PFA fixation assay of cells
PFA fixation assay of cells
When pure formaldehyde is dissolved in aqueous solution, it automatically polymerizes into long-chain paraformaldehyde. Its length varies and the polymerization and depolymerization reactions take place simultaneously in water. Formaldehyde is readily soluble in lipids and can therefore cross cell membranes into cells. When cells are immobilized with paraformaldehyde, it chemically cross-links the free amino groups within the cells. When different molecules are cross-linked, a mesh structure is formed, connecting the various structural components of the cell.
Operation method
levitation
Materials and Instruments
Cell Suspensions Move 1. Aspirate 10 μl of cell suspension at a concentration of 2×107 cells/ml onto a poly-L-lysine-coated slide. Place in a humidifying box and allow the cells to stand for 30 min. For more product details, please visit Aladdin Scientific website.
PFA PBS
Slides Humidifying cassettes Glass staining trays
2. Place the slide in a slide holder and immerse it in a staining glass dish containing 4% PFA fixative and fix it for 20 min at room temperature.3. Transfer the slide holder to another plate with 3 x PBS and leave for 2 min (this step terminates PFA fixation).
4. Transfer the slide holder to a new dish containing 1×PBS and leave it for 2 min. change 1×PBS and leave it for another 2 min.
5. Add 50%, 70%, 95% and 100% ethanol to the tray and leave it in each solution for 5 minutes for dehydration.6. Dry slides completely in air, place in slide box with desiccant and store at -70°C (for several weeks).