This article describes a method for identifying different types of neural cells derived from peripheral blood. The process involves using fluorescent dye-conjugated antibodies to label specific proteins in these cells, allowing for visualization under a fluorescence microscope.
Take a multi-chamber slide with protein-coated coverslips containing fixed and permeabilized peripheral blood-derived neural cells, including neuronal progenitor cells, astrocytes, and neurons.
Add a blocking solution to block non-specific binding sites.
Incubate with fluorescent dye-conjugated antibodies that bind to specific cellular proteins, enabling the identification of neural cell types.
Wash with buffer to remove unbound antibodies.
Incubate with a fluorescent nuclear dye to stain the nuclei.
Transfer the coverslip to a slide with mounting media. Then, place the prepared slide under a fluorescence microscope equipped with appropriate filters.
Using specific wavelengths of light, excite the fluorescent dyes, causing them to emit light at longer wavelengths.
The emitted fluorescent signals correspond to the labeled cellular proteins, distinctly highlighting different neural cell types and confirming the differentiation of reprogrammed blood cells.
Add a blocking solution of PBS and 5% BSA. Block the cells at room temperature on a rocker plate for one hour. Prepare an appropriate dilution of antibodies in 1% BSA PBS and incubate the cells with the antibodies on a rocker plate for 1.5 hours at room temperature. Wash the cells three times with PBS for five minutes per wash. Incubate the cells with DAPI and mount the coverslips with mounting media for visualization in a microscope.
繁體中文
