[Histonet] maximum thickness brain in slide

[Colleen Forster]

Even 30um is too thick for the routine IHC on glass. You will never
penetrate the whole thickness for staining.

Colleen Forster HT(ASCP)QIHC

On Fri, Aug 4, 2023 at 2:05 PM Alonso Martínez Canabal <
acanabal at ciencias.unam.mx> wrote:

> Hi, thank you for your reply.
>     I cannot do my usual free floating staining because this tissue is
> very brittle. So, I was thinking to do the staining in slide at 30 microns?
> Thank you
>
> El vie, 4 ago 2023 a las 12:29, Colleen Forster (<cforster at umn.edu>)
> escribió:
>
>> Alonso,
>>
>> For sections that thick you would collect them as you always have, stain
>> them as floating sections and then mount onto a glass slide. The routine
>> standard methods for IHC will never penetrate a  50um thick section.
>>
>> Colleen Forster HT(ASCP)QIHC
>>
>>
>>
>> On Fri, Aug 4, 2023 at 1:17 PM Alonso Martínez Canabal via Histonet <
>> histonet at lists.utsouthwestern.edu> wrote:
>>
>>> Good afternoon Histoneters,
>>>        I have been doing IHC and IF during decades in rodent brain
>>> tissue,
>>> always in free floating. Now, I would like to do that sections placed
>>> over
>>> slides and perform the whole procedure over the slide. I am interested in
>>> using thick sections, usually I used 50micrometers, what is the maximum
>>> thickness that I could use in slide to guarantee the correct penetration
>>> of
>>> the antibodies and the ABC complex. Before the IHC I always use graded
>>> alcohols and xilene, and antigen retrieval with citrate buffer.
>>>       Thank you!
>>>
>>> --
>>> Dr. Alonso Martínez Canabal PhD
>>> Profesor Asociado "C"
>>> Departamento de Biología Celular, Facultad de Ciencias, UNAM
>>> Investigador Nacional "I"
>>> 56224833
>>> _______________________________________________
>>> Histonet mailing list
>>> Histonet at lists.utsouthwestern.edu
>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>
>>
>> --
>> Colleen Forster HT(ASCP)QIHC
>> BLS Histology and IHC Laboratory
>> Jackson Hall, Room 2-155
>> 321 Church St. SE
>> Minneapolis, MN 55455
>> 612-626-1930
>>
>>
>>
>>
>>
>>
>>
>
> --
> Dr. Alonso Martínez Canabal PhD
> Profesor Asociado "C"
> Departamento de Biología Celular, Facultad de Ciencias, UNAM
> Investigador Nacional "I"
> 56224833
>
>

-- 
Colleen Forster HT(ASCP)QIHC
BLS Histology and IHC Laboratory
Jackson Hall, Room 2-155
321 Church St. SE
Minneapolis, MN 55455
612-626-1930