
Fluorescence & Flow Cytometry Tutorial
#2
Posted 27 January 2009 - 05:07 PM
#3
Posted 31 January 2009 - 09:25 AM
Fluorescence & Flow Cytometry Tutorial
http://www.invitroge.../Tutorials.html
Thank you for the link. Its very informative
Bests
pooja
~Judah Folkman
#4
Posted 07 March 2010 - 11:46 PM
#5
Posted 07 March 2010 - 11:47 PM
Fluorescence & Flow Cytometry Tutorial
http://www.invitroge.../Tutorials.html
great details, thanks...
#6
Posted 22 March 2010 - 05:29 AM
I don't think converting the mean fluorescence as measured by the flow cytometer to RFU is possible, or rather, would make sense. As the name indicates, realtive fluorescence units are no really actual, universally defined units of measurement, but merely indicate the relative intensity of the measured fluorescence as compared to each other. The mean fluorescence measured by the flow cytometer, on the other hand, is the mean of the integrated areas of all light pulses measured by the detector, and is possibly instrument specific.if any body there can tell about how to calculate relative fluorescence units out of his to gram analysis in FACS. I want to know about mean, geometric mean and CV and their conversion into more understandable format of RFU and SD?
TL;DR I think that RFU and mean fluorescence are instrument- and experiment-specific values which are meaningless without controls to which they can be compared, and can therefore not be converted. Please correct me if I'm wrong.
#7
Posted 02 February 2011 - 05:34 AM
First, you need calibration beads with known number of molecules of the fluorochrome of interest binded.if any body there can tell about how to calculate relative fluorescence units out of his to gram analysis in FACS.
#8
Posted 07 June 2012 - 03:37 AM
So is it necessary, then why?
#9
Posted 07 June 2012 - 03:46 AM
#10
Posted 10 May 2017 - 05:04 PM
Hi
I am trying to analyse pre-existing Flow Data for receptor expression, but a little confused about the controls. I believe they were all from tubes with equal amounts of cells/mL stained and a couple tubes unstained.
The controls are the confusing part...
1.) I have been given a set of Flow Cytometry data which has:
IC-PE (isotype control-pe), IC-FITC (isotype control FITC), ICAM-1and the unstained tubes.
From my understanding the unstained tubes work as my background/negative
I also been advised that my IC-FITC is also the background/negative
2.)But here is the confusing part, what is the IC-PE?
Is there a difference between IC-PE and IC-FITC? Are they both the background, treated as the negative?
3.)What does the ICAM-1 represent, would this be my positive control?
I also have other stained tubes which express positive for the receptors I am trying to find, so I do have a positive, but wondered why I have been given ICAM-1 as well?
4.)Also regards to MFI, which is best to calculate for MFI?
I have been using the median values of the IC-FITC, unstained and the tubes with the receptor staining to compare differences using an unpaired t-test, but have been advised this is incorrect.
Any clarification would be greatly appreciated please. Thank you.
#11
Posted 31 August 2018 - 02:57 PM
I found this Flow protocol that might be useful as well.
http://www.genetex.c...fluorescent.pdf