scholarly journals Optimization of processing female genital tissue samples for lymphocyte analysis by flow cytometry

2020 ◽  
Vol 83 (5) ◽  
Author(s):  
Kevin A. Stoner ◽  
May A. Beamer ◽  
Hilary A. Avolia ◽  
Leslie A. Meyn ◽  
Sharon L. Hillier ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tissue Samples ◽  
Genital Tissue ◽  
Female Genital

Flow Cytometrically Determined DNA Content of Breast Carcinoma and Benign Lesions: Correlations with Histopathological Parameters

1986 ◽  
Vol 72 (2) ◽  
pp. 171-177 ◽  
Author(s):  
Raffaella Uccelli ◽  
Alberto Calugi ◽  
Donato Forte ◽  
Francesco Mauro ◽  
Paolo Polonio-Balbi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Cell Subpopulation ◽  
Aneuploid Cell ◽  
Breast Carcinomas ◽  
Benign Lesions ◽  
Dna Index ◽  
Tissue Samples ◽  
Normal Tissues ◽  
Relative Dna Content

The relative DNA content of cellular samples from 54 patients affected by breast carcinomas and 20 affected by benign breast lesions (including 11 fibroadenomas) was measured by flow cytometry. All normal tissue samples and 17/20 (85%) specimens from benign lesions exhibited a cytometrically diploid DNA distribution, 3/20 (15%) benign lesions an abnormal DNA content, and 35/54 (65%) carcinomas at least one aneuploid cell subpopulation. Furthermore, 9/54 (17%) tumors were characterized by the presence of more than one aneuploid cell subpopulation. The results also indicate that flow cytometry can be used to recognize lymph nodes infiltrated by aneuploid cells. Statistically significant correlations were evidenced between the occurrence of aneuploidy or the ploidy level measured as DNA index and the nodal infiltration status. The percentage of S cells can also be extracted from DNA content distribution histograms. Statistically significant differences (p < 0.01) were also observed for the percentage of S cells between normal tissues (6.2±3.2 SD) and benign lesions (11.1±6.6 SD), normal tissues (6.2 ± 3.2 SD) and aneuploid tumors (19.7 ± 10.3 SD), benign lesions (11.1 ± 6.6 SD) and aneuploid tumors (19.7 ± 10.3 SD), and diploid (7.9 ± 4.0 SD) and aneuploid tumors (19.7 ± 10.3 SD).

DNA content in human colon cancer and non-neoplastic adjacent mucosa

1995 ◽  
Vol 10 (1) ◽  
pp. 11-16 ◽  
Author(s):  
G. Saccani Jotti ◽  
M. Fontanesi ◽  
N. Orsi ◽  
L. Sarli ◽  
N. Pietra ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Human Colon ◽  
Human Colon Cancer ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Adjacent Mucosa ◽  
Aneuploid Tumor ◽  
The Relationship

DNA content was determined by flow cytometry in a series of 51 paired fresh tissue samples of primary colorectal carcinomas and the respective non-neoplastic adjacent mucosa in order to assess the relationship between DNA ploidy and the most commonly used prognostic factors. Aneuploidy was observed in 70.6% of the tumors and more than one aneuploid peak was present in 3.9%. Aneuploid tumor frequency was higher in left (93.3%) and right colon (64.7%) cancers than in rectal carcinomas (60.0%), and multiple aneuploid clones were detected more frequently in men than in women and in patients with advanced disease (Dukes stage D). Non-neoplastic mucosa adjacent to aneuploid tumors showed aneuploidy in 4 out of 51 samples (7.8%). The mucosa adjacent to diploid cancers had only diploid characteristics. Polidy did not correlate with histological abnormalities. These findings suggest that DNA content as determined by flow cytometry needs further study with adequate follow-up to evaluate possible correlations with relapse-free and overall survival. Furthermore the aneuploidy of non-neoplastic mucosa provides evidence for a field defect in mucosa adjacent to colorectal cancer and supports the concept that this alteration may be of influence on carcinogenesis.

scholarly journals Penetration of Piperacillin/Tazobactam Into Gynecologic Tissues Following a Single Loading Dose Prior to Hysterectomy

1994 ◽  
Vol 2 (1) ◽  
pp. 20-24
Author(s):  
William F. O'Brien ◽  
Shelley George
Keyword(s):  
Intravenous Administration ◽  
Genital Tract ◽  
Plasma Concentrations ◽  
Female Genital Tract ◽  
Loading Dose ◽  
Effective Agent ◽  
Tissue Concentrations ◽  
Tissue Samples ◽  
Single Intravenous Administration ◽  
Female Genital

Objective:This study was undertaken to determine plasma and pelvic tissue concentrations of piperacillin/tazobactam following a single intravenous administration.Methods:Plasma and tissue samples were obtained following administration of 3 g of piperacillin and 0.375 g of tazobactam. Concentrations of these agents were determined by chromatography.Results:Plasma concentrations (mean ±SD) for piperacillin/tazobactam were 208 ± 87/27 ± 8 and 95 ± 53/14 ± 2, respectively, at 30 and 60 min after the start of the infusion. The 8:1 ratio between piperacillin and tazobactam in plasma remained constant for up to 96 min. Tissue concentrations yielded a broader range, but each agent achieved significant penetration in each of the tissues studied.Conclusions:Piperacillin/tazobactam demonstrates favorable penetration in female genital tract tissues and Should be considered a potentially effective agent for female pelvic infections.

