Hello, thanks for watching my videos. Use the selection tool of your interest eg, the rectangular selection tool and select the region of interest. Now run the plugin as shown in the video. All the best with your analysis.
Thank you for this information. I have scanned my slices and I was wondering how should I proceed for analyzing this records. Watching to your tutorial, I would like to ask whether we need to calibrate the measurements for infer the in situ expression of some proteins in the brain by IHQ. May you help me?
Hello, if you have plans to measure the intensity then this discussion might help you www.researchgate.net/post/Does-anyone-have-a-protocol-for-quantifying-IHC-images-in-ImageJ
Hello, I am not sure how to convert %Area into Intensity. This plugin is good for unmixing dyes in images when colors combine subtractively (bright field microscopy with histology stains, watercolors, or printed material with transparent inks). This is the reason It is unsuitable for fluorescence microscopy (fluorophores mix additively) or reflectance imaging that is intensity measurement from %Area. Quantifying staining intensity is only useful when dye uptake is stoichiometric (e.g., the Feulgen reaction for DNA). Unfortunately, immunohistochemical methods are not stoichiometric, making them inappropriate for measuring antigen expression (using intensity).
Glad you found it useful. Here is an interesting discussion on this topic www.researchgate.net/post/Does-anyone-have-a-protocol-for-quantifying-IHC-images-in-ImageJ
Thank you for you helpful video and channel. I am struggling to open up the plugin. After downloading it, it shows a message saying "invalid ou corrupt jarfile". I have tried to open it up in two different laptops but it didn't run. Could you help me please? Thank you.
Thanks for watching my youtube videos. Outlook or gmail does not allow me to send .jar files as it is considered executable file. However, I just found out that this plugin is not supported by the latest ImageJ software update. Please use the older version of the program and run the plugin again. All the best with your analysis.
hello, could you please re-install the plugin again and then re-start imagej. alternately, for windows/mac users copy the plugin and paste it in the "plugins folder" and re-start imagej or drag the plugin into imagej toolbar and then install. let me know if that worked.
Hi!!, Could you help how to calculate the staining intensity only one region of interest?
Hello, thanks for watching my videos. Use the selection tool of your interest eg, the rectangular selection tool and select the region of interest. Now run the plugin as shown in the video. All the best with your analysis.
Thank you for this information. I have scanned my slices and I was wondering how should I proceed for analyzing this records. Watching to your tutorial, I would like to ask whether we need to calibrate the measurements for infer the in situ expression of some proteins in the brain by IHQ. May you help me?
Hello, sure happy to help. Using this plugin only the percentage area is calculated and not the intensity. Are you planning to analyze dab staining?
@@nrttaye4033 yes! my IHQ was performed with DAB staining. I’ve got different brain areas in which my antigen is expressed
Hello, if you have plans to measure the intensity then this discussion might help you www.researchgate.net/post/Does-anyone-have-a-protocol-for-quantifying-IHC-images-in-ImageJ
thank you for you helpful video I need to ask which value will be used for statistical analysis mean or %area. Because I cant see mean in my results
Hello thanks for reaching out. Please use %area for the quantification
Hi, How to convert Area% to intensity?
so only we can draw the graph right?
Hello, I am not sure how to convert %Area into Intensity. This plugin is good for unmixing dyes in images when colors combine subtractively (bright field microscopy with histology stains, watercolors, or printed material with transparent inks). This is the reason It is unsuitable for fluorescence microscopy (fluorophores mix additively) or reflectance imaging that is intensity measurement from %Area. Quantifying staining intensity is only useful when dye uptake is stoichiometric (e.g., the Feulgen reaction for DNA). Unfortunately, immunohistochemical methods are not stoichiometric, making them inappropriate for measuring antigen expression (using intensity).
Thanks for helping it out. Was Struggling from last few days. Could you kindly say how to calculate the staining intensity of IHC ?
Glad you found it useful. Here is an interesting discussion on this topic www.researchgate.net/post/Does-anyone-have-a-protocol-for-quantifying-IHC-images-in-ImageJ
Thanks
Very useful tutorial
Thank you for watching my videos. Gald you found it useful
Thank you for you helpful video and channel. I am struggling to open up the plugin. After downloading it, it shows a message saying "invalid ou corrupt jarfile". I have tried to open it up in two different laptops but it didn't run. Could you help me please? Thank you.
Thanks for watching my youtube videos. Outlook or gmail does not allow me to send .jar files as it is considered executable file. However, I just found out that this plugin is not supported by the latest ImageJ software update. Please use the older version of the program and run the plugin again. All the best with your analysis.
I am trying to install the plugin. But it is not being shown in the software :/
hello, could you please re-install the plugin again and then re-start imagej. alternately, for windows/mac users copy the plugin and paste it in the "plugins folder" and re-start imagej or drag the plugin into imagej toolbar and then install. let me know if that worked.
@nrttaye4033 thank you so much. I copied the file in plugin folder and it worked. Thanks a lot ^^
you are welcome. glad it worked
I tried downloading the plug-in and it’s not working