Counter staining 1. Incubate cells with 0.1–1 μg/mL Hoechst stain or DAPI (DNA stain) for 1 min. 2. Rinse with PBS. Mounting 1. Mount coverslip with a drop of mounting medium. 2. Seal coverslip with nail polish to prevent drying and movement under microscope. 3. Store in dark at …

An optimized protocol is necessary to achieve consistent, reliable IF results. We have tested common variations in fixation, permeabilization, and antibody concentration for many of our IF-approved antibodies.

Some epitopes may require specific fixation conditions for detection. This is our basic protocol for staining adherent cells in dishes or cells grown on coverslips. Procedure: Rinse cells twice with PBS or HBSS to remove cell culture medium.

Cell Surface Immunofluorescence Staining Protocol Harvest Tissue or Cells: 1. Obtain desired tissue ( e.g . spleen, lymph node, thymus, bone marrow) and prepare a single cell suspension in Cell Staining Buffer (BioLegend Cat. #420201). If using resuspend previously activated cultures in Cell Staining Buffer and proceed to Step 2.

2024年1月23日 · Before staining, warm slides at room temperature for 30 minutes and fix in ice cold acetone for 5 minutes. Air dry for 30 minutes. Wash in PBS; C. Paraffin Sections. Deparaffinize sections in xylene, 2x5min. Hydrate with 100% ethanol, 2x3min. Hydrate with 95% ethanol, 1min. Rinse in distilled water. Follow procedure for pretreatment as required. 2.

Cells are plated at an appropriate density and allowed to attach to the slide or dish (ex. 30,000 cells/chamber in a 8-chamber slide). Cells are usually plated one day prior to staining in order to achieve 60-80% confluency. Note: optimal fixation (both reagent and time) depends on the antigen of interest and must be optimized.

The Thermo ScientificTM NuncTM Lab-TekTM II CC2 Chamber Slide System allows for growth and staining of cells on a single surface. A chamber slide consists of a removable polystyrene media chamber attached to a glass slide treated for adherent cell culture.

2022年2月23日 · This guide can be adapted for several applications, from cell surface immunofluorescence staining to whole-cell imaging, both for immunofluorescence double staining or single-label experiments; use this protocol to achieve high-quality, reproducible imaging in …

Here, we describe preparation of specimens preserved in different types of media and step-by-step methods for both direct and indirect immunofluorescence staining. Keywords: Immunofluorescence, Immunohistochemistry, Immunocytochemistry, Fixation, Antigen Retrieval, Fluorescence, Fluorophore

Immunofluorescence staining or immunocytochemistry is a laboratory technique to detect specific proteins or antigens expression in cells with the aid of selective probes that can be visualized under a fluorescence microscope.