scholarly journals Multichromatic phenotyping of HER receptor coexpression in breast tumor tissue samples using flow cytometry-Possibilities and limitations

2010 ◽  
Vol 77A (4) ◽  
pp. 387-398 ◽  
Author(s):  
Arabel Vollmann-Zwerenz ◽  
Simone Diermeier-Daucher ◽  
Anja Kathrin Wege ◽  
Andrea Sassen ◽  
Elisabeth Schmidt-Brücken ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Breast Tumor ◽  
Tumor Tissue ◽  
Tissue Samples ◽  
Breast Tumor Tissue

scholarly journals Female genital tuberculosis: A disease seen again in Europe

2007 ◽  
Vol 64 (12) ◽  
pp. 855-858 ◽  
Author(s):  
Dragica Pesut ◽  
Jelena Stojsic
Keyword(s):  
Abdominal Mass ◽  
Reproductive Organs ◽  
Medical Problem ◽  
Mycobacterium Tuberculosis Infection ◽  
Tissue Samples ◽  
Younger Women ◽  
Antituberculosis Treatment ◽  
Immunodeficiency Virus ◽  
Female Genital Tuberculosis ◽  
Female Genital

Background. Dramatic worsening of epidemiological situation with tuberculosis (TB) in the world has made extra- pulmonary tuberculosis actual again. Female genital TB is very rare, but each case still remains a serious medical problem. Case report. A 23-year-old human immunodeficiency virus (HIV) seronegative woman with two-month duration amenorrhea underwent surgery due to lower abdominal mass simulating a left ovary carcinoma, suggested by ultrasound examinations. During sampling, we found a mass of round, up to 3 mm, necrotizing nodules, diffuse in the uterus, ovarian and tubarial surfaces, in cervical and endometrial mucosa, and even in myometrium and fat omental tissue. No tumor mass was found. Microscopically, the tissue samples from all reproductive organs and omentum contained numerous tuberculous caseating granulomas. Mycobacteria were identified by Ziehl-Neelsen method. Antituberculosis treatment had been completed. Conclusion. In the differential diagnosis of an ovarian tumor and ascites TB should always be considered. It should also be suspected in recent Mycobacterium tuberculosis infection in younger women with amenorrhea, either HIV-seropositive or not.

scholarly journals Comparative DNA Flow Cytometric Study of Primary Intraocular and Central Nervous System Lymphomas

2020 ◽  
Vol 8 (2) ◽  
Author(s):  
Teresa Martinu ◽  
Claudia P. Correia ◽  
Andersson M. Figueiredo ◽  
Miguel N. Burnier Jr
Keyword(s):  
Flow Cytometry ◽  
Dna Content ◽  
Primary Cns Lymphoma ◽  
Spatial Location ◽  
Intraocular Lymphoma ◽  
Dna Flow Cytometry ◽  
Cns Lymphoma ◽  
Primary Intraocular Lymphoma ◽  
Human Tonsil ◽  
Tissue Samples

Primary intraocular lymphoma is generally considered as a subset of primary CNS lymphoma. This study attempts to show that they may in fact represent distinct entities by comparing their respective proliferation rates using DNA flow cytometry. Four samples of primary intraocular lymphoma and seven samples of primary CNS lymphoma were analyzed, all from paraffin-embedded tissue. All tumors were of the large B-cell type. A normal human tonsil sample was used as a control. Tissue samples were analyzed by DNA flow cytometry, which is a precise and objective method to measure DNA content and cell proliferation of a tumor. S-phase fraction (SPF) and DNA content were measured for each sample. The average SPF for primary intraocular lymphoma was significantly higher than that of primary CNS lymphoma, 23.8 (range: 18.9 to 29.6) versus 15.1 (range: 1.1 to 25.1) respectively. Of the 11 tumors analyzed, 2 brain tumors were aneuploid and 1 eye tumor was peridiploid. All other tumors were diploid. Thus, no significant pattern was detected in the DNA content of the tumors. This lack of clinical significance of tumor aneuploidy is consistent with data reported in the literature. The results of this study indicate that primary intraocular lymphoma is more aggressive and of higher grade than primary CNS lymphoma. The different proliferation rates of intraocular and CNS lymphomas may be explained by either their different spatial location or a distinct genetic composition, the latter reinforcing the hypothesis that the two are fundamentally different entities

scholarly journals Exponential fluorescent amplification of individual RNAs using clampFISH probes

2017 ◽  
Author(s):  
Sara H. Rouhanifard ◽  
Ian A. Mellis ◽  
Margaret Dunagin ◽  
Sareh Bayatpour ◽  
Orsolya Symmons ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Single Molecule ◽  
Signal Amplification ◽  
Modular Design ◽  
High Specificity ◽  
High Gain ◽  
Fluorescent Detection ◽  
Tissue Samples ◽  
Locking Mechanism

