scholarly journals Giemsa as a fluorescent stain for mineralized bone.

1994 ◽  
Vol 42 (5) ◽  
pp. 677-680 ◽  
Author(s):  
J N Bradbeer ◽  
M Riminucci ◽  
P Bianco
Keyword(s):  
Fluorescence Microscopy ◽  
Differential Staining ◽  
Elastic Fibers ◽  
Eosin Y ◽  
Fluorescent Staining ◽  
Tissue Sections ◽  
Confocal Fluorescence ◽  
Differential Binding ◽  
Fluorescent Stain ◽  
New Applications

We present evidence for a previously unrecognized differential staining effect of Giemsa solution in fluorescence microscopy. The effect consists of selective fluorescent staining of mineralized bone (and elastic fibers) in tissue sections and, like the classical Romanowsky effect, is based on the differential binding of Eosin Y to tissue structures in the presence of Azur II and Methylene Blue. This effect opens the way to new applications of the Giemsa solution in fluorescence microscopy and in confocal fluorescence microscopy.

scholarly journals Ex Vivo Confocal Fluorescence Microscopy for Rapid Evaluation of Tissues in Surgical Pathology Practice

2017 ◽  
Vol 142 (3) ◽  
pp. 396-401 ◽  
Author(s):  
Savitri Krishnamurthy ◽  
Andrea Cortes ◽  
Mirtha Lopez ◽  
Michael Wallace ◽  
Sharjeel Sabir ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Acridine Orange ◽  
Ex Vivo ◽  
Imaging Techniques ◽  
Surgical Pathology ◽  
Confocal Fluorescence Microscopy ◽  
Tissue Preparation ◽  
Tissue Sections ◽  
Confocal Fluorescence ◽  
Hematoxylin Eosin

Context.— Optical imaging techniques are currently available for imaging tissues without the need for any type of extensive tissue preparation. There are several applications for their potential use in surgical pathology practice. Objective.— To evaluate the feasibility of using a confocal fluorescence microscopy (CFM) platform for ex vivo examination of tissues obtained from surgical resections of breast, lung, kidney, and liver. Design.— Tissue fragments (0.5–1.0 cm) were immersed in 0.6 mM acridine orange for 6 seconds and imaged using a CFM platform at a 488-nm wavelength. The imaged tissues were subsequently fixed in formalin and processed routinely to generate hematoxylin-eosin–stained tissue sections. Mosaics of the grayscale CFM images were studied at different magnifications for recognition of the tissue and were compared with conventional histopathologic examination of hematoxylin-eosin tissue sections. Results.— We imaged 55 tissue fragments obtained from 16 breast (29%), 18 lung (33%), 14 kidney (25%), and 7 liver (13%) surgical excision specimens. Acridine orange labeled the nuclei, creating the contrast between nucleus and cytoplasm and thereby recapitulating the tissue architecture. We could obtain CFM images of good quality within 5 to 10 minutes that allowed recognition of the cytomorphologic details for categorization of the imaged tissue and were similar to histologic examination of hematoxylin-eosin tissue sections. Conclusions.— The ease and speed of acquisition of CFM images together with the resolution and resemblance of the CFM images to hematoxylin-eosin sections suggest that the CFM platform has excellent potential for use in surgical pathology practice.

scholarly journals Confocal Fluorescence Microscopy Platform Suitable for Rapid Evaluation of Small Fragments of Tissue in Surgical Pathology Practice

2018 ◽  
Vol 143 (3) ◽  
pp. 305-313 ◽  
Author(s):  
Savitri Krishnamurthy ◽  
Kechen Ban ◽  
Kenna Shaw ◽  
Gordon Mills ◽  
Rahul Sheth ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Predictive Value ◽  
Surgical Excision ◽  
Surgical Pathology ◽  
Confocal Fluorescence Microscopy ◽  
Rapid Evaluation ◽  
Tissue Sections ◽  
Confocal Fluorescence ◽  
Hematoxylin Eosin

