Affiliation:
1. Department of Biomedical Engineering Cornell SIMS Laboratory Cornell University Ithaca New York
Abstract
AbstractA secondary ion mass spectrometry (SIMS)‐based isotopic imaging technique of ion microscopy was used for observing calcium influx in single renal epithelial LLC‐PK1 cells. The CAMECA IMS‐3f SIMS instrument, used in the study, is capable of producing isotopic images of single cells at 500 nm spatial resolution. Due to the high‐vacuum requirements of the instrument the cells were prepared cryogenically with a freeze‐fracture method and frozen freeze‐dried cells were used for SIMS analysis. The influx of calcium was imaged directly by exposure of cells to 44Ca stable isotope in the extracellular buffer for 10 min. The 44Ca influx was measured at mass 44 and the distribution of endogenous calcium at mass 40 (40Ca) in the same cell. A direct comparison of interphase cells to cells undergoing division revealed that calcium influx is restricted in metaphase and post‐metaphase stages of cell division. This restriction is lifted in late cytokinesis. The net influx of 44Ca in 10 min was approximately half under calcium influx restriction in comparison to interphase cells. Under calcium influx restriction the 44Ca concentration was the same between the mitotic chromosome and the cytoplasm. These observations indicate that the endoplasmic reticulum (ER) calcium uptake is compromised under calcium influx restriction in cells undergoing division.
Funder
National Institutes of Health
Subject
Histology,Pathology and Forensic Medicine
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献