scholarly journals A novel technique for quantitative autoradiography of labeled histological specimens.

1981 ◽  
Vol 29 (1) ◽  
pp. 79-83 ◽  
Author(s):  
L M Partlow ◽  
L G Bush ◽  
L J Stensaas ◽  
R P Kesner
Keyword(s):  
Arbitrary Shape ◽  
Quantitative Method ◽  
Quantitative Autoradiography ◽  
Histological Structure ◽  
Image Analyzer ◽  
Internal Standards ◽  
Tissue Sections ◽  
Novel Technique ◽  
Entire Specimen ◽  
Tissue Specimens

A novel quantitative method is described that makes it possible to routinely and accurately quantify contact autoradiographs of tissue sections in terms of actual amounts of radioactivity per square millimeter. Such determinations can either be made on the entire specimen or on any selected area of arbitrary shape that might correspond to a particular histological structure. This method utilizes the tissue specimens themselves as internal standards and requires the availability of a computerized image analyzer.

scholarly journals Discrimination of Aspergillosis, Mucormycosis, Fusariosis, and Scedosporiosis in Formalin-Fixed Paraffin-Embedded Tissue Specimens by Use of Multiple Real-Time Quantitative PCR Assays

2016 ◽  
Vol 54 (11) ◽  
pp. 2798-2803 ◽  
Author(s):  
Elham Salehi ◽  
Mohammad T. Hedayati ◽  
Jan Zoll ◽  
Haleh Rafati ◽  
Maryam Ghasemi ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Aspergillus Flavus ◽  
Ffpe Tissue ◽  
Content Type ◽  
Real Time Quantitative Pcr ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Formalin Fixed

In a retrospective multicenter study, 102 formalin-fixed paraffin-embedded (FFPE) tissue specimens with histopathology results were tested. Two 4- to 5-μm FFPE tissue sections from each specimen were digested with proteinase K, followed by automated nucleic acid extraction. Multiple real-time quantitative PCR (qPCR) assays targeting the internal transcribed spacer 2 (ITS2) region of ribosomal DNA, using fluorescently labeled primers, was performed to identify clinically important genera and species of Aspergillus , Fusarium , Scedosporium , and the Mucormycetes . The molecular identification was correlated with results from histological examination. One of the main findings of our study was the high sensitivity of the automated DNA extraction method, which was estimated to be 94%. The qPCR procedure that was evaluated identified a range of fungal genera/species, including Aspergillus fumigatus , Aspergillus flavus , Aspergillus terreus , Aspergillus niger , Fusarium oxysporum , Fusarium solani , Scedosporium apiospermum , Rhizopus oryzae , Rhizopus microsporus , Mucor spp., and Syncephalastrum . Fusarium oxysporum and F. solani DNA was amplified from five specimens from patients initially diagnosed by histopathology as having aspergillosis. Aspergillus flavus , S. apiospermum , and Syncephalastrum were detected from histopathological mucormycosis samples. In addition, examination of four samples from patients suspected of having concomitant aspergillosis and mucormycosis infections resulted in the identification of two A. flavus isolates, one Mucor isolate, and only one sample having both R. oryzae and A. flavus . Our results indicate that histopathological features of molds may be easily confused in tissue sections. The qPCR assay used in this study is a reliable tool for the rapid and accurate identification of fungal pathogens to the genus and species levels directly from FFPE tissues.

Automated Method for the Analysis of Riboflavin in Milk, with Application to Other Selected Foods

1974 ◽  
Vol 57 (5) ◽  
pp. 1085-1088
Author(s):  
James R Kirk
Keyword(s):  
Standard Deviation ◽  
Continuous Flow ◽  
Quantitative Method ◽  
Green Fluorescence ◽  
Internal Standards ◽  
Automated Method ◽  
Sodium Hydrosulfite ◽  
Automated Technique ◽  
Good Agreement

Abstract A continuous flow automated technique was developed for the determination of riboflavin in milk. The determination is based on the measurement of the natural yellow-green fluorescence of riboflavin at an excitation of 436 nm and emission of 510 nm. Blank values are determined for each sample after sodium hydrosulfite reduction of the riboflavin. Mean recovery and standard deviation for riboflavin in milk determined by the continuous flow procedure using internal standards were 9 7% and ± 2.42%, respectively. The recovery value was in good agreement with that determined using a manual procedure, while the standard deviation was 33% less than that found when using the manual procedure. The results from this study indicate that the continuous flow automated procedure for the determination of riboflavin in milk is a simple, quantitative method which eliminates many of the time-consuming analytical steps.

