Open AccessCorrelative microscopy of freeze-dried cells and studies on intracellular calcium stores with imaging secondary ion mass spectrometry (SIMS)
Author(s) -
Subhash Chandra
Publication year - 2019
Publication title -
journal of analytical atomic spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 113
eISSN - 1364-5544
pISSN - 0267-9477
DOI - 10.1039/c9ja00193j
Subject(s) - secondary ion mass spectrometry , mass spectrometry imaging , chemistry , mass spectrometry , analytical chemistry (journal) , ion , microscopy , correlative , calcium , maldi imaging , static secondary ion mass spectrometry , chromatography , matrix assisted laser desorption/ionization , pathology , organic chemistry , linguistics , philosophy , medicine , adsorption , desorption
Secondary ion mass spectrometry (SIMS)-based imaging techniques have become effective tools for studies of elements and molecules in biological samples. In the current work, a correlative microscopy approach was applied to cryogenically prepared fractured freeze-dried cells for organelle-level imaging of chemical composition using SIMS. A CAMECA IMS-3f SIMS ion microscope was used for studying the effect of microtubule-perturbing agents, specifically nocodazole and taxol, on intracellular calcium stores. The perturbation of microtubules in renal epithelial LLC-PK1 cells resulted in significant loss of total calcium in both the nucleus and cytoplasm. In another study, the stable isotope 44 Ca was used for imaging the influx of calcium in resting and stimulated LLC-PK1 cells. SIMS imaging of two calcium isotopes, 44 Ca and 40 Ca, in the same cell revealed the distribution of calcium influx in the 44 Ca image and endogenous calcium in the 40 Ca image. An arginine-vasopressin treatment of cells showed that the Golgi apparatus is sensitive to hormonal stimulation.