Immunohistochemical analysis - How? (Jan/30/2007 )
Hi
I am looking at the expression pattern of a tumor marker on a number of tumors, such as breast, colon cancers.
I have 1000s of cancer tissue section slides to read 
.
I don't know how to analysis them .
My boss is unwilling to teach me, even I asked him 
. I had asked him twice already.
I need to finish analyzing all the slides in one month time before my PhD seminar . Or else Minnie is in big trouble.
If you know, please help me.
Thanks in advance.
I am looking at the expression pattern of a tumor marker on a number of tumors, such as breast, colon cancers.
I have 1000s of cancer tissue section slides to read
. I don't know how to analysis them .
My boss is unwilling to teach me, even I asked him
. I had asked him twice already.I need to finish analyzing all the slides in one month time before my PhD seminar
. Or else Minnie is in big trouble. If you know, please help me.
Thanks in advance.
what is the real problem? the myriad of tissue section slides which are already stained, and how to screen them all, or a protocol for IHC staining, or even both?
for each tissue there are working protocols around;
as each stained tissue section needs individual, careful inspection, it is difficult to do an automated screen, so you have to be very busy, or focus, let us say, only on 100 -200 slides;
an unwilling boss means to me, he is not competent in methods, or in having time for his students, and, therefore, he is not competent to supervise a doctoral thesis; you should tell this to the next-generation students in your lab or institute...
Its tricky to analyze immuno staining without help. One advise would be to take a particular marker and check in literature to know how is the normal expression and look at pic in papers. Take slides stained to indentify these markers. Look at the control tissue, all regions carefully, spend some time with it. Then look at the actual slides and compare them. Hopefully you should see some differences in expression pattern.
for each tissue there are working protocols around;
as each stained tissue section needs individual, careful inspection, it is difficult to do an automated screen, so you have to be very busy, or focus, let us say, only on 100 -200 slides;
Hi Bearer,
Thank you for replying my post.
All the tissue sections are stained. I just don't know how to analyze them properly.
I don't want to waste my time (only have 6-9 months left to do experiements) to analyze them, then later find out that I have done them wrong and have to re-analyze them again and again.
It is OK if I have only less than 50 slides, but I have ~2000 slides...analyze them takes time and re-analyse them also takes time.
I asked my boss again, he told me just analyze it myself and figure it out myself.
I think if he is willing to teach me, it would save me heap of time to analyze it.
He told me that he had ordered a histology reference book to me, but Minnie already got one at home.
His previous student already told me that he is a OK supervisor.
Thank you scolix for the advise.
The problem is the marker is very unique and nobody is studying on it...therefore no reference in the literature.
I think you are right that I may need to compare the control tissue and the actual slides and try to find the differences expression pattern between them.
Thank you
And one more thing,
if you think there might be a diffeence in the expression pattern, ask one of your colleagues or friends to look at the slides (just one control and one of the actual one) and ask them if there is any difference. Ofcourse, they should have no idea which slide is control or not. And if they see an obvious difference, then you might actually have a pattern. This is only a way if your supervisor is not willing to help you.
Good Luck !!!
Thank you very much. Scolix