scholarly journals Quantification of telomere features in tumor tissue sections by an automated 3D imaging-based workflow

2016 ◽  
Author(s):  
Manuel Gunkel ◽  
Inn Chung ◽  
Stefan Wörz ◽  
Katharina I. Deeg ◽  
Ronald Simon ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence Microscopy ◽  
Fluorescence In Situ Hybridization ◽  
Length Distribution ◽  
Nuclear Bodies ◽  
Telomere Maintenance ◽  
Proliferative Potential ◽  
Tissue Sections ◽  
Pml Nuclear Bodies

AbstractThe microscopic analysis of telomere features provides a wealth of information on the mechanism by which tumor cells maintain their unlimited proliferative potential. Accordingly, the analysis of telomeres in tissue sections of patient tumor samples provides can be exploited to obtain diagnostic information and to define tumor subgroups. In many instances, however, analysis of the image data is conducted by manual inspection of 2D images at relatively low resolution for only a small part of the sample. As the telomere feature signal distribution is frequently heterogeneous, this approach is prone to a biased selection of the information present in the image and lacks subcellular details. Here we address these issues by using an automated high-resolution imaging and analysis workflow that quantifies individual telomere features on tissue sections for a large number of cells. The approach is particularly suited to assess telomere heterogeneity and low abundant cellular sub-populations with distinct telomere characteristics in a reproducible manner. It comprises the integration of multi-color fluorescence in situ hybridization, immunofluorescence and DNA staining with targeted automated 3D fluorescence microscopy and image analysis. We apply our method to telomeres in glioblastoma and prostate cancer samples, and describe how the imaging data can be used to derive statistically reliable information on telomere length distribution or colocalization with PML nuclear bodies. We anticipate that relating this approach to clinical outcome data will prove to be valuable for pretherapeutic patient stratification.Abbreviations3D-TIM3D targeted imagingALTalternative lengthening of telomeresAPBALT-associated PML-NBCLSMconfocal laser scanning fluorescence microscopyECTRextrachromosomal telomeric repeatFFPEformalin-fixed, paraffin-embeddedFISHfluorescence in situ hybridizationIFImmunofluorescencepedGBMpediatric glioblastomaPMLpromyelocytic leukemiaPML-NBPML nuclear bodyPNApeptide nucleic acidROIregion of interestTMAtissue microarrayTMMtelomere maintenance mechanismSMLMsingle molecule localization microscopy

Fluorescence in situ hybridization for the identification of Treponema pallidum in tissue sections

2015 ◽  
Vol 305 (7) ◽  
pp. 709-718 ◽  
Author(s):  
Annett Petrich ◽  
Pablo Rojas ◽  
Julia Schulze ◽  
Christoph Loddenkemper ◽  
Lorenzo Giacani ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Treponema Pallidum ◽  
Tissue Sections

Detection of t(14;18) in follicular lymphoma by dual-color fluorescence in situ hybridization on paraffin-embedded tissue sections

2004 ◽  
Vol 150 (1) ◽  
pp. 22-26 ◽  
Author(s):  
Yosuke Matsumoto ◽  
Kenichi Nomura ◽  
Sawako Matsumoto ◽  
Kyoji Ueda ◽  
Mitsushige Nakao ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Follicular Lymphoma ◽  
Fluorescence In Situ Hybridization ◽  
Tissue Sections ◽  
Dual Color

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 Correction to: Use of multicolor fluorescence in situ hybridization to detect deletions in clinical tissue sections

2018 ◽  
Vol 98 (6) ◽  
pp. 839-839 ◽  
Author(s):  
Maisa Yoshimoto ◽  
Olga Ludkovski ◽  
Jennifer Good ◽  
Ciro Pereira ◽  
Robert J. Gooding ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Tissue Sections ◽  
Multicolor Fluorescence

Telomere analysis of platyhelminths and acanthocephalans by FISH and Southern hybridization

Genome ◽  
2009 ◽  
Vol 52 (11) ◽  
pp. 897-903 ◽  
Author(s):  
Marta Bombarová ◽  
Magda Vítková ◽  
Marta Špakulová ◽  
Božena Koubková
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Southern Hybridization ◽  
Telomeric Repeat ◽  
Telomere Maintenance ◽  
Repeat Motif ◽  
Pomphorhynchus Laevis ◽  
Nippotaenia Mogurndae ◽  
Parasitic Worms

We examined the composition of telomeres in chromosomes of parasitic worms, representatives of the flatworm groups Monogenea and Cestoda (Platyhelminthes), and thorny-headed worms (Syndermata: Acanthocephala) by fluorescence in situ hybridization (FISH) with different telomeric repeat probes. Our results show that the (TTAGGG)n sequence, supposed to be the ancestral telomeric repeat motif of Metazoa, is conserved in Monogenea ( Paradiplozoon homoion ) and Cestoda ( Caryophyllaeus laticeps , Caryophyllaeides fennica , and Nippotaenia mogurndae ) but not in Acanthocephala ( Pomphorhynchus laevis and Pomphorhynchus tereticollis ). In the Pomphorhynchus species, no hybridization signals were obtained with the “nematode” (TTAGGC)n, “arthropod” (TTAGG)n, and bdelloid (TGTGGG)n telomeric probes using FISH with their chromosomes and Southern hybridization with P. laevis DNA. Therefore, we suggest that parasitic Acanthocephala have evolved yet unknown telomeric repeat motifs or different mechanisms of telomere maintenance.

scholarly journals Very short telomere length by flow fluorescence in situ hybridization identifies patients with dyskeratosis congenita

Blood ◽  
2007 ◽  
Vol 110 (5) ◽  
pp. 1439-1447 ◽  
Author(s):  
Blanche P. Alter ◽  
Gabriela M. Baerlocher ◽  
Sharon A. Savage ◽  
Stephen J. Chanock ◽  
Babette B. Weksler ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Telomere Length ◽  
Bone Marrow Failure ◽  
Correct Diagnosis ◽  
Dyskeratosis Congenita ◽  
Telomere Maintenance

Abstract Dyskeratosis congenita (DC) is an inherited bone marrow failure syndrome in which the known susceptibility genes (DKC1, TERC, and TERT) belong to the telomere maintenance pathway; patients with DC have very short telomeres. We used multicolor flow fluorescence in situ hybridization analysis of median telomere length in total blood leukocytes, granulocytes, lymphocytes, and several lymphocyte subsets to confirm the diagnosis of DC, distinguish patients with DC from unaffected family members, identify clinically silent DC carriers, and discriminate between patients with DC and those with other bone marrow failure disorders. We defined “very short” telomeres as below the first percentile measured among 400 healthy control subjects over the entire age range. Diagnostic sensitivity and specificity of very short telomeres for DC were more than 90% for total lymphocytes, CD45RA+/CD20− naive T cells, and CD20+ B cells. Granulocyte and total leukocyte assays were not specific; CD45RA− memory T cells and CD57+ NK/NKT were not sensitive. We observed very short telomeres in a clinically normal family member who subsequently developed DC. We propose adding leukocyte subset flow fluorescence in situ hybridization telomere length measurement to the evaluation of patients and families suspected to have DC, because the correct diagnosis will substantially affect patient management.

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