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Thank you both; I appreciate the advice. Great to know I have a couple of options.
Enjoy your weekends.
Neil
From: "Periasamy, Ammasi (ap3t)" <[log in to unmask]>
Sent: Nov 19, 2015 11:21 PM
To: [log in to unmask]
Subject: Re: Autofluorescence in PFRET
Hello Neil
As Sheng mentioned you can use the spectral imaging approach. Or you can use the unlabeled cells to collect the signal to use for background subtraction. That will take care. The background subtraction should remove any autofluorescence or detector noise involved in fluorescence imaging.
Hope this helps.
Dr. Ammasi Periasamy
Professor & Center Director
http://www.kcci.virginia.edu/people/profile/ap3t
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of yuansheng.sun
Sent: Thursday, November 19, 2015 9:27 PM
To: [log in to unmask]
Subject: Re: Autofluorescence in PFRET
Then, it would be better to use spectral FRET. Measure the reference spectrum of the autofluorescence from unlabeled sample. Use linear unmixing to remove autofluorescence.
Sheng
Sent from my T-Mobile 4G LTE Device
-------- Original message --------
From: "Anthony, Neil" <[log in to unmask]>
Date:11/19/2015 3:06 PM (GMT-06:00)
To: [log in to unmask]
Cc:
Subject: Autofluorescence in PFRET
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Hi all, I hope the science treats you well.
I was wondering if anybody's had any experience with autofluorescence when using PFRET, or PeriFRET as I like to call it. I understand that autofluorescence is not accounted for in the algorithm?
Peri, I hope all is well with you. Can I ask for some pointers on how think about the system if autofluorescence is the mix?
Thanks in advance for your time.
Neil
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