Digital imaging microscopy: the marriage of spectroscopy and the solid state CCD camera

1991 ◽  
Author(s):  
Thomas M. Jovin ◽  
Donna J. Arndt-Jovin
Keyword(s):  
Solid State ◽  
Digital Imaging ◽  
Ccd Camera ◽  
Digital Imaging Microscopy

Multi-Mode Light Microscopy of Microtubule and Endoplasmic Reticulum Dynamics in Migrating Newt Epithelial Cells.

1997 ◽  
Vol 3 (S2) ◽  
pp. 211-212
Author(s):  
C. M. Waterman-Storer ◽  
E. D. Salmon
Keyword(s):  
Digital Imaging ◽  
Imaging System ◽  
Ccd Camera ◽  
Diaphragm Position ◽  
Multiple Band ◽  
Fluorescence Light ◽  
Band Pass ◽  
Cooled Ccd ◽  
System 1 ◽  
Multi Mode

We have developed a multi-mode digital imaging system (1-3) which acquires images with a 12 bit cooled CCD camera. A multiple band pass dichromatic mirror and robotically controlled excitation filter wheels provide rapid wavelength selection for epi-fluorescence with DAPI, fluorescein or GFP and X-rhodamine fluorophores while maintaining image registration on the cooled CCD detector. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. A robotically controlled emission filter wheel in front of the CCD camera inserts an analyzer in the light path for DIC imaging. To maximize fluorescence light intensity, the analyzer is removed and an optical flat of equivalent optical thickness is inserted for fluorescence imaging. A slider is inserted at the field diaphragm position of the fluorescence epi-illuminator to provide in-focus slit and spot targets for 360 nm photoactivation of “caged” fluorophores. The microscope system is robotically controlled and image acquisition and analysis is performed using MetaMorph™ digital imaging software.

Fluorescence digital imaging microscopy in cell biology

Science ◽  
1985 ◽  
Vol 230 (4723) ◽  
pp. 247-256 ◽  
Author(s):  
D. Arndt-Jovin ◽  
M Robert-Nicoud ◽  
S. Kaufman ◽  
T. Jovin
Keyword(s):  
Digital Imaging ◽  
Cell Biology ◽  
Digital Imaging Microscopy

scholarly journals Fluorescence ratio measurements of double-labeled probes for multiple in situ hybridization by digital imaging microscopy

Cytometry ◽  
1992 ◽  
Vol 13 (8) ◽  
pp. 839-845 ◽  
Author(s):  
P. M. Nederlof ◽  
S. van der Flier ◽  
J. Vrolijk ◽  
H. J. Tanke ◽  
A. K. Raap
Keyword(s):  
In Situ Hybridization ◽  
Digital Imaging ◽  
Fluorescence Ratio ◽  
Digital Imaging Microscopy

scholarly journals Microtubule dynamics and chromosome motion visualized in living anaphase cells.

1988 ◽  
Vol 106 (4) ◽  
pp. 1185-1192 ◽  
Author(s):  
G J Gorbsky ◽  
P J Sammak ◽  
G G Borisy
Keyword(s):  
Chromosome Segregation ◽  
Digital Imaging ◽  
Living Cells ◽  
Microtubule Dynamics ◽  
Chromosome Movement ◽  
Animal Cells ◽  
Digital Imaging Microscopy ◽  
Opposite Pole ◽  
Anaphase B ◽  
Chromosome Motion

Chromosome segregation in most animal cells is brought about through two events: the movement of the chromosomes to the poles (anaphase A) and the movement of the poles away from each other (anaphase B). Essential to an understanding of the mechanism of mitosis is information on the relative movements of components of the spindle and identification of sites of subunit loss from shortening microtubules. Through use of tubulin derivatized with X-rhodamine, photobleaching, and digital imaging microscopy of living cells, we directly determined the relative movements of poles, chromosomes, and a marked domain on kinetochore fibers during anaphase. During chromosome movement and pole-pole separation, the marked domain did not move significantly with respect to the near pole. Therefore, the kinetochore microtubules were shortened by the loss of subunits at the kinetochore, although a small amount of subunit loss elsewhere was not excluded. In anaphase A, chromosomes moved on kinetochore microtubules that remained stationary with respect to the near pole. In anaphase B, the kinetochore fiber microtubules accompanied the near pole in its movement away from the opposite pole. These results eliminate models of anaphase in which microtubules are thought to be traction elements that are drawn to and depolymerized at the pole. Our results are compatible with models of anaphase in which the kinetochore fiber microtubules remain anchored at the pole and in which microtubule dynamics are centered at the kinetochore.

Characterization of Topotecan-Mediated Redistribution of DNA Topoisomerase I by Digital Imaging Microscopy

1998 ◽  
Vol 241 (2) ◽  
pp. 332-339 ◽  
Author(s):  
Randy M. Wadkins ◽  
Mary K. Danks ◽  
Lisa Horowitz ◽  
Sharyn D. Baker
Keyword(s):  
Digital Imaging ◽  
Topoisomerase I ◽  
Dna Topoisomerase I ◽  
Dna Topoisomerase ◽  
Digital Imaging Microscopy

Altered reactivity of hemolysate-treated cultured smooth-muscle cells from rabbit basilar artery determined by digital imaging microscopy

1991 ◽  
Vol 75 (1) ◽  
pp. 82-90 ◽  
Author(s):  
Yoshihiro Takanashi ◽  
Kazuhiko Fujitsu ◽  
Satoshi Fujii ◽  
Takeo Kuwabara
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Digital Imaging ◽  
Muscle Cells ◽  
Blocking Agent ◽  
Interference Microscopy ◽  
Morphological Alteration ◽  
Ultrastructural Changes ◽  
Digital Imaging Microscopy ◽  
Basilar Arteries

✓ During culture, smooth-muscle cells obtained from rabbit basilar arteries were examined for contractile activity by means of differential interference microscopy with a video analysis system (digital imaging microscopy system). This system proved useful for observing the contraction and ultrastructural changes of the living cells. Hemolysate-treated cells showed augmented responses to 5-hydroxytryptamine and leukotriene C4, but not to KCl. This augmented response diminished gradually during the culture period. Both a phospholipase C blocking agent, 2-nitro-4-carboxyphenyl-n,n-diphenylcarbamate (NCDC), and a myosin light chain kinase blocking agent, 1-(5-chloronaphthalenesulfonyl)-1 H-hexahydro-1,4-diazepine (ML-9), suppressed this augmented response. Protein kinase C activity of the cells, as measured by Western blot analysis, did not increase during the period of culture with hemolysate. The results obtained suggest that hemolysate had the following effects on the cells: 1) acute but gradual contraction of the cells; 2) augmentation of cellular responses to vasoactive agents; and 3) progressive contraction and morphological alteration of the cells. Possible mechanisms by which hemolysate exerts these effects are discussed, taking into consideration the interrelationship between these effects.

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