A Novel Technique for the Detection of LncRNAs on Tissue Sections

2020 ◽  
pp. 237-243
Author(s):  
Andrew E. Massey ◽  
Manish K. Tripathi ◽  
Murali M. Yallapu ◽  
Meena Jaggi ◽  
Subhash C. Chauhan
Keyword(s):  
Tissue Sections ◽  
Novel Technique

FEATURES OF THE PATHOMORPHOLOGICAL DIAGNOSIS OF MICROCRYSTALLINE ARTHROPATHIES IN THE PRACTICE OF SURGICAL MATERIAL EXAMINATION

2020 ◽  
Vol 58 (3) ◽  
pp. 286-289
Author(s):  
N. S. Migalkin ◽  
T. A. Stupina ◽  
A. V. Kaminsky ◽  
D. S. Mokhovikov ◽  
D. A. Shabalin ◽  
...  
Keyword(s):  
Staining Technique ◽  
Calcium Pyrophosphate ◽  
Alcohol Solution ◽  
Formalin Fixation ◽  
Histological Techniques ◽  
Tissue Sections ◽  
Stereo Microscope ◽  
Hematoxylin And Eosin ◽  
Tissue Specimens ◽  
Msu Crystals

The development of microcrystalline arthritides is most frequently associated with the formation of monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals. Their identification is of crucial importance in recognizing these diseases. Objective: to determine the possibilities of histological techniques in identifying MSU and CPP crystals and to evaluate the effectiveness of the techniques. Subjects and methods. Twenty-four tissue blocks (fragments of the affected areas of the elbow joint, the interphalangeal joint of the index finger, and hip joint) from 7 patients were examined. Paraffin sections were stained with a 0.5% alcohol solution of eosin, as well as with hematoxylin and eosin. Tissue specimens were examined and digitized using an AxioScope.A1 stereo microscope with Zenblue software (Carl Zeiss MicroImaging GmbH, Germany). Results and discussion. When staining the tissue sections with hematoxylin and eosin, microcrystals were not visualized; the major portions of MSU crystals was dissolved during fixation and staining, whereas CPP crystals were masked with hematoxylin as focal basophilic aggregates. The staining technique with an alcohol solution of eosin and short formalin fixation (within 12 hours) made it possible to avoid dissolution of MSU crystals and to visualize both MSU and CPP crystals, and to determine their shape and color. Conclusion. Light microscopy of the tissue sections stained with an alcohol solution of eosin along with short formalin fixation is a reliable method to differentiate MSU and CPP crystals. In patients undergoing endoprosthetic replacement, the significance of this technique for the pathomorphological study of surgical material consists in assessing inflammatory activity and in eliminating a disease, such as microcrystalline arthropathy.

scholarly journals Laser Capture Microdissection in Dentistry

2010 ◽  
Vol 2010 ◽  
pp. 1-8 ◽  
Author(s):  
Uraiwan Chokechanachaisakul ◽  
Tomoatsu Kaneko ◽  
Takashi Okiji ◽  
Reika Kaneko ◽  
Hideaki Suda ◽  
...  
Keyword(s):  
Laser Capture Microdissection ◽  
Gene Expression Analysis ◽  
Cell Types ◽  
Specific Cell ◽  
Tissue Sections ◽  
Rna Analysis ◽  
Molecular Biological Analysis ◽  
Polymerase Chain ◽  
Tissue Specimens ◽  
Laser Capture

Laser capture microdissection (LCM) allows for the microscopic procurement of specific cell types from tissue sections that can then be used for gene expression analysis. According to the recent development of the LCM technologies and methodologies, the LCM has been used in various kinds of tissue specimens in dental research. For example, the real-time polymerase-chain reaction (PCR) can be performed from the formaldehyde-fixed, paraffin-embedded, and immunostained sections. Thus, the advance of immuno-LCM method allows us to improve the validity of molecular biological analysis and to get more accurate diagnosis in pathological field in contrast to conventional LCM. This paper is focused on the presentation and discussion of the existing literature that covers the fields of RNA analysis following LCM in dentistry.

scholarly journals Interphase cytogenetics using fluorescence in situ hybridization: an overview of its application to diffuse and solid tissue

1997 ◽  
Vol 20 (1) ◽  
pp. 97-106 ◽  
Author(s):  
Christine Hackel ◽  
Marileila Varella-Garcia
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Bone Marrow Cells ◽  
Isolated Cells ◽  
Interphase Cytogenetics ◽  
Tissue Samples ◽  
Solid Tissue ◽  
Tissue Sections ◽  
Tissue Specimens

Interphase cytogenetics, utilizing fluorescence in situ hybridization (FISH) techniques, has been successfully applied to diffuse and solid tissue specimens. Most studies have been performed on isolated cells, such as blood or bone marrow cells; a few have been performed on cells from body fluids, such as amniotic fluid, urine, sperm, and sputum. Mechanically or chemically disaggregated cells from solid tissues have also been used as single cell suspensions for FISH. Additionally, intact organized tissue samples represented by touch preparations or thin tissue sections have been used, especially in cancer studies. Advantages and pitfalls of application of FISH methodology to each type of specimen and some significant biological findings achieved are illustrated in this overview.

scholarly journals Roles of Proliferation and Angiogenesis in Locally Aggressive Biologic Behavior of Ameloblastoma versus Ameloblastic Fibroma

2022 ◽  
Author(s):  
Amr Ibrahim ◽  
Emad Elqalshy ◽  
Ahmed El-Mohamadi ◽  
Kamal Abd El-Rahman ◽  
Magdy Alazzazi
Keyword(s):  
Protein Expression ◽  
Wide Distribution ◽  
Vascular Density ◽  
Image Analyzer ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Ameloblastic Fibroma ◽  
Formalin Fixed Paraffin Embedded ◽  
Embedded Blocks ◽  
The Mean

