tag:blogger.com,1999:blog-8037707352330428788.post5695795527626751793..comments2024-01-26T04:59:37.784-06:00Comments on UCFlow - Flow Cytometry news, reviews, and tips.: The most sensitive Cytometer available?UC Flowhttp://www.blogger.com/profile/03187449850452376466noreply@blogger.comBlogger4125tag:blogger.com,1999:blog-8037707352330428788.post-26117880682735381632012-11-13T09:36:25.726-06:002012-11-13T09:36:25.726-06:00Thanks for your thoughts David. In principle, I a...Thanks for your thoughts David. In principle, I agree. If I could make only one comment it would be this: End-users need to test their panel of fluorochromes empirically to determine if they can in fact achieve better resolution on the ImageStream (or FlowSight). Resolving from blank using analogous dyes may not directly correlate with resolving stained cells from autofluorescence. This empowerment and encouragement to end-users of these technologies is really the point of my pontificating. Anonymoushttps://www.blogger.com/profile/07384241547190280663noreply@blogger.comtag:blogger.com,1999:blog-8037707352330428788.post-5438559504505070982012-11-12T23:00:36.406-06:002012-11-12T23:00:36.406-06:00Hi Ryan,
Thanks for the writeup on this, sorry I&...Hi Ryan,<br /><br />Thanks for the writeup on this, sorry I'm so late to the party. I understand your dissatisfaction with 8 peak Rainbow beads as sensitivity standards but they do have several useful characteristics:<br /><br />1. They have a true unlabeled population and a closely-spaced dim population. It can be surprisingly difficult to find a truly "dark" bead. A lot of unlabelled beads that we've looked at are actually dimly fluorescent for one reason or another.<br /><br />2. The brightest population in the set can be adjusted to lie just below the saturation point of the detector, allowing measurement of sensitivity at the low end using settings that provide maximum dynamic range. Any system can look good at the low end by cranking up detector voltage and/or laser power but by constraining the high end to lie below saturation, you get an apples-to-apples means of measuring sensitivity across instruments under the most demanding conditions. <br /><br />3. The broadband ("white") emission of these beads makes them very convenient to use for measuring deep red sensitivity with 488 excitation without having to resort to things like home-brew beads capturing PE-Cy7 labeled antibodies, which can be difficult to reproduce. <br /><br />As you point out, we're not shy about the claims we make for the sensitivity of our instruments, but always with data to back them up. I'd be very interested to see more results from your independent cross-platform testing.<br /><br />Best,<br />DavidDavid Basijihttp://www.amnis.comnoreply@blogger.comtag:blogger.com,1999:blog-8037707352330428788.post-59863681489384552532011-11-29T12:12:53.643-06:002011-11-29T12:12:53.643-06:00I certainly agree with your comments, and I believ...I certainly agree with your comments, and I believe Amnis has been pretty clear that the ImageStream is not a conventional flow cytometer. I guess I'm mostly looking at marketing and perception of the Flowsight as an alternative to a flow cytometer of similar capability and price-point. The claims that were being made and the data used to support such claims (i.e. 8-peak bead resolution) are not really valid. If I were marketing the instrument, I'd focus most of my attention on the world of information obtained with the addition of imagery and how that can allow a researcher to make more definitive conclusions for some applications. Thanks for the Comment!Anonymoushttps://www.blogger.com/profile/07384241547190280663noreply@blogger.comtag:blogger.com,1999:blog-8037707352330428788.post-58152489449395167492011-11-29T10:12:06.067-06:002011-11-29T10:12:06.067-06:00The Imagestream certainly has capabilities that th...The Imagestream certainly has capabilities that that of conventional cytometer does not. Therefore I support and always have that you should never always depend on data from one type of application/instrument and should always complement it with that of another to develop a well rounded publication and truly understand what is occuring in your experiment. Good research in not "1-dimensional".Anonymousnoreply@blogger.com