20181106: IgG and IgM primary culture Schwann cell ICC in Zika-GBS patients (Willison cohort)

Primary culture Schwann cell IgG and IgM ICC is performed to test Zika-GBS patients staining pattern.

Samples: 11 sera from Zika-GBS (Willison cohort), and 2 negative controls.

Protocol:

1. Primary culture Schwann cells  grown in coverslips were fixed with PFA 4 % 10 min,  blocked with Goat 5% in PBS 1h, and frozen at -80ºC.
2. Thaw these cells with a little Goat 5% in PBS
3. Patient’s sera 1/40 for IgM and 1/100 for IgG 1h at RT
4. Wash 3 times with PBS.
5. Secondary Ab GAH 488 IgG or IgM (1/1000) 1h at RT
6. Wash 3 times with PBS.
7. Mount with Vectashield mounting medium.

Results: