What is the first step in the common assay for cytokine detection using flow cytometry?

The first step in the common assay for cytokine detection using flow cytometry is cell collection. This initial process involves obtaining a sample that contains the cells of interest, which may include peripheral blood mononuclear cells, tissue samples, or cell cultures, depending on the specific experimental design. Proper collection of cells is crucial, as it establishes the foundation for the subsequent steps in the assay.

Following cell collection, the cells are typically washed and then may undergo fixation and permeabilization to prepare them for staining with antibodies specific to the cytokines being measured. The fixation step preserves cell structure and cytokine integrity, while permeabilization allows antibodies to access intracellular targets. However, these processes cannot occur until the cells are collected and handled appropriately.

Understanding the sequence of these steps is vital for accurate cytokine detection, as any disruption in the initial collection can affect the quality and viability of the samples, ultimately leading to skewed or irrelevant results in the flow cytometric analysis.