Re: background removal in fluorescence

Posted by Tony Collins-2 on Sep 20, 2005; 1:53pm
URL: http://imagej.273.s1.nabble.com/background-removal-in-fluorescence-tp3704817p3704824.html

I agree with Pedro here. My only other comment is with some older
cameras, the background is not even so subtracting a single value for
the backround is not correct and a 'black-field' subtraction may be the
better way.

If it's a timecourse you may need to take a measure of background for
each timepoint too.

Tony

Pedro J Camello wrote:

> In my opinion (my main experience is ca2+ signals in living cells) the
> best method is to acquire a background image in an area of the coverslip
> where there are no cells, using it to substract to the series of images.
> Sometimes this is difficult if the preparation is too "dense", and if
> you have to move too far from the area of interest the plane of focus
> could be different. Now I´m removing to the entire image the average
> fluorescence from an irregular ROI drawn in the "empty" area surrounding
> the cells.
>
> Martin Wessendorf wrote:
>
>> Dear ImageJ folks--
>>
>> Has anyone come up with a good method for background subtraction in
>> images obtained using fluorescence microscopy?  In my hands, the
>> methods that work for brightfield don't work for fluorescence.
>>
>> Thanks!
>>
>> Martin

--
Tony Collins, Ph.D.
Facility Manager
Wright Cell Imaging Facility
Toronto Western Research Institute
13-407 McLaughlin Pavilion
399 Bathurst Street
Toronto, ON. M5T 2S8
tel. [phone] fax: [phone]
http://www.uhnresearch.ca/wcif

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