Dear ImageJ,
I have a question how to use the ImageJ program. I have labeled cells with a fluorescence dye. I let the cells in culture about several days. Than I made pictures with the fluorescence microscope on day 1, 3 and 5 after staining. Now I want to messure and compare the fluorescence intensity signal over these days and furthermore I want to examine whether there is any weakening. So, can you say me how to to this with the ImageJ program? Thank you. With kind regards, Anette Christ _____________________________________________________________________ Der WEB.DE SmartSurfer hilft bis zu 70% Ihrer Onlinekosten zu sparen! http://smartsurfer.web.de/?mc=100071&distributionid=000000000066 |
Hi Anette,
the are a couple of ways to measure intensity. The most simple way is to load an image, point the mouse to the pixel of interest and check the signal intensity in the main tool bar. This could be a first test if the images where acquired in the range of the detector (=if nothing it overexposed). Did you acquire all images under the same condition (excitation power, filter setup, exposure time)? Otherwise it will be tricky to compare them. You can also measure regions by drawing i.e. a circle in the cell and go for "Analyze" > "Measurements". There are also plugins/macros to automate this, but a couple of manual steps before are probably go to find the best way. cheers, Michael > Dear ImageJ, > I have a question how to use the ImageJ program. I have labeled cells with > a fluorescence dye. I let the cells in culture about several days. Than I > made pictures with the fluorescence microscope on day 1, 3 and 5 after > staining. Now I want to messure and compare the fluorescence intensity > signal over these days and furthermore I want to examine whether there is > any weakening. > So, can you say me how to to this with the ImageJ program? > > Thank you. > > With kind regards, > Anette Christ |
I would emphasize Michael's point about the range of intensity. You
must make sure that your brightest image is within the 0-255 range of the 8-bit image (for example). You will not be able to compare two images each of which is so bright that it exceeds that limit. Joel Date sent: Mon, 24 Sep 2007 11:54:13 +0200 Send reply to: [hidden email] From: Michael Weber <[hidden email]> Subject: Re: fluorescence intensity To: [hidden email] > Hi Anette, > > the are a couple of ways to measure intensity. The most simple way is to > load an image, point the mouse to the pixel of interest and check the > signal intensity in the main tool bar. This could be a first test if the > images where acquired in the range of the detector (=if nothing it > overexposed). Did you acquire all images under the same condition > (excitation power, filter setup, exposure time)? Otherwise it will be > tricky to compare them. > > You can also measure regions by drawing i.e. a circle in the cell and go > for "Analyze" > "Measurements". There are also plugins/macros to automate > this, but a couple of manual steps before are probably go to find the best > way. > > cheers, > Michael > > > > Dear ImageJ, > > I have a question how to use the ImageJ program. I have labeled cells with > > a fluorescence dye. I let the cells in culture about several days. Than I > > made pictures with the fluorescence microscope on day 1, 3 and 5 after > > staining. Now I want to messure and compare the fluorescence intensity > > signal over these days and furthermore I want to examine whether there is > > any weakening. > > So, can you say me how to to this with the ImageJ program? > > > > Thank you. > > > > With kind regards, > > Anette Christ -- Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 [hidden email] (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs |
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