scholarly journals Confocal Microscopy for Diagnostic Cytology

2004 ◽  
Vol 12 (3) ◽  
pp. 42-43
Author(s):  
Mathilde E. Boon ◽  
Lambrecht P. Kok
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Fluorescent Dyes ◽  
Laser Scanning Confocal Microscopy ◽  
Cervical Cytology ◽  
Scanning Confocal Microscopy ◽  
Additional Information ◽  
Terra Incognita

Laser scanning confocal microscopy has been used to analyze cytological specimens for research purposes. However, for many pathologists, its application in diagnostic cytology still is terra incognita. In our laboratory, confocal microscopy has been used for seven years, mainly to solve problems in cervical cytology, in breast aspirates, and for endometrium cytology.In contrast to histology where we can cut new sections from the block, the cytologic sample is often limited to one smear. Accordingly, when we wish to expand our diagnostic possibilities, we have to extract the desired additional information from that single smear. This is possible by first de-staining the original smear and subsequently re-staining it. In this article, we describe how we restain old slides with the fluorescent dyes Ethidinm Bromide and Eosin, a DNA/protein stain, resulting in staining patterns comparable to the diagnostic Papanicolaou and Giemsa methods in that mere is an optimal distinction between nucleus and cytoplasm.

scholarly journals A Rapid Method for Combined Laser Scanning Confocal Microscopic and Electron Microscopic Visualization of Biocytin or Neurobiotin-labeled Neurons

1998 ◽  
Vol 46 (2) ◽  
pp. 263-273 ◽  
Author(s):  
Xue J. Sun ◽  
Leslie P. Tolbert ◽  
John G. Hildebrand ◽  
Ian A. Meinertzhagen
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Fluorescent Dyes ◽  
Laser Scanning Confocal Microscopy ◽  
Single Step ◽  
Three Dimensions ◽  
Fluorescent Label ◽  
Electron Microscopic ◽  
Scanning Confocal Microscopy ◽  
Long Term Storage

Intracellular recording and dye filling are widely used to correlate the morphology of a neuron with its physiology. With laser scanning confocal microscopy, the complex shapes of labeled neurons in three dimensions can be reconstructed rapidly, but this requires fluorescent dyes. These dyes are neither permanent nor electron dense and therefore do not allow investigation by electron microscopy. Here we report a technique that quickly and easily converts a fluorescent label into a more stable and electron-dense stain. With this technique, a neuron is filled with Neurobiotin or biocytin, reacted with fluorophore-conjugated avidin, and imaged by confocal microscopy. To permit long-term storage or EM study, the fluorescent label is then converted to a stable electron-dense material by a single-step conversion using a commercially available ABC kit. We find that the method, which apparently relies on recognition of avidin's excess biotin binding sites by the biotin–peroxidase conjugate, is both faster and less labor intensive than photo-oxidation procedures in common use. The technique is readily adaptable to immunocytochemistry with biotinylated probes, as we demonstrate using anti-serotonin as an example.

A Multiscale Approach to Multi-Component Wetting

2004 ◽  
Author(s):  
Anne M. Grillet ◽  
Benjamin J. Ash ◽  
Carlton F. Brooks ◽  
John A. Emerson
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Fluorescent Dyes ◽  
Laser Scanning Confocal Microscopy ◽  
Multiscale Approach ◽  
Local Concentration ◽  
Proof Of Concept ◽  
Scanning Confocal Microscopy ◽  
Air Interface ◽  
Spectral Separation

Laser scanning confocal microscopy has been applied to study segregation in multi-component wetting. By labeling the two components of a blend with contrasting fluorescent dyes, the approximate local concentration can be determined from the relative fluorescence intensities. As a proof of concept, a coarsely blended mixture was imaged and parameters were adjusted to achieve good spectral separation of the two components. The technique was then applied to a well-blended drop of the two components and one component was observed to segregate to the air interface.

scholarly journals Application of Laser Scanning Confocal Microscopy for the Visualization of M. tuberculosis in Lung Tissue Samples with Weak Ziehl–Neelsen Staining

2019 ◽  
Vol 8 (8) ◽  
pp. 1185 ◽  
Author(s):  
Maria V. Erokhina ◽  
Larisa N. Lepekha ◽  
Elena E. Voronezhskaya ◽  
Leonid P. Nezlin ◽  
Vadim G. Avdienko ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Laser Scanning ◽  
Fluorescent Antibody ◽  
Laser Scanning Confocal Microscopy ◽  
Emission Analysis ◽  
Tissue Samples ◽  
Specific Fluorescence ◽  
Scanning Confocal Microscopy ◽  
Additional Information ◽  
Size And Morphology

One of the key requirements for the diagnosis of pulmonary tuberculosis is the identification of M. tuberculosis in tissue. In this paper, we present the advantages of specific fluorescent antibody labelling, combined with laser scanning confocal microscopy (LSCM), for the detection of M. tuberculosis in histological specimens of lung tissues. We demonstrate that the application of LSCM allows: (i) The automatic acquisition of images of the whole slice and, hence, the determination of regions for subsequent analysis; (ii) the acquisition of images of thick (20–40 μm) slices at high resolution; (iii) single bacteria identification; and (iv) 3D reconstruction, in order to obtain additional information about the distribution, size, and morphology of solitary M. tuberculosis; as well as their aggregates and colonies, in various regions of tuberculosis inflammation. LSCM allows for the discrimination of the non-specific fluorescence of bacteria-like particles and their aggregates presented in histological lung samples, from the specific fluorescence of labelled M. tuberculosis, using spectrum emission analysis. The applied method was effective in the identification of M. tuberculosis in lung histological samples with weak Ziehl–Neelsen staining. Altogether, combining immunofluorescent labelling with the application of LSCM visualization significantly increases the effectiveness of M. tuberculosis detection.

