We use the Cascade, Marina and Pacific blue dyes from Molecular Probes which work reasonably well when conjugated to secondary antibodies for most applications. They are not real bright when compared to Red or Green fluors that we use. There is sometimes a problem with bleed through to the green channels if you are using a green fluor that uses a emission filter which can allow the UV fluor to be seen as green. This can also be a problem with UV counterstains like Dapi or the Hoechst dyes. I have seen the New Zeiss LSM-510 with a UV laser on it and it seems to be a little better at containing the vibration than our LSM-410 with a UV laser so that may not be a big problem depending on how your room is laid out. The advantage to having the UV in my opinion outweighs the problems if you are ever faced with the need for that third color. Of course the cost is a factor and if you don't think that you will get the use out of it that you should, then I would leave it out. You could always add it later or add one of these "personal confocal" instruments which do three color applications using a conventional visible light system for a fraction of the cost. These systems are pretty new and don't seem to have the power of a laser-based system, but you can get into one for about $50K and if it doesn't work well enough still go for the laser system later on as well. I hope this has been some help. If you are interested in more advice or have any questions, drop me a line or give me a call. Tom Voss, Ph.D. REVB Measles Confocal Group (404)639-3308 (lab)/3619 (office) > -----Original Message----- > From: GS - Jeanene Swiggett [SMTP:jswigget@carolinas.org] > Sent: Tuesday, January 20, 1998 3:43 AM > To: Cytometry Mailing List > Subject: Confocal Query > > > I am posting this query for a co-worker putting together a purchase > order > for a Confocal Microscope System. Our institution; Cannon Research > Center > at Carolinas Medical Center in Charlotte, NC; is a multiuse medical > research > center and she is interested in what various applications on a > confocal > microscope require UV excitation. I'm sure she would also appreciate > input > as to the best multi-application systems with only bright light > excitation > and those with bright light and UV excitation. Thank you for any and > all > input, Jeanene > ------------------------------------- > Name: Jeanene Swiggett > E-mail: jswigget@carolinas.org > Date: 1/20/98 > Time: 8:42:53 AM > > This message was sent by Chameleon > -------------------------------------
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