Re: How to define the presence/absence of an object in immunofluorescence

Posted by Jens on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Two-photon-microscope-questions-tp7583010p7583028.html

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Yes, however I do not have a reference of the top of my head.

Jens

Jens-B. Bosse
+1-609-216-6388

> On Nov 30, 2014, at 13:38, AroPro <[hidden email]> wrote:
>
> Thanks a lot, Jens. You mean the intensity should be >= (mean background + 2*SD of the background)? So the difference will be statistically significant?
> Is there any reference I can refer to?
> Best,
> Aro
>
>> Date: Sun, 30 Nov 2014 13:24:48 -0500
>> From: [hidden email]
>> Subject: Re: How to define the presence/absence of an object in immunofluorescence
>> To: [hidden email]
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Hi,
>>
>> 2x the SD over mean background is a good rule of thumb.
>>
>> Jens
>>
>> Jens-B. Bosse
>> +1-609-216-6388
>>
>>> On Nov 30, 2014, at 12:55, PengKe <[hidden email]> wrote:
>>>
>>> Dear lister:
>>> I'm recently puzzled by a question that I felt might have puzzled some of you as well and I would like to receive some suggestions and opinions from you.
>>> Question: In immunofluorescence analysis, sometimes we detected objects that showed very weak signals, so how can we decide whether the signal is really there or not?
>>> To make the question a little more specific, I will give some artificial numbers. Let's say the background has the signal intensity of 2000 and the potential object showed a signal intensity of 2200. The saturating intensity of the camera is 65535. In this case can one claim the 2200 signal represent a real object?
>>> A related question might be: is there a golden standard about how much higher a signal intensity needs to be above the background to be defined as an object?
>>> I'm looking forward and I'll be very grateful to your opinions.
>>> Best wishes,
>>> Aro                        
>