Laser Scanning Confocal Microscopy and Three-Dimesnsional Reconstruction of the Mitotic Spindles of Unequally Dividing Embryonic Cells

1990 ◽  
Vol 48 (3) ◽  
pp. 166-167
Author(s):  
J. Holy ◽  
G. Schatten
Keyword(s):  
Confocal Microscopy ◽  
Mitotic Spindle ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Confocal Microscopy ◽  
Two Dimensions ◽  
Embryonic Cells ◽  
Mitotic Spindles ◽  
Cleavage Division ◽  
Scanning Confocal Microscopy

One of the classic limitations of light microscopy has been the fact that three dimensional biological events could only be visualized in two dimensions. Recently, this shortcoming has been overcome by combining the technologies of laser scanning confocal microscopy (LSCM) and computer processing of microscopical data by volume rendering methods. We have employed these techniques to examine morphogenetic events characterizing early development of sea urchin embryos. Specifically, the fourth cleavage division was examined because it is at this point that the first morphological signs of cell differentiation appear, manifested in the production of macromeres and micromeres by unequally dividing vegetal blastomeres.The mitotic spindle within vegetal blastomeres undergoing unequal cleavage are highly polarized and develop specialized, flattened asters toward the micromere pole. In order to reconstruct the three-dimensional features of these spindles, both isolated spindles and intact, extracted embryos were fluorescently labeled with antibodies directed against either centrosomes or tubulin.

Observation of Fine Structure in Bicontinuous Phase-Separated Domains of a Polymer Blend by Laser Scanning Confocal Microscopy

2001 ◽  
Vol 34 (15) ◽  
pp. 5186-5191 ◽  
Author(s):  
Hiroshi Jinnai ◽  
Hiroshi Yoshida ◽  
Kohtaro Kimishima ◽  
Yoshinori Funaki ◽  
Yoshitsugu Hirokawa ◽  
...  
Keyword(s):  
Fine Structure ◽  
Confocal Microscopy ◽  
Polymer Blend ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Scanning Confocal Microscopy

scholarly journals The DNA content of trophozoites and cysts of Giardia lamblia by microdensitometric quantitation of Feulgen staining and examination by laser scanning confocal microscopy.

1994 ◽  
Vol 42 (11) ◽  
pp. 1413-1416 ◽  
Author(s):  
S L Erlandsen ◽  
E M Rasch
Keyword(s):  
Confocal Microscopy ◽  
Genome Size ◽  
Dna Content ◽  
Giardia Lamblia ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Evolutionary Development ◽  
Scanning Confocal Microscopy ◽  
Dividing Cells ◽  
C Value

We investigated direct measurement of the DNA content of the parasitic intestinal flagellate Giardia lamblia through quantitation by Feulgen microspectrophotometry and also by visualization of Feulgen-stained DNA chromosomes within dividing cells by laser scanning confocal microscopy. Individual trophozoites of Giardia (binucleate) contained 0.144 +/- 0.018 pg of DNA/cell or 0.072 pg DNA/nucleus. Giardia lamblia cysts (quadranucleate) contained 0.313 +/- 0.003 pg DNA or 0.078 pg DNA/nucleus. The genome size (C) value per nucleus ranged between 6.5-7.1 x 10(7) BP for trophozoites and cysts, respectively. Confocal microscopic examination of Giardia trophozoites undergoing binary fission revealed five chromosome-like bodies within each nucleus. Further information about genome size and DNA content within different Giardia species may help to clarify the pivotal role of these primitive eukaryotic cells in evolutionary development.

The Effect of Chloroquine on the Apoptosis Induced by Cisplatin in Human Gastric Cancer BGC823 Cells

2014 ◽  
Vol 926-930 ◽  
pp. 1124-1127
Author(s):  
Zhen Xun Jin ◽  
Li Li Zhang ◽  
Yan Wang ◽  
Lin Chuan Zeng ◽  
Yang Yu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Confocal Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Combination Group ◽  
Human Gastric Cancer ◽  
Apoptotic Cells ◽  
Toxic Dose ◽  
Scanning Confocal Microscopy ◽  
Mtt Tests

The aim of this study is to investigate the effects and mechanism of chloroquine (CQ) on the apoptosis induced by cisplatin in human gastric cancer BGC823 cells. MTT assay was used to detect the state of cell growth. The appearances of cellular apoptosis were detected by laser scanning confocal microscopy and light microscopy. The expressions of LC3 and p62 were detected by laser scanning confocal microscopy. MTT tests showed that the non-toxic dose of CQ could increase the inhibition rate of BGC823 cells induced by cisplatin. Under the light microscope, the ratio of apoptotic cells in the group treated with non-toxic dose of CQ combined with cisplatin was higher than that in the group treated with cisplatin alone. Hoechst33342 staining showed that the ratio of apoptotic cells in the combination group was higher than that in the cisplatin group. The expression and colocalization of LC3 and p62 proteins were significantly increased in the combination group. These results indicate that CQ can enhance the cell apoptosis induced by cisplatin in BGC823 cells, which is through the inhibition of autophagy.

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