A new antigen retrieval technique for human brain tissue.
Immunohistochemical staining of tissues is a powerful tool used to delineate the presence or absence of an antigen. During the last 30 years, antigen visualization in human brain tissue has been significantly limited by the masking effect of fixatives. In the present study, we have used a new method for antigen retrieval in formalin-fixed human brain tissue and examined the effectiveness of this protocol to reveal masked antigens in tissues with both short and long formalin fixation times. This new method, which is based on the use of citraconic acid, has not been previously utilized in brain tissue although it has been employed in various other tissues such as tonsil, ovary, skin, lymph node, stomach, breast, colon, lung and thymus. Thus, we reported here a novel method to carry out immunohistochemical studies in free-floating human brain sections. Since fixation of brain tissue specimens in formaldehyde is a commonly method used in brain banks, this new antigen retrieval method could facilitate immunohistochemical studies of brains with prolonged formalin fixation times.
Alelú-Paz
Raúl
R
Departamento de Anatomía, Histología y Neurociencia, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain.
Iturrieta-Zuazo
Ignacio
I
Byne
William
W
Haroutunian
Vahram
V
García-Villanueva
Mercedes
M
Rábano
Alberto
A
García-Amado
María
M
Prensa
Lucía
L
Giménez-Amaya
José Manuel
JM
D000941:Antigens / Q000032:analysis*; D001923:Brain Chemistry / Q000276:immunology*; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D005650:Fumarates; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods* / Q000592:standards; D008297:Male; D008298:Maleates; D008722:Methods; D016707:Tissue Fixation
D000941:Antigens; D005404:Fixatives; D005650:Fumarates; D008298:Maleates; C073341:citraconic acid; D005557:Formaldehyde