Simultaneous Single-Cell Analysis of Na+, K+, Ca2+, and Mg2+in Neuron-Like PC-12 Cells in a Microfluidic System.
Various intracellular metal ions have closely related functional roles in the nervous system. An excess or deficiency of essential metal ions can contribute to neurodegenerative diseases. Thus, the detection of various metal ions in neurons is important for diagnosing and monitoring these diseases. In particular, single-cell analysis of multiple metal ions allows us to not only understand the cellular heterogeneity and differentiation but also determine the actual relationships among multiple metal ions in each individual cell. Aiming at the low efficient single-cell manipulation and interference of complex biological matrices within cells in the existing method for single-cell metal ion detection, in this manuscript, we present a convenient, sensitive, and reliable method to simultaneously identify and quantify multiple metal ions at the single-cell level using a microfluidic system. Using the combination of on-chip electrophoresis separation and multicolor fluorescence detection, we achieved the simultaneous analysis of Na+, K+, Ca2+, and Mg2+in single PC-12 cells and studied changes in these four metal ions in Aβ25-35-treated PC-12 cells, which is a model of Alzheimer's disease (AD). The data showed that metal ions imbalances in neuron-like cells may be associated with AD induced by Aβ25-35. This method paves the way for multiple metal ion detection in single neuron-like cells, and the results provide insights regarding synergistic function of multiple metal ions in regulation of neurological diseases at the single-cell level.
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Lu
L
Li
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Yuanyuan
Y
Fan
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Qingling
Q
Li
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Renjie
R
Sheng
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Haibin
H
Si
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Juan
J
Fang
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Lili
L
Tong
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Institute of Molecular and Nano Science, Shandong Normal University , Jinan, Shandong 250014, P. R. China.
Bo
0000-0002-8712-7025
B
Tang
D016229:Amyloid beta-Peptides / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000032:analysis* / Q000737:chemistry; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D007477:Ions / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D008274:Magnesium / Q000032:analysis* / Q000737:chemistry; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016716:PC12 Cells; D010446:Peptide Fragments / Q000494:pharmacology; D011188:Potassium / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D059010:Single-Cell Analysis; D012964:Sodium / Q000032:analysis* / Q000097:blood; D013050:Spectrometry, Fluorescence
D016229:Amyloid beta-Peptides; D005456:Fluorescent Dyes; D007477:Ions; D010446:Peptide Fragments; C074636:amyloid beta-protein (25-35); D012964:Sodium; D008274:Magnesium; D011188:Potassium; D002118:Calcium