Background: The present study was carried out to evaluate the roles of proliferation and angiogenesis in locally aggressive biologic behavior of ameloblastoma versus ameloblastic fibroma; Methods: 30 formalin-fixed paraffin embedded blocks (15 cases of ameloblastoma & 15 cases of ameloblastic fibroma) were used. To evaluate the proliferation, the tissue sections were stained with AgNORs stain. CD105 was used as immunohistochemical marker of angiogenesis. Quantitative evaluations of AgNORs were performed. The mean vascular density was evaluated as a measure for CD105 protein expression by using image analyzer computer system; Results: The mean number of AgNORs dots per nucleus was significantly higher in ameloblastoma as compared to ameloblastic fibroma. Also, the protein level of CD105 showed positive expression and wide distribution that the mean vascular density was significantly higher in ameloblastoma as compared to ameloblastic fibroma; Conclusion: Quantitative evaluation of AgNORs stain & the mean vascular density utilizing CD105 protein expression may reflect a higher proliferative activity and a more locally aggressive biologic behavior of ameloblastoma when compared to ameloblastic fibroma, that other factors may be involved in biologic behavior of ameloblastic fibroma.

scholarly journals Illuminating chromatin compaction in live cells and fixed tissues using SiR-DNA fluorescence lifetime

2020 ◽  
Author(s):  
Colin Hockings ◽  
Chetan Poudel ◽  
Kevin A. Feeney ◽  
Clara L. Novo ◽  
Mehdi S. Hamouda ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Quantitative Method ◽  
Embryonic Stem ◽  
Nuclear Architecture ◽  
Live Cells ◽  
Fixed Tissue ◽  
Chromatin Compaction ◽  
Tissue Sections ◽  
Cell Transition ◽  
Live Cancer Cell

The global compaction state of chromatin in a nucleus is an important component of cell identity that has been difficult to measure. We have developed a quantitative method to measure the chromatin compaction state in both live and fixed cells, without the need for genetic modification, using the fluorescence lifetime of SiR-DNA dye. After optimising this method using live cancer cell lines treated to induce chromatin compaction or decompaction, we observed chromatin compaction in differentiating epithelial cells in fixed tissue sections, as well as local decompaction foci that may represent transcription factories. In addition, we shed new light on chromatin decompaction during embryonic stem cell transition out of their naïve pluripotent state. This method will be useful to studies of nuclear architecture, and may be easy, cheap, and accessible enough to serve as a general assay of ‘stem-ness’.

scholarly journals Plasmacytoma in patients with multiple myeloma: morphology and immunohistochemistry

2019 ◽  
Author(s):  
Maiia Firsova ◽  
Larisa Mendeleeva ◽  
Savchenko Valeri ◽  
Alla Kovrigina ◽  
Maxim Solovev
Keyword(s):  
Plasma Cells ◽  
Bone Destruction ◽  
Extramedullary Plasmacytoma ◽  
Histological Structure ◽  
Ki 67 ◽  
Mean Values ◽  
Significant Difference ◽  
High Proliferative Activity ◽  
The Mean ◽  
Tissue Specimens

Abstract Background. To study the histological structure and immunohistochemical (IHC) parameters of the plasmacytoma tumor substrate in patients with MM. Methods. The study included 21 patients (10 men/11 women) aged 23 to 73 years old with a newly diagnosed MM complicated by plasmacytoma. Bone plasmacytoma was diagnosed in 14 patients, and extramedullary plasmacytoma was diagnosed in 7 patients. Plasmacytoma tissue specimens were examined using a LEICA DM4000B microscope. Anti-CD56, CD166, CXCR4, Ki-67, c-MYC antibody panels were used for the IHC study of plasmacytoma biopsy. Results. When comparing the morphology of bone and extramedullary plasmacytoma, no significant differences were revealed, however, when the substrate of the extramedullary plasmacytoma was compared to the bone plasmacytoma substrate it was more often represented by tumor cells with immature morphology (57.1% vs. 28.6%). We founded a significant difference in the expression of CD166 in bone and extramedullary plasmacytoma. It was shown that the mean values ​​of the CD166 expression in bone plasmacytoma cells were significantly higher (p = 0.033) and amounted to 36.29 ± 7.61% versus 9.57 ± 8.46% in extramedullary plasmacytoma cells. We noticed that in the cells of the extramedullary plasmacytoma, there were higher values ​​in the Ki-67 index than observed in comparison to the cells of the bone plasmacytoma independent of the cells’morphology. Conclusion. The mechanisms involved in the dissemination of tumor plasma cells are currently unexplored. Our study revealed significant differences in the IHC parameters of the tumor substrate from extramedullary and bone plasmacytoma. Thus, the expression of the CD166 in extramedullary plasmacytoma cells is almost 4 times lower than that in bone plasmacytoma cells, which may indicate the involvement of CD166 in the mechanisms of bone destruction. A high proliferative activity of extramedullary plasmacytoma cells was shown when compared to bone plasmacytoma cells.

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