TY - JOUR. T1 - High-speed dual color fluorescence lifetime endomicroscopy for highly-multiplexed pulmonary diagnostic applications and detection of labeled bacteria. AU - Pedretti, Ettore. AU - Tanner, Michael G.. AU - Choudhary, Tushar R. AU - Krstajic, Nikola. AU - Megia-Fernandez, Alicia. AU - Henderson, Robert K.. AU - Bradley, Mark. AU - Thomson, Robert R.. AU - Girkin, John M.. AU - Dhaliwal, Kevin. AU - Dalgarno, Paul A.. PY - 2019/1/1. Y1 - 2019/1/1. N2 - We present a dual-color laser scanning endomicroscope capable of fluorescence lifetime endomicroscopy at one frame per second (FPS). The scanning system uses a coherent imaging fiber with 30,000 cores. High-speed lifetime imaging is achieved by distributing the signal over an array of 1024 parallel single-photon avalanche diode detectors (SPADs), minimizing detection dead-time maximizing the number of photons detected per excitation pulse without photon pile-up to achieve the high frame rate. This also enables dual color fluorescence ...

RESULTS. Fluorescence in-situ hybridisation detected 558 (9.5%) patients with chromosomal abnormalities. Abnormal ultrasounds (70%) and maternal serum screens (21%) were the most indicative of chromosomal abnormalities. When comparing fluorescence in-situ hybridisation data with karyotype results for the five chromosomes of interest, the sensitivity and specificity were 99.3% and 99.9%, respectively. When comparing fluorescence in-situ hybridisation data with karyotype results for all chromosomes, the sensitivity decreased to 86.8%, whereas the specificity remained at 99.9%. Of 643 cases with karyotype abnormalities, 85 were fluorescence in-situ hybridisation-negative (false negative rate, 13.2%), which included structural rearrangements, chromosome mosaicism, and other trisomies. Despite abnormal ultrasound indications, fluorescence in-situ hybridisation missed 32 cases which included structural rearrangements, mosaicisms, and other trisomies ...

16120DNAArtificial SequenceYM-1 Forward Primer 1tggaattggt gcccctacaa 20220DNAArtificial SequenceYM-1 Reverse Primer 2aacttgcact gtgtatattg 20318DNAArtificial SequenceYM-2 Forward Primer 3aacctcagac attcatta 18421DNAArtificial SequenceYM-2 Reverse Primer 4tggtccttcc agtaggtaat a 21520DNAArtificial SequenceYM-3 Forward Primer 5tataaatctc catttgacac 20620DNAArtificial SequenceYM-3 Reverse Primer 6cctaatttat tgtccttgac 20728DNAArtificial SequenceAMCase Forward Primer 7atctgcagtg gacacacctt catcctga 28828DNAArtificial SequenceAMCase Reverse Primer 8atgaattcaa caagccctgc ttgacaat 28922DNAArtificial SequenceYM Antisense In Situ Hybridization Probe 9tcctcgagac ccagggtact gc 221024DNAArtificial SequenceYM Sense In Situ Hybridization Probe 10tatctagagg atcttcctac cagc 241129DNAArtificial SequenceAMCase Antisense In Situ Hybridization Probe 11tcgctcgaga acaagccctg cttgacaat 291228DNAArtificial SequenceAMCase sense In Situ Hybridization Probe 12gctctagatg gacacacctt catcctga 281319PRTArtificial ...

TY - JOUR. T1 - Chromosome microdissection. T2 - A brief overview. AU - Cannizzaro, L. A.. PY - 1996/1/1. Y1 - 1996/1/1. N2 - Chromosome microdissection arose as a means of facilitating long range physical mapping of chromosome regions involved in either a genetic or malignant disorder. However, with the rapid development of improved techniques for mapping and sequencing the human genome, microdissection is considered by many investigators to be a cumbersome and time consuming procedure. Nonetheless, based on the impressive number of informative diagnostic DNA markers that are now available as a result of this technology, microdissection still must be considered one of the most rapid and direct methods available for generating new DNA markers from any chromosome region, irrespective of its sequence composition. In addition, it remains an important means to dissect DNA markers from any organism, eukaryotic and prokaryotic, and has resulted in generating disease associated DNA sequences from both ...

