A new method of preparing human whole brain sections for in vitro receptor autoradiography.
In vitro receptor autoradiography is a widely used technique for determining the distribution of radioligand binding sites. By using this technique it is possible to investigate alterations in receptor number and affinity caused by trauma or a disease state. To date, however, the largest sections prepared for in vitro autoradiography have been from single human hemispheres, with the adjacent hemisphere being used for neuropathological investigations. Therefore, a method for cutting large cryosections of human whole brain tissue is described. Whole brains obtained less than 2 days postmortem were frozen at -80 degrees C. 1.5-2 cm coronal slices were cut from the brain and embedded and frozen in a carboxymethylcellulose solution. Sections 40 microm in size were sliced from the frozen block at -16 degrees C in a whole body cryostat. The sections were lifted by means of a nylon membrane backing material and subsequently incubated with tritiated ligand to produce autoradiograms of each whole brain coronal section. [(3)H]paroxetine was used in the present study as an example.
Miller
J K
JK
The School of Pharmacy, London University, UK.
Bowery
N G
NG
Tulloch
I F
IF
D000368:Aged; D000369:Aged, 80 and over; D001345:Autoradiography / Q000379:methods*; D001923:Brain Chemistry; D006801:Humans; D008297:Male; D008867:Microtomy / Q000379:methods*; D008875:Middle Aged; D017374:Paroxetine; D011869:Radioligand Assay; D017367:Selective Serotonin Reuptake Inhibitors; D016610:Tissue Embedding / Q000379:methods*; D014316:Tritium
D017374:Paroxetine; D017367:Selective Serotonin Reuptake Inhibitors; D014316:Tritium