I need a method for the quantitative evaluation of histological slides obtained after microtome sectioning of the rat prostates. The experiment was performed to assess the effect of herbal extracts on Benign Prostatic Hyperplasia induced by testosterone treatment. The qualitative evaluation is possible in the sense that luminal evolutions, stromal compartment and epithelial cells morphology can be seen. But as far as quantitative evaluation is concerned, I have found no parameter which can be measured using these slides?
Parameters such as luminal volume, stromal area or the involutions etc. need to be measured quantitatively for a better comparison of treated and control groups. I am attaching a photograph of slide for exemplification.
Thanks in advance for your support.
I do not have any exerience in this area of research but could you use computer stimulation as this researches seem to suggest. Check
www.pnas.org/cgi/doi/10.1073/pnas.1217982109
Kiswowski et al. 2011, Cancer Res;71 (10) May 15,2011
Kiswowski and Jackson, Cancer Res Online ISSN: 1538-7445, J. Cancer Res ISSN 0099-7013, Am J Cancer ISSN: 0099-7374. I hope this is helpful.
Thanks Mary. Hope to get something out of your suggestions.
Best Regards
immunohistochemical staining of the markers and then quantification of this stain by measuring the optical denisty of 10 fields from each group ( then taking the average) give a quantification for the severity of the cancer
i mean markers of the cancer, in my work i quantify inflammation by measuring inflammatory markers like tnf alpha, PGE2 ........
@ Doaa Korany... I have the slides of the prostate finally as seen in the image attached. What can be done with them for quantification because I never performed immunohistochemical staining. and also didnt measure inflammatory markers like you.
James Denegre at Jackson Laboratories is an expert at this. [email protected]
You will neet to submit the glass and a computerized scan can be done to thereby run a programmed quantification. It costs about $20 per slide but the results are good. I reccomend James for expertise such as this. He is resourceful. Best, Bob
HI, I am not sure if I fully understood the question. Do you have in your university a microscope with a software called "Stereoinvestigator"? Could you use the cavalieri method? Some publications use the cavalieri method for area analysis in cancer vs. the normal counterpart. Not sure if this is the case.
@Francesco Marino. Thanks for your response. We don't have "Stereoinvestigator" with us. I am not sure as to we can use cavalieri method or not because these are the slides of a hyperplasic prostate gland and not a cancerous growth. Just need a measure to quantify the increase in luminal volume and area, stromal area, the connective tissue in between the cells, the luminal involutions etc. I am stuck here. Can't find a way out because trinocular microscope has already been used to get the photographs as seen in the slides. Now quantitative evaluation has become a problem as nobody has done it before for prostate gland as per my literature reviews. I also tried to use the quantitative microscopic techniques using the eyepiece and stage micrometer but couldn't get any fruitful results to correlate with the mentioned parameters.
Regards
Cavalieri method is usually used to estimate the volume of the tissue your are investigating. I can't see why you can't use it, I mean it doesn't matter if you have a cancerous tissue or hyperplasia you are measuring the same thing a volume. With the cavalieri method you can use a rectangular lattice of points on the area of interest and then counting the number of points in the lattice that lie within the area of interest.
You can use Image J and download a plugin called Volumest ( this is an open source software). With the plugin you should be able to have a try using the volume estimator with the cavalieri method.
@ Francesco Elia Marino.......I had gone through the method but I couldn't find the practical approach as to how to perform the analysis. I am attaching the pdf which i referred? Pls help and if you can apply the method to the slides in the figure attached with the question, pls guide me through the process. I can send you other slides too for the same purpose. Your help will be highly appreciated.
Thanks and Best Regards
The Cavalieri Technique you sent me is a general formula and maths calculation for geometry and other application. Now you should have a look at the Stereology application of the technique. For example this file could help you. But you need to have a program for microscopy analysis in order to calculate area and/or volume.
Regards,
Francesco
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