TY - JOUR. T1 - Molecular cytogenetic study and chromosome abnormalities in Alzheimers disease. Protease-inhibitory peptides and amyloid beta protein in Alzheimers disease. AU - Ito, H.. AU - Kitaguchi, N.. PY - 1988/7/1. Y1 - 1988/7/1. UR - http://www.scopus.com/inward/record.url?scp=0024046766&partnerID=8YFLogxK. UR - http://www.scopus.com/inward/citedby.url?scp=0024046766&partnerID=8YFLogxK. M3 - Article. C2 - 3065525. AN - SCOPUS:0024046766. VL - 46. SP - 1514. EP - 1520. JO - Nippon rinsho. Japanese journal of clinical medicine. JF - Nippon rinsho. Japanese journal of clinical medicine. SN - 0047-1852. IS - 7. ER - ...

TY - JOUR. T1 - Eigenanalysis of DAPI-stained chromosomes. T2 - Tools and strategies toward computer-assisted analysis of FISH experiments. AU - Knapp, R. D.. AU - Smith, L. C.. AU - Baldini, A.. PY - 1995. Y1 - 1995. N2 - The fluorescent dye 4,6-diamidino-2-phenylindole (DAPI) is widely used as a chromosome counterstain in fluorescence in situ hybridization (FISH) studies. It produces a Q-banding pattern that allows for both chromosome identification and the assignment of molecular probes to specific chromosome bands. Using a statistical procedure based on eigenanalysis, we have extracted features from digital images of DAPI-stained chromosomes and constructed prototypes of each of the 24 human chromosomes. The features of these prototypes are directly proportional, in intensity profile and band location, to those of real chromosomes. The prototypes intensity profile can be translated into cytogenetic bands to provide a computer-based strategy for chromosome mapping and analysis amenable to ...

Before Its News). Global FISH Probe Market 2016-2020 Order This Report by calling BigMarketResearch.com at +1-971-202-1575.. About FISH Probe. FISH probe is a molecular diagnostic technique that utilizes labeled DNA probes to either detect or confirm gene or abnormalities in the chromosome. Analysts forecast the global fish probe market to grow at a CAGR of 7.34% during the period 2016-2020.. Covered in this report. The report covers the present scenario and the growth prospects of the global fish probe market for 2016-2020. To calculate the market size, we use the revenue generated from the sale of FISH probes.. Get sample copy of report @ https://goo.gl/CQ1yEO. The market is divided into the following segments based on geography:. • Americas. • APAC. • Europe. Global Fish Probe Market 2016-2020, has been prepared based on an in-depth market analysis with inputs from industry experts. The report covers the market landscape and its growth prospects over the coming years. The report also ...

Headline: Bitcoin & Blockchain Searches Exceed Trump! Blockchain Stocks Are Next!. The FISH Probe Industry report covers the present scenario and the growth prospects of the FISH Probe Market for 2016-2020. FISH Probe Market report focuses on the major drivers and restraints for the key players. To calculate the market size, the report considers both the direct revenue and the indirect revenue of the vendors. The global fish probe market to grow at a CAGR of 7.34% during the period 2016-2020.. FISH probe is a molecular diagnostic technique that utilizes labeled DNA probes to either detect or confirm gene or abnormalities in the chromosome.. Browse more detail information about FISH Probe Market Report at: http://www.absolutereports.com/global-fish-probe-market-2016-2020-10351017. The report covers the market landscape and its growth prospects over the coming years. The report also includes a discussion of the key vendors operating in this market.. Key Vendors in FISH Probe Market:. • Agilent ...