AbstractNon-enzymatic, high-gain signal amplification methods with single-cell, single-molecule resolution are in great need. We present click-amplifying FISH (clampFISH) for the fluorescent detection of RNA that combines the specificity of oligonucleotides with bioorthogonal click chemistry in order to achieve high specificity and extremely high-gain (>400x) signal amplification. We show that clampFISH signal enables detection with low magnification microscopy and separation of cells by RNA levels via flow cytometry. Additionally, we show that the modular design of clampFISH probes enables multiplexing, that the locking mechanism prevents probe detachment in expansion microscopy, and that clampFISH works in tissue samples.

scholarly journals PPAR-γAgonist Alleviates Liver and Spleen Pathology via Inducing Treg Cells duringSchistosoma japonicumInfection

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Yuxiao Zhu ◽  
Yangyue Ni ◽  
Ran Liu ◽  
Min Hou ◽  
Bingya Yang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Schistosoma Japonicum ◽  
Cellular Proliferation ◽  
Inflammatory Responses ◽  
Treg Cells ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Tissue Samples

Background. Peroxisome proliferator-activated receptor- (PPAR-)γplays critical roles in human metabolic disorders and has recently been implicated as a regulator of cellular proliferation and inflammatory responses. Regulatory T cells (Tregs), which express high levels of PPAR-γprotein, have the ability to maintain immune tolerance to self-antigens and regulate immune response toSchistosomainfection. However, mechanisms involved in the resolution of these responses are elusive.Methods. Liver and spleen tissue samples inSchistosoma japonicum-infected mice after administration of pioglitazone (a PPAR-γagonist) were collected. The hepatic and splenic pathologies were detected by H&E and Masson staining. The percentages of Th1/2 and Treg cells in the liver and spleen of each mouse were determined using flow cytometry. Levels of gene expression of PPAR-γand Foxp3 in tissues or cells were determined using real-time PCR (RT-PCR). Macrophages were treated with pioglitazonein vitroor cocultured with normal purified CD4+T cells for detecting Treg cells by flow cytometry. The interactions of PPAR-γwith Foxp3 in CD4+T cells were detected by coimmunoprecipitation.Results. Administration of pioglitazone resulted in the prevention of the development of hepatic and splenic pathologies. Activation of PPAR-γby pioglitazone resulted in increased percentages of CD4+CD25+Foxp3+Treg cells and decreased percentages of CD3+CD4+IFN-γ+and CD3+CD4+IL-4+cells in the liver and spleen ofSchistosoma japonicum-infected mice. In addition, the PPAR-γagonist can induce Treg cellsin vitrodirectly or by modulating the macrophage’s function indirectly. Furthermore, through interaction with Foxp3 in CD4+T cells, the PPAR-γagonist can promote the expression of Foxp3; however, the inhibitor of PPAR-γweakened the expression of Foxp3 by modifying the coexpression of Foxp3 and PPAR-γ.Conclusions. Our study reveals a previously unrecognized role for PPAR-γ/Foxp3 signaling in regulating the immunopathology that occurs duringSchistosomainfection through induction of Treg cells.

scholarly journals A Simple and Effective Flow Cytometry-Based Method for Identification and Quantification of Tissue Infiltrated Leukocyte Subpopulations in a Mouse Model of Peripheral Arterial Disease

2020 ◽  
Vol 21 (10) ◽  
pp. 3593 ◽  
Author(s):  
Konda Kumaraswami ◽  
Natallia Salei ◽  
Sebastian Beck ◽  
Stephan Rambichler ◽  
Anna-Kristina Kluever ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Arterial Disease ◽  
Tissue Samples ◽  
Collateral Arteries ◽  
Optimized Protocol ◽  
Adductor Muscles ◽  
Protocol Flow ◽  
Peripheral Arterial ◽  
Perivascular Infiltration ◽  
Detailed Protocol

Arteriogenesis, the growth of a natural bypass from pre-existing arteriolar collaterals, is an endogenous mechanism to compensate for the loss of an artery. Mechanistically, this process relies on a locally and temporally restricted perivascular infiltration of leukocyte subpopulations, which mediate arteriogenesis by supplying growth factors and cytokines. Currently, the state-of-the-art method to identify and quantify these leukocyte subpopulations in mouse models is immunohistology. However, this is a time consuming procedure. Here, we aimed to develop an optimized protocol to identify and quantify leukocyte subpopulations by means of flow cytometry in adductor muscles containing growing collateral arteries. For that purpose, adductor muscles of murine hindlimbs were isolated at day one and three after induction of arteriogenesis, enzymatically digested, and infiltrated leukocyte subpopulations were identified and quantified by flow cytometry, as exemplary shown for neutrophils and macrophages (defined as CD45+/CD11b+/Ly6G+ and CD45+/CD11b+/F4/80+ cells, respectively). In summary, we show that flow cytometry is a suitable method to identify and quantify leukocyte subpopulations in muscle tissue, and provide a detailed protocol. Flow cytometry constitutes a timesaving tool compared to histology, which might be used in addition for precise localization of leukocytes in tissue samples.

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