Variation of increases in free cytosolic calcium ion induced by prostaglandin E2 in mouse osteoblast clone, MC3T3-E1—Single cell assay using microspectro-fluorometry

The new, highly fluorescent, calcium-sensitive dye, fura-2, can be loaded nondisruptively into intact cells by means of its permeant ester and used to measure the free calcium ion concentration in individual cells. For fura-2 to signal cytosolic calcium, it must be distributed homogeneously and exclusively throughout the cytoplasmic space. However, microscopic examination of bovine aortic endothelial cells loaded with fura-2 by exposure to its permeant ester reveals fluorescence associated with discrete intracellular structures rather than the homogeneous distribution expected for a cytosolic stain. Simultaneous labeling of bovine aortic endothelial cells with fura-2 and rhodamine 123 (a mitochondrial fluorescent vital stain) identifies these structures as mitochondria. Subcellular dye localizations are not observed when the cells are loaded with other putative cytosolic stains that gain access to the cytosol by means of a membrane permeant ester. Both carboxyfluorescein and indo-1 (another member of the family of second generation calcium indicators) stain the cytoplasm diffusely. It is suggested that fura-2 fluorescence accumulates in certain cells in association with mitochondria. It is important to assess the intracellular distribution of fura-2 when this indicator is used to measure the free cytosolic calcium ion concentration.

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Simultaneous Measurements of Cytosolic Calcium Ion and pH in Cultured Superior Cervical Ganglion Cells of Rat

Frontiers in Arterial Chemoreception - Advances in Experimental Medicine and Biology ◽

10.1007/978-1-4615-5891-0_32 ◽

1996 ◽

pp. 221-226

Author(s):

K. Yoshizaki ◽

K. Ohtomo ◽

K. Fukuhara ◽

T. Kawaguchi ◽

J. Kawaguchi

Keyword(s):

Superior Cervical Ganglion ◽

Calcium Ion ◽

Ganglion Cells ◽

Cytosolic Calcium ◽

Simultaneous Measurements ◽

Cervical Ganglion

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Suppressive effects of low-power laser on cytosolic calcium ion changes elicited by bradykinin in cultured murine dorsal root ganglion neurons

PAIN RESEARCH ◽

10.11154/pain.20.127 ◽

2005 ◽

Vol 20 (3) ◽

pp. 127-133 ◽

Cited By ~ 1

Author(s):

Kentaro Yoda ◽

Youichi Kuzuoka ◽

Junko Ohtani

Keyword(s):

Dorsal Root Ganglion ◽

Low Power ◽

Calcium Ion ◽

Dorsal Root ◽

Cytosolic Calcium ◽

Dorsal Root Ganglion Neurons ◽

Power Laser ◽

Ganglion Neurons

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Cell elastimetry in the detection of antineutrophil antibodies

Blood ◽

10.1182/blood.v57.1.22.bloodjournal57122 ◽

1981 ◽

Vol 57 (1) ◽

pp. 22-24

Author(s):

ME Miller ◽

LA Boxer ◽

EJ Kawaoka ◽

WA Border

Keyword(s):

Single Cell ◽

Immune Complexes ◽

Negative Pressure ◽

Direct Visualization ◽

Cell Assay ◽

Membrane Rigidity ◽

Blind Fashion

Cell elastimetry has been applied to the measurement of antineutrophil antibodies. This technique measures, under direct visualization, the negative pressure required of aspirate PMNs into small-pored pipettes. Two groups of studies were carried out: (A) In the first group of studies, normal PMNs were incubated with 1 of 8 known antineutrophil serums. Each serum significantly decreased membrane deformability-- i.e., cells became more rigid. The study was conducted in an entirely blind fashion. Randomly coded serums from patients and controls were studied for deformability by observers unaware of the code. (B) In the second group of studies, sera containing immune complexes were incubated with normal PMNs. No significant effects were noted upon deformability. As a single cell assay that partially reflects membrane rigidity, elastimetry may, therefore, have potential in the further characterization of mechanisms by which such antineutrophil antibodies compromise neutrophil functions.

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