Optimized Staining Methods of Intracellular Calcium for RAW264.7 Cells

¹ Biomechanics Lab, Department of Mechanics, School of Aerospace Engineering, Beijing Institute of Technology, Beijing 100081, P. R. China
² Institute of Artificial Intelligence in Sports, Capital University of Physical Education and Sports, Beijing 100191, P.R. China

^* Corresponding author: huobo@cupes.edu.cn

Abstract

Monitoring intracellular calcium ion levels accurately and efficiently is crucial for gaining a profound understanding of cellular functionality. Inappropriate staining conditions may result in deviations in fluorescent signals, which affects the accuracy and repeatability of experimental outcomes. To measure the concentrations change of Ca²⁺ accurately, immunofluorescent staining of intracellular calcium is an essential process. In order to solve the previous dyeing problem of our research group, we designed and performed an experimental scheme. The results indicated that a staining duration of 30 minutes with a dye concentration at a 1/1000 dilution ratio consistently yielded the most reliable and stable staining outcomes. The results of this study have established the foundation of the future experiments and may be further helpful to provide a valuable foundation for the development of therapeutic strategies targeting bone diseases.