Fluorescence in situ hybridization (FISH) is a valuable cytogenetic technique for the detection and localization of specific DNA or RNA sequences on chromosomes. Typically, this technique can be used to define spatiotemporal expression patterns of target genes. Using fluorescent DNA probes with sequences complementary to the location of the chromosome of interest, gene expression patterns can be easily detected by fluorescence microscopy and further quantitatively analyzed. In addition, improved FISH analysis also allows simultaneous observation of multiple genes by labeling different sequence fragments with different fluorophores.. In plant systems, FISH analysis has been used in a variety of crops, including wheat, rye, cucumber and melon. Based on the information on Lifeasibles official website, Lifeasibles services cover every step of the FISH assay in the plant system:. • Probe designing and construction: aside from commercially available probes, Lifeasible also provides customized ...

Order Gene Specific Fluorescence In Situ Hybridization (FISH) Probes that have been designed and optimized for the gene of your specific research.

Survival rates for lung cancer are low because patients have disseminated disease at diagnosis; therefore tests for early diagnosis are highly desirable. This pilot study investigated occurrence of chromosomal aneusomy in sputum from a 33 case-control cohort matched on age, gender, and date of sample collection. Subjects had chronic obstructive pulmonary disease and , or = 30 pack-years of tobacco use, and aneusomy was tested using a multi-target DNA FISH assay (LAVysion, Abbott/Vysis). In specimens collected within 12 months of lung cancer diagnosis, abnormality was more frequent among the 18 cases (41%) than the 17 controls (6%; P = 0.04). Aneusomy had no significant association with cytologic atypia, which might indicate that molecular and morphological changes could be independent markers of tumorigenesis. Combining both tests, abnormality was found in 83% of the cases and 20% of the controls (P = 0.0004) suggesting that FISH may improve the sensitivity of cytologic atypia as a predictor of ...

Human chromosome-specific DNA libraries: Construction and availability. Bio-Technology 4: 537-552, 1986. Fuscoe JC, et al. Construction of fifteen human chromosome-specific DNA libraries from flow-purified chromosomes. Cytogenet. Cell Genet. 43: 79-86, 1986. PubMed: 3780319 Perlman J, Fuscoe JC. Molecular characterization of the purity of seven human chromosome-specific DNA libraries. Cytogenet. Cell Genet. 43: 87-96, 1986. PubMed: 3780320 Deaven LL, et al. Construction of human chromosome-specific DNA libraries from flow sorted chromosomes. Cold Spring Harbor Symp. Quant. Biol. 51: 159-167, 1986. PubMed: 3472712 Fuscoe JC. Human chromosome-specific DNA libraries: use of an oligodeoxynucleotide probe to detect non-recombinants. Gene 52: 291-296, 1987. PubMed: 3609744 Marvin Van Dilla, personal communication ...

hybridization (FISH) is a powerful molecular cytogenetic technique which allows rapid detection of aneuploidies on interphase cells and metaphase spreads. The aim of the present study was to evaluate FISH as a tool in prenatal diagnosis of aneuploidies in high risk pregnancies in an Indian set up. Prenatal diagnosis was carried out in 88 high-risk pregnancies using FISH and cytogenetic analysis. Multicolour commercially available FISH probes specific for chromosomes 13, 18, 21, X and Y were used. Interphase FISH was done on uncultured cells from chorionic villus and amniotic fluid samples. FISH on metaphase spreads was done from cord blood samples. The results of FISH were in conformity with the results of cytogenetic analysis in all the normal and aneuploid cases except in one case of structural chromosomal abnormality. The hybridization efficiency of the 5 probes used for the detection of aneuploidies was 100%. Using these probes FISH assay yielded discrete differences in the signal profiles ...

