Tracking DNA sensor and inflammasome complexes in cells using confocal microscopy

2019 ◽  
pp. 253-267
Author(s):  
Brendan Antiochos
Keyword(s):  
Confocal Microscopy ◽  
Dna Sensor ◽  
In Cells

SU-F-T-665: Confocal Microscopy Imaging of Cell Cycle Distribution in Cells Treated with Pegylated Gold Nanoshells

2016 ◽  
Vol 43 (6Part23) ◽  
pp. 3617-3617
Author(s):  
D Sadetaporn ◽  
D Flint ◽  
C McFadden ◽  
A Asaithamby ◽  
G Sawakuchi
Keyword(s):  
Cell Cycle ◽  
Confocal Microscopy ◽  
Gold Nanoshells ◽  
Cell Cycle Distribution ◽  
Microscopy Imaging ◽  
Confocal Microscopy Imaging ◽  
In Cells

scholarly journals Primitive organization of cytosolic Ca(2+) signals in hepatocytes from the little skate Raja erinacea

1999 ◽  
Vol 202 (22) ◽  
pp. 3049-3056
Author(s):  
M.H. Nathanson ◽  
A.F. O'Neill ◽  
A.D. Burgstahler
Keyword(s):  
Confocal Microscopy ◽  
Subcellular Distribution ◽  
Immunoblot Analysis ◽  
Apical Region ◽  
Type I ◽  
Raja Erinacea ◽  
Inositol 1 ◽  
Cell Functions ◽  
Confocal Immunofluorescence ◽  
In Cells

Cytosolic Ca(2+) (Ca(i)(2+)) signals begin as polarized, inositol 1, 4,5-trisphosphate (InsP3)-mediated Ca(i)(2+) waves in mammalian epithelia, and this signaling pattern directs secretion together with other cell functions. To investigate whether Ca(i)(2+) signaling is similarly organized in elasmobranch epithelia, we examined Ca(i)(2+) signaling patterns and InsP3 receptor (InsP3R) expression in hepatocytes isolated from the little skate, Raja erinacea. Ca(i)(2+) signaling was examined by confocal microscopy, InsP3R expression by immunoblot, and the subcellular distribution of InsP3Rs by immunochemistry. ATP induced a rapid increase in Ca(i)(2+) in skate hepatocytes, as it does in mammalian hepatocytes. Unlike in mammalian hepatocytes, however, the Ca(i)(2+) increase in skate hepatocytes began randomly throughout the cell rather than in the apical region. In cells loaded with heparin ATP-induced Ca(i)(2+) signals were inhibited, but de-N-sulfated heparin was not inhibitory, suggesting that the increases in Ca(i)(2+) were mediated by InsP3. Immunoblot analysis showed that the type I but not the types II or III InsP3R was expressed in skate liver. Confocal immunofluorescence revealed that the InsP3R was distributed throughout the hepatocyte, rather than concentrated apically as in mammalian epithelia. These findings demonstrate that ATP-induced Ca(i)(2+) signals are mediated by InsP3 in skate hepatocytes, as they are in mammalian hepatocytes. However, in skate hepatocytes Ca(i)(2+) signals begin at loci throughout the cell rather than as an organized apical-to-basal Ca(i)(2+) wave, which is probably because the InsP3R is distributed throughout these cells. This primitive organization of Ca(i)(2+) signaling may in part explain the observation that Ca(2+)-mediated events such as secretion occur much less efficiently in elasmobranchs than in mammals.

Cytoplasmic Tubules in Cells Infected with Laryngotracheitis Virus

1969 ◽  
Vol 27 ◽  
pp. 376-377
Author(s):  
A. M. Watrach
Keyword(s):  
Tissue Culture ◽  
Small Proportion ◽  
Chicken Embryo ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Morphologic Characteristics ◽  
Infectious Laryngotracheitis ◽  
Laryngotracheitis Virus ◽  
In Cells

During a study of the development of infectious laryngotracheitis (LT) virus in tissue culture cells, unusual tubular formations were found in the cytoplasm of a small proportion of the affected cells. It is the purpose of this report to describe the morphologic characteristics of the tubules and to discuss their possible association with the development of virus.The source and maintenance of the strain of LT virus have been described. Prior to this study, the virus was passed several times in chicken embryo kidney (CEK) tissue culture cells.

