Nondestructive quantum dot-based intracellular serotonin imaging in intact cells
Section snippets
Cells and serotonin uptake
Human choriocarcinoma cells (JAR) were cultured in RPMI 1640 medium (30041, JRS, Inc.). Cultures were supplemented with 10% v/v fetal bovine serum (FBS; 16000-044, Gibco), 60 μg/ml penicillin, and 100 μg/ml streptomycin (P0781, Sigma) and maintained in 25 cm2 cell culture flasks (Nunclon) at 37 °C in a 5% CO2 atmosphere. After 48 h in culture, cells were used for uptake experiments. The culture medium was removed and cells were washed briefly in 1× phosphate-buffered saline (PBS) and preincubated
Results and discussion
Native autofluorescence of JAR cells can be seen in Fig. 2a. Cells were excited at 488 nm (in the absence of Qdot-antibody conjugates and serotonin) and autofluorescence was detected at 523 nm. Numerous intracellular fluorescent molecules that emit light at specified wavelengths have been reported as biomarkers to distinguish normal cells from neoplastic cells. These biomarkers include a large number of proteins, coenzymes, and micronutrients. Among the native intracellular fluorescent molecules,
Conclusion
This study provides a sophisticated approach that directly detects intracellular components such as serotonin granules in intact cells. The serotonin neurotransmitter is directly visualized via a nondestructive multicolor immunofluorescence assay. A set of cellular images as a function of wavelength is obtained without much cellular destruction. This multicolor imaging technique clearly does away with aggregation issues that emerge from the use of quantum dots as probes for intracellular
Acknowledgments
This work was supported by a Korea Science and Engineering Foundation (KOSEF) grant funded by the Korean Ministry of Science and Technology (MOST) under contract M10640040002-08N4004-00211 and M10644080003-08N4408-00310.
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