Calcein AM/PI Assay: A Step-by-Step Guide for Cell Viability Assessment

Calcein AM/PI Assay

This guide provides a step-by-step procedure for performing a Calcein AM/PI assay to assess cell viability. This method utilizes the fluorescent dyes Calcein AM and Propidium Iodide (PI) to distinguish between live and dead cells.

Preparation

1. Prepare the Calcein AM/PI working solution (manufactured by Beyotime, China) freshly. Follow the manufacturer's instructions carefully and mix well before use. Avoid freezing any unused working solution for future use.

Sample Preparation

2. Remove the spent media from the 48-well plate or remove the coverslip of the eCHIP. Carefully wash the plate or eCHIP once with PBS.

3. Add an appropriate amount of the Calcein AM/PI working solution. Ensure that the solution completely covers the hydrogel-cell complex.

Incubation

4. Place the plate or eCHIP in a 37℃ cell incubator for 30 minutes.

Microscopy Analysis

5. After incubation, examine the sample under a fluorescence microscope (Nikon, Japan).

Important Notes:

• The entire procedure should be protected from light.
• The Calcein AM/PI assay is a sensitive method that can be used to assess the viability of a wide range of cell types.