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Immunohistochemistry Forum Immunohistochemistry Forum. Post and discuss questions on immunohistochemistry methods including paraffin, frozen, or free floating tissue sections.


Immunohistochemistry Quantification

Immunohistochemistry Quantification - Immunohistochemistry Forum

Immunohistochemistry Quantification - Immunohistochemistry Forum. Post and discuss questions on immunohistochemistry methods including paraffin, frozen, or free floating tissue sections.


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  #1  
Old 11-26-2007, 08:24 AM
Pipette Filler
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Question Immunohistochemistry Quantification



Hello guys,

I am having problems quantifying of my immunohistochemistry results.

I am getting significance when I see the results, however with Image J it shows no difference.


Is this due to the fact that Image J is not sensitive enough?

What do you guys think?

thanks in advance

Last edited by admin; 11-26-2007 at 08:28 AM.
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Old 12-04-2008, 12:34 AM
Pipette Filler
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Default Re: Immunohistochemistry Quantification

Hello:
What are you quantifying? cells,areas,nucleus... fluorescence or chromogen? I have used ImageJ ocasionally, normally we use the AnalySIS program but they are similar. The problem could be the tresholds, the ROI selecction or for example the size of the cells if you are quantifying areas (not the same in all the samples/zones), the quality of the images...
I donīt know if i can help you...but i need more information to try...
My opinion is that if you see evident diferences between samples under the microscope ,the program should detect them if all the settings and the images are ok.
Best regards
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Old 04-23-2009, 10:25 PM
Pipette Filler
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Default Re: Immunohistochemistry Quantification

I assume that you are attempting to quantify the expression of a nuclear, membranous, or cytoplasmic marker. Please provide more information, including which marker you are studying, which area you are evaluating (e.g., an area of interest versus the whole slide), and what quantitative threshhold you are employing.
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Old 05-30-2009, 03:24 PM
Pipette Filler
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Default Re: Immunohistochemistry Quantification

I have the same problem, as I need some help on how to use image J to quantify my immunohistochemistry results as I need to quantify the intensity of the florescence staining in my slides?!!
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Old 10-21-2010, 12:54 PM
Pipette Filler
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Question Re: Immunohistochemistry Quantification

Hello all!
Im having problems with cellular quantification. Im looking for differences on protein quantities between sister blastomeres but Im not sure how to stablish a background to compare relative ammounts of proteins... Does anyone have a clue?
Best regads
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Old 11-02-2010, 08:58 PM
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Default Re: Immunohistochemistry Quantification

Pipette_man - what do you mean when you say you are "getting significance when [you] see the results" - do you mean they just appear significant to you visually? If so, I wouldn't put much weight in that. Any chance you can post the images or the quantified data? That would make things easier to assess. Also, along the lines with what radam said, please include what method you are using to quantify the intensity so we know how you are performing the analysis.

Also, (and I tell everyone this over and over and over to make sure this isn't the problem) but make sure you're not having the microscope software auto-adjust the exposure levels for each image. You need to make sure the exposures are the same to be able to quantitatively compare any images.
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