Effect of Some Metabolic Inhibitors on Vinblastine-Induced Ribosomal-Granular Material Complexes

1971 ◽  
Vol 29 ◽  
pp. 548-549
Author(s):  
Awtar Krishan ◽  
Dora Hsu
Keyword(s):  
Granular Material ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Actinomycin D ◽  
Metabolic Inhibitors ◽  
Protein And Rna Synthesis ◽  
Apparent Reduction ◽  
Plant Alkaloids ◽  
In Cells

Cells exposed to antitumor plant alkaloids, vinblastine and vincristine sulfate have large proteinacious crystals and complexes of ribosomes, helical polyribosomes and electron-dense granular material (ribosomal complexes) in their cytoplasm, Binding of H3-colchicine by the in vivo crystals shows that they contain microtubular proteins. Association of ribosomal complexes with the crystals suggests that these structures may be interrelated.In the present study cultured human leukemic lymphoblasts (CCRF-CEM), were incubated with protein and RNA-synthesis inhibitors, p. fluorophenylalanine, puromycin, cycloheximide or actinomycin-D before the addition of crystal-inducing doses of vinblastine to the culture medium. None of these compounds could completely prevent the formation of the ribosomal complexes or the crystals. However, in cells pre-incubated with puromycin, cycloheximide, or actinomycin-D, a reduction in the number and size of the ribosomal complexes was seen. Large helical polyribosomes were absent in the ribosomal complexes of cells treated with puromycin, while in cells exposed to cycloheximide, there was an apparent reduction in the number of ribosomes associated with the ribosomal complexes (Fig. 2).

Two-photon excitation microscopy in cellular biophysics

1995 ◽  
Vol 53 ◽  
pp. 62-63
Author(s):  
David W. Piston ◽  
Brian D. Bennett ◽  
Robert G. Summers
Keyword(s):  
Confocal Microscopy ◽  
Laser Beam ◽  
Duty Cycle ◽  
Excited Electronic State ◽  
Input Power ◽  
Two Photon ◽  
Simultaneous Absorption ◽  
Photon Excitation ◽  
Very High ◽  
Two Photon Excitation

Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. Two-photon excitation arises from the simultaneous absorption of two photons in a single quantitized event whose probability is proportional to the square of the instantaneous intensity. For example, two red photons can cause the transition to an excited electronic state normally reached by absorption in the ultraviolet. In practice, two-photon excitation is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10-5 maintains the average input power on the order of 10 mW, only slightly greater than the power normally used in confocal microscopy.

Measurement of spontaneous neuronal membrane activity by time lapse confocal microscopy

1995 ◽  
Vol 53 ◽  
pp. 972-973
Author(s):  
R H. Selinfreund ◽  
A. H. Cornell-Bell
Keyword(s):  
Membrane Potential ◽  
Confocal Microscopy ◽  
Patch Clamp ◽  
Electrical Activity ◽  
Time Lapse ◽  
Neuronal Firing ◽  
Neuronal Membrane ◽  
Pituitary Cells ◽  
Patch Clamp Recording ◽  
Spontaneous Electrical Activity

Cellular electrophysiological properties are normally monitored by standard patch clamp techniques . The combination of membrane potential dyes with time-lapse laser confocal microscopy provides a more direct, least destructive rapid method for monitoring changes in neuronal electrical activity. Using membrane potential dyes we found that spontaneous action potential firing can be detected using time-lapse confocal microscopy. Initially, patch clamp recording techniques were used to verify spontaneous electrical activity in GH4\C1 pituitary cells. It was found that serum depleted cells had reduced spontaneous electrical activity. Brief exposure to the serum derived growth factor, IGF-1, reconstituted electrical activity. We have examined the possibility of developing a rapid fluorescent assay to measure neuronal activity using membrane potential dyes. This neuronal regeneration assay has been adapted to run on a confocal microscope. Quantitative fluorescence is then used to measure a compounds ability to regenerate neuronal firing.The membrane potential dye di-8-ANEPPS was selected for these experiments. Di-8- ANEPPS is internalized slowly, has a high signal to noise ratio (40:1), has a linear fluorescent response to change in voltage.

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