![]() ![]() While perfect correlation gives a value of 1, perfect exclusion does not give a value of -1. However, this is not the case for images. A value of 1 represents perfect correlation -1 represents perfect exclusion and zero represents random localisation. In many forms of correlation analysis the values for Pearson’s will range from 1 to -1. This is a popular method of quantifying correlation in many fields of research from psychology to economics. Zero-zero pixels are not included in this calculation. This is the Pearson’s correlation coefficient. Select max intensity from the pulldown list.Flatten z-stacks by going Image->Z-Project and selecting the range of images (typically the first x images are one color and then the last x images are the next color).Open image and change to 8-bit (Image->Type->8 bit).Assess colocalisation by Plugins->Analyze->Colocalisation Finder.Select each image and change to 8-bit greyscale by Image->type->8bit.Select File and split by Image->color->Split.Select File and create composite by Image->color>create composite.Open an experiment file in ImageJ by File->Open->Select File.Prior to performing colocalisation analysis save cell images as experiment files. ![]() If you need any more information, I am happy to provide it. What software packages and/or plugins have you tried?.I just want to get a bit more proficient in ImageJ before tackling other cell lines. Is there any way to automate/batch processing this? The reason is because I have a quite several images that needs to undergo the same image j processing for different cell lines. I seemed to get lost on step three, because they are three channels. My first query is that is the protocol listed above appropriate.Īre there advise to improve the protocol? Results of measured intensity of fluorescence along the drawn line Go to Analyze => "Histogram A window should appear with the graph and NOTE: Be aware on which channel you draw the line, even if both channelsĪre shown, only 1 is measured (To change channel, scroll left or right, title ofĬhannel should be displayed at top of the window,) A yellow line should appear on stacked image To measure fluorescent intensity: draw line (or other shape chosen from Window appears with Colour and possibility to tick boxes of both channels.įirst tick box of Channel 1 and click More to assign appropriate colourĬhange Colour to Composite to view stack with both colours. To question asking: Convert to multi-channelĭisplay mode: change from Composite to Colour and click OK To assign the right colour to each channel, click on "Image => “Colour” =>Ĭhange Composite to Colour. Right to switch between channels both channels are shown in the same Now both channels appear in the same window (you need to scroll left and Using ImageJ to Measure Intensity of FluorescenceĢ.Go to "Image => "Stacks => “Images to Stack” Name stack, e.g. I have been provided the following protocol: What information are you interested in getting from this image? They are breast cancer cell lines called BT549 What is the image about? Provide some background and/or a description of the image. My task is to determine if the glycosidase inhibitor treatment that I am asked to evaluate has indeed altered the cellular glycosylation by examining the lectin florescence stating of the cancer cells for those treated with my compound and compared to the untreated and the positive control (deoxynojirimycin). Scan_Plate_R_p00_0_B02f00d1.TIF is the green image Analysis goals Scan_Plate_R_p00_0_B02f00d0.TIF is blue image I have been told to use the * “scan _Plate_R…” for my analysis. Images labelled “scan _Plate_R…” split the green (lectin) and blue(DAPI, nuclear stain). Have green (lectin) and blue (DAPI, nuclear stain) overlaid (merged) Share a minimal working example of your macro code.Upload an original image file here directly or share via a link to a file-sharing site (such as Dropbox) – (make sure however that you are allowed to share the image data publicly under the conditions of this forum). ![]()
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