Context.— Rapid advances in the fields of biophotonics, computer science, and instrumentation have allowed for high-resolution imaging of biologic tissues. Objective.— To evaluate the quality of images from an optimized confocal fluorescence microscopy (CFM) platform for rapid evaluation of small fragments of tissue, compared with hematoxylin-eosin staining. Design.— Tissue fragments (up to 1.0 × 0.3 cm) were stained with 0.6 mM acridine orange for 60 seconds and imaged using a CFM platform at 488-nm and 785-nm wavelength. The imaged tissues were then fixed in formalin and processed to generate hematoxylin-eosin–stained tissue sections. The quality of CFM images was scored on a scale of 0 to 3 on the basis of the percentage of the CFM images with recognizable tissue architecture (0, 0%; 1, <20%; 2, 20%–50%; 3, >50%). The diagnoses made using CFM images were compared with those made using histopathologic analysis of the hematoxylin-eosin–stained tissue sections. Results.— We imaged 118 tissue fragments obtained from 40 breast, 23 lung, 39 kidney, and 16 liver surgical excision specimens. We acquired CFM images in 2 to 3 minutes; 95.8% (113 of 118) of images showed a quality score of 3, and 4.2% (5 of 118) had a score of 2. We achieved a sensitivity of 95.5%, specificity of 97.3%, positive predictive value of 95.5%, and negative predictive value of 97.3%. Conclusions.— Our results demonstrate the suitability of the CFM platform for rapid and accurate evaluation of small tissue fragments in surgical pathology practice.

scholarly journals Analysis of Polyhydroxyalkanoates Granules in Haloferax mediterranei by Double-Fluorescence Staining with Nile Red and SYBR Green by Confocal Fluorescence Microscopy

Polymers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1582
Author(s):  
Verónica Cánovas ◽  
Salvador Garcia-Chumillas ◽  
Fuensanta Monzó ◽  
Lorena Simó-Cabrera ◽  
Carmen Fernández-Ayuso ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Culture Media ◽  
Nile Red ◽  
Sybr Green ◽  
Confocal Fluorescence Microscopy ◽  
Fluorescence Staining ◽  
Haloferax Mediterranei ◽  
Optimized Protocol ◽  
Confocal Fluorescence ◽  
Pha Production

Haloferaxmediterranei is a haloarchaeon of high interest in biotechnology because it produces and mobilizes intracellular polyhydroxyalkanoate (PHA) granules during growth under stress conditions (limitation of phosphorous in the culture media), among other interesting metabolites (enzymes, carotenoids, etc.). The capability of PHA production by microbes can be monitored with the use of staining-based methods. However, the staining of haloarchaea cells is a challenging task; firstly, due to the high ionic strength of the medium, which is inappropriate for most of dyes, and secondly, due to the low permeability of the haloarchaea S-layer to macromolecules. In this work, Haloferax mediterranei is used as a halophilic archaeon model to describe an optimized protocol for the visualization and analysis of intracellular PHA granules in living cells. The method is based on double-fluorescence staining using Nile red and SYBR Green by confocal fluorescence microscopy. Thanks to this method, the capability of PHA production by new haloarchaea isolates could be easily monitored.

Confocal fluorescence microscopy with the tandem scanning light microscope

1989 ◽  
Vol 94 (4) ◽  
pp. 617-624
Author(s):  
S.J. Wright ◽  
J.S. Walker ◽  
H. Schatten ◽  
C. Simerly ◽  
J.J. McCarthy ◽  
...  
Keyword(s):  
High Resolution ◽  
Fluorescence Microscopy ◽  
Light Microscope ◽  
Three Dimensional ◽  
Three Dimensions ◽  
Complex Structures ◽  
Charge Coupled Device ◽  
Confocal Fluorescence ◽  
Cooled Ccd ◽  
Cellular Components

Applications of the tandem scanning confocal microscope (TSM) to fluorescence microscopy and its ability to resolve fluorescent biological structures are described. The TSM, in conjunction with a cooled charge-coupled device (cooled CCD) and conventional epifluorescence light source and filter sets, provided high-resolution, confocal data, so that different fluorescent cellular components were distinguished in three dimensions within the same cell. One of the unique features of the TSM is the ability to image fluorochromes excited by ultraviolet light (e.g. Hoechst, DAPI) in addition to fluorescein and rhodamine. Since the illumination is dim, photobleaching is insignificant and prolonged viewing of living specimens is possible. Series of optical sections taken in the Z-axis with the TSM were reproduced as stereo images and three-dimensional reconstructions. These data show that the TSM is potentially a powerful tool in fluorescence microscopy for determining three-dimensional relationships of complex structures within cells labeled with multiple fluorochromes.

Novel nontoxic mitochondrial probe for confocal fluorescence microscopy

2006 ◽  
Vol 11 (3) ◽  
pp. 034014 ◽  
Author(s):  
Silvia Versari ◽  
Anna Maria Villa ◽  
Alessandro Villa ◽  
Silvia Maria Doglia ◽  
Giorgio A. Pagani ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Confocal Fluorescence Microscopy ◽  
Confocal Fluorescence
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