A method to compensate for light attenuation with depth in three-dimensional DNA image cytometry using a confocal scanning laser microscope

1995 ◽  
Vol 177 (2) ◽  
pp. 108-114 ◽  
Author(s):  
A. LILJEBORG ◽  
M. CZADER ◽  
A. PORWIT
Keyword(s):  
Light Attenuation ◽  
Three Dimensional ◽  
Image Cytometry ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscope ◽  
Confocal Scanning ◽  
Confocal Scanning Laser ◽  
Dna Image Cytometry

scholarly journals Three-dimensional DNA image cytometry by confocal scanning laser microscopy in thick tissue blocks

Cytometry ◽  
1991 ◽  
Vol 12 (6) ◽  
pp. 511-524 ◽  
Author(s):  
Jean Paul Rigaut ◽  
Jany Vassy ◽  
Paulette Herlin ◽  
Françgoise Duigou ◽  
Eric Masson ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Image Cytometry ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscopy ◽  
Laser Microscopy ◽  
Scanning Laser Microscopy ◽  
Confocal Scanning ◽  
Confocal Scanning Laser ◽  
Dna Image Cytometry

Three-dimensional images of DAB-labeled neurons by confocal scanning laser microscopy

1989 ◽  
Vol 47 ◽  
pp. 144-145
Author(s):  
JS Deitch ◽  
KL Smith ◽  
C Lee ◽  
JW Swann ◽  
JN Turner
Keyword(s):  
Three Dimensional ◽  
Neuronal Morphology ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscopy ◽  
Confocal Scanning Laser Microscope ◽  
Basal Dendrites ◽  
Confocal Scanning ◽  
Three Dimensional Images ◽  
Confocal Scanning Laser ◽  
Areas Of Interest

The ability to correlate neuronal morphology and physiology has been greatly advanced by intracellular labeling through the recording pipette. However, visualizing the filled neuron required physically sectioning and reconstructing areas of interest, often resulting in figures that are two-dimensional. We have visualized the three-dimensional morphology of filled neurons reacted with nickel-intensified diaminobenzidine (DAB/Ni) using the confocal scanning laser microscope (CSLM).Neurons in slices of rat hippocampus were filled with biocytin, fixed in 4% paraformaldehyde, incubated in avidin-HRP (1:200) in 0.5% Triton X-100, and reacted with DAB in 0.04% nickel ammonium sulfate. Optical sections and three-dimensional images were recorded by using a Bio Rad MRC-500 CSLM with an argon ion laser.Biocytin filled all aspects of the neuron, including fine axons and spines. Fig. 1 is a conventional micrograph of a single neuron labeled with DAB/Ni. Figs. 2a and b are stereo pairs of the apical and basal dendrites of the neuron in Fig. 1.

Measurements in 3D images

1991 ◽  
Vol 49 ◽  
pp. 408-409
Author(s):  
John C. Russ
Keyword(s):  
Three Dimensional ◽  
Image Plane ◽  
X Rays ◽  
Traditional Use ◽  
3D Images ◽  
Confocal Scanning Laser Microscope ◽  
Hard Materials ◽  
Depth Direction ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

Three-dimensional (3D) images consisting of arrays of voxels can now be routinely obtained from several different types of microscopes. These include both the transmission and emission modes of the confocal scanning laser microscope (but not its most common reflection mode), the secondary ion mass spectrometer, and computed tomography using electrons, X-rays or other signals. Compared to the traditional use of serial sectioning (which includes sequential polishing of hard materials), these newer techniques eliminate difficulties of alignment of slices, and maintain uniform resolution in the depth direction. However, the resolution in the z-direction may be different from that within each image plane, which makes the voxels non-cubic and creates some difficulties for subsequent analysis.

Fading of the diaminobenzidine reaction product in the confocal scanning laser microscope

1989 ◽  
Vol 47 ◽  
pp. 146-147
Author(s):  
JS Deitch ◽  
KL Smith ◽  
JW Swann ◽  
JN Turner
Keyword(s):  
Pyramidal Neurons ◽  
Light And Electron Microscopy ◽  
Scanning Laser ◽  
Laser Exposure ◽  
Confocal Scanning Laser Microscope ◽  
Fluorescent Markers ◽  
Before And After ◽  
Staining Protocol ◽  
Confocal Scanning ◽  
Confocal Scanning Laser

Neurons labeled with horseradish peroxidase and reacted with diaminobenzidine (DAB) can be imaged using a confocal scanning laser microscope (CSLM) in the reflection mode. In contrast to fluorescent markers, the DAB reaction product is thought to be stable and can be observed by both light and electron microscopy. We have investigated the sensitivity of the DAB reaction product to laser irradiation, and present the spectrophotometric properties of DAB before and after exposure in the CSLM.Pyramidal neurons in slices of rat hippocampus were injected with biocytin (a biotin-lysine complex), fixed overnight in 4% paraformaldehyde, and vibratome sectioned at 75 μm. Biocytin was detected with avidin-HRP (1:200) in 0.5% Triton X-100, incubated in DAB (0.5 mg/ml) with or without 0.04% nickel ammonium sulfate (Ni), dehydrated, and imaged in a Bio Rad MRC-500 CSLM with an argon ion laser (488 and 514 nm). Spectrophotometric measurements of the soma were made on a Zeiss microspectrophotometer, as a function of laser exposure (100-1000 scans) and staining protocol.

Diagnostic Revolution of Microhematuria by Real Time Confocal Scanning Laser Microscope: Hyodo-Iino-Miyagawa Method, Third Report

Nephron ◽  
1995 ◽  
Vol 70 (2) ◽  
pp. 171-179 ◽  
Author(s):  
Toru Hyodo ◽  
Ikuo Miyagawa ◽  
Akihiro Iino ◽  
Koji Ono ◽  
Tsutomu Kuomi ◽  
...  
Keyword(s):  
Real Time ◽  
Scanning Laser ◽  
Confocal Scanning Laser Microscope ◽  
Confocal Scanning ◽  
Confocal Scanning Laser
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