Basic genetics and patterns of inheritance
Hung N. Winn, Frank A. Chervenak, Roberto Romero in Clinical Maternal-Fetal Medicine Online, 2021
Fluorescence in situ hybridization (FISH) is a technique in which DNA probes attached to fluorescent dyes are hybridized to specific chromosome regions from patient samples, either interphase cells or metaphase chromosome spreads, most often from lymphocytes or amniocytes. The chromosomes are then visualized by fluorescent microscopy. Probes can be designed to hybridize to whole chromosomes, specific chromosome segments of interest, centromeres, or telomeres. FISH is widely used for the diagnosis of suspected recognizable microdeletion syndromes (Fig. 4). Multicolor FISH is useful for analyzing submicroscopic structural rearrangements undetectable by classic cytogenetic techniques or for identifying marker chromosomes. FISH is also used for rapid screening for aneuploidy including trisomy 13, 18, or 21 and abnormalities of X and Y.
Methods for Genetic Testing II
Peter G. Shields in Cancer Risk Assessment, 2005
Fluorescence in situ hybridization has several advantages over conventional cytogenetics, including selectivity of specific DNA probes, multiple color labeling, sensitivity of detection, and speed of microscopic analysis. Interphase FISH, in particular, offers several advantages over classical cytoge-netics (21). First, interphase FISH allows analysis of non-dividing cells. Second, a much larger number of cells, at least 1000 or more, may be analyzed. Third, the detection of aneuploidy is facilitated by simply counting the number of labeled regions representing a particular chromosome of interest within the isolated interphase nucleus. By contrast, metaphase FISH can readily detect structural rearrangements in addition to aneuploidy. Furthermore, because metaphase FISH, like classical cytogenetics, analyzes dividing cells, the results from these two methods may be directly compared. A number of studies have determined that FISH is both more sensitive and convenient than classical cytogenetics (22–24). Therefore, FISH appears to be the more suitable method for large-scale population biomonitoring
Molecular Biology
John C Watkinson, Raymond W Clarke, Louise Jayne Clark, Adam J Donne, R James A England, Hisham M Mehanna, Gerald William McGarry, Sean Carrie in Basic Sciences Endocrine Surgery Rhinology, 2018
Hybridization to target DNA in cells, using fluorescence detection, is known as fluorescence in situ hybridization (FISH). Fluorescence in situ hybridization allows the analysis of copy number of a known specific DNA sequence within intact nuclei. In reticuloendothelial malignancies and solid tumour-derived cell lines, the use of both single-copy probes and centromere alpha-satellite repeat probes on metaphase preparations has enhanced and refined classical karyotyping. Interphase FISH has been applied to solid tumour sections to assess the copy number of a known sequence in breast, prostate, bladder, brain, lung and head and neck tumours.
MicroRNA-122-5p ameliorates tubular injury in diabetic nephropathy via FIH-1/HIF-1α pathway
Published in Renal Failure, 2022
Li Cheng, Xinying Qiu, Liyu He, Li Liu
Fluorescence in situ hybridization (FISH) was performed according to the manufacturer's instructions. Briefly, kidneys were harvested from control and STZ-treated mice to prepare 4-micron paraffin section. The sections were treated with 20 μg/ml proteinase K for permeabilization, and then incubated with pre-hybridization solution at 78 °C for 1 h. Remove pre-hybridization solution and add digoxigenin-labeled mmu-miR-122-5p LNA probe over night at 37 °C. At the second day, after wash, bovine serum albumin (BSA) was added for blocking. Then the anti-digoxigenin-HRP was used at 37 °C for 1 h. CY3-TSA and DAPI assay were used to indicate the positive areas and cell nucleus respectively. The images were acquired from a fluorescence microscope and the representative figures were exhibited.
Microsatellite Instability and Promoter Hypermethylation of DNA repair genes in Hematologic Malignancies: a forthcoming direction toward diagnostics
Published in Hematology, 2018
Priyanjali Bhattacharya, Trupti N. Patel
Since the discovery of trisomy 21 in 1959, clinicians are following cytogenetic tests to date to determine the varying effect of non-curable diseases, including malignancies [11]. In hematology, a conventional cytogenetic analysis helps in identifying structural and/or numerical anomalies in chromosomes, which is formally known as karyotyping. Fluorescent in situ hybridization is another technique which requires bone marrow or peripheral blood samples, but inadequate number of DNA probes and cost limit its performance and use. Spectral karyotyping, even though not enlisted in regular practice, uses unique probes/dyes to paint the 24 pairs of chromosomes. Studies have shown that there are many cases where conventional cytogenetics has missed out certain subtle micro anomalies. In malignancies, chromosomal imbalances cannot be detected using array comparative genomic hybridization, but it is likely to be used for tumor classification and progression. A day-to-day use of cytogenetic markers is more assimilated into the diagnostic, prognostic and therapeutic workgroup of hematologic malignancies rather than any other field of oncology. However, it eventually omits out certain molecular level alterations and thus fails to understand the origin and progression of disease which is important for further establishment of novel therapeutic targets [12]. Thus, it is best to enhance the arena of diagnosis in hematologic malignancies to ponder over certain unaccountable cases.
Approaching complexity: systems biology and ms-based techniques to address immune signaling
Published in Expert Review of Proteomics, 2020
Joseph Gillen, Caleb Bridgwater, Aleksandra Nita-Lazar
When investigating molecules in situ, the question of molecular origin becomes apparent, especially when investigating the interface between host and microbe. Geier et al. address this problem by investigating the metabolomic phenotypes of host and microbe cells in situ. The effective rastering and high resolution of the current MALDI techniques, as previously discussed by Kompauer et al. [52] meant that there was minimal sample damage after MALDI-MSI. This translates to sample integrity for further downstream experimentation. Fluorescent in situ hybridization (FISH) is a method of fluorescently labeling complementary nucleic acid sequences on cytochromes. Geier et al. developed a pipeline of high resolution MALDI-MSI and subsequent FISH microscopy on the same biological sample. This image overlay allows for the metabolomic topography to be visualized and host or microbe metabolites to be identified at a resolution of 3 µm when optimized [53]. This pipeline would have many applications in infectious disease and immunology research beyond the exemplary host-microbe symbiosis described in their study.
Related Knowledge Centers
- Cytogenetics
- DNA
- DNA Sequencing
- Nucleic Acid
- Messenger Rna
- Chromosome
- Hybridization Probe
- Complementarity
- Fluorescence Microscope
- Genetic Counseling