The purpose of the present study was to determine the effect of a single oral dose of carbendazim (CARB) on the frequencies of numerical chromosome aberrations in sperm and on micronuclei in peripheral blood erythrocytes of rats. Dual colour FISH on

Purpose: This study aimed to search for predictors of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) efficacy in previously treated patients with advanced squamous cell lung carcinoma in which EGFR mutations are very rare.. Experimental Design: EGFR gene copy numbers were assessed by FISH and evaluated as predictors of EGFR-TKI efficacy in 71 patients with advanced squamous cell lung cancer who received gefitinib or erlotinib as a second-line or higher therapy. The tumors were classified into EGFR/FISH-positive (high polysomy/gene amplification) and EGFR/FISH-negative (other) groups.. Results: EGFR/FISH was positive in 19 (26.7%) patients. Only EGFR/FISH positive status was correlated with the EGFR-TKIs response (EGFR/FISH+ vs. EGFR/FISH−, 26.3% vs. 2.0%; P = 0.005). In a multivariate analysis, the risk of progression was lower in EGFR/FISH-positive patients (HR of EGFR/FISH+ vs. EGFR/FISH−, 0.57; P = 0.057) or patients experiencing grade 2 or more rash (HR for rash ...

TY - JOUR. T1 - erb-b2 amplification by fluorescence in situ hybridization in breast cancer specimens read as 2+ in immunohistochemical analysis. AU - Lan, Chieh. AU - Liu, Jacqueline Ming. AU - Liu, Tsang Wu. AU - Hsu, Der Hung. AU - Liang, Shuching. AU - Chen, Jim Ray. AU - Peng, Jacqueline Whang. PY - 2005/7/1. Y1 - 2005/7/1. N2 - We conducted this study to ascertain the prevalence of erb-b2 gene amplification in breast cancer specimens read as 2+ in immunohistochemical analysis. Slides from patients with metastatic or recurrent breast cancer were eligible for fluorescent in situ hybridization (FISH) study if they were read as 2+ immunohistochemically for erb-b2 by a certified pathologist. The PathVysion kit (Vysis, Downers Grove, IL) was used for FISH studies. Amplification of the erb-b2 gene was defined as an erb-b2/CEP17 (chromosome 17 centromere) ratio of 2 or more in 30 tumor cells counted. From May 2003 to June 2004, 221 slides were submitted from 24 hospitals around the island. Of 216 ...

Patient management based on HER2 status in breast carcinoma is an archetypical example of personalized medicine but remains hampered by equivocal testing and intratumoral heterogeneity. We developed a fully automated, quantitative, bright-field in situ hybridization technique (RNAscope), applied it to quantify single-cell HER2 mRNA levels in 132 invasive breast carcinomas, and compared the results with those by real-time quantitative PCR (qPCR) and Food and Drug Administration-approved methods, including fluorescence in situ hybridization (FISH), IHC, chromogenic in situ hybridization, and dual in situ hybridization. Both RNAscope and qPCR were 97.3% concordant with FISH in cases in which FISH results were unequivocal. RNAscope was superior to qPCR in cases with intratumoral heterogeneity or equivocal FISH results. This novel assay may enable ultimate HER2 status resolution as a reflex test for current testing algorithms. Quantitative in situ RNA measurement at the single-cell level may be ...

TAMGeS genotypes all the possible SNPs by performing three SBE reactions (P 1 , P 2 and U) with different combinations of labelled ddNTPs, hybridized on three identical arrays. Other dual-colour approaches request either a distinct array for each of the possible base changes or the use of one labelled allele-specific probe for each SNP [21, 23, 27].. Most of the existing methods, based both on tetra- and dual-colour approaches, do not succeed in genotyping all the SNPs with equal efficiency; SNPs with a too high data loss are simply discarded from the analysis. If many SNPs are analysed, as in large scale studies or wide genome scans, such a loss of data does not usually compromise the overall informative power of the study. On the contrary, in the context of association studies of candidate genes and SNPs, retrieving maximum information is essential. Our approach guarantees acquisition of reliable data also in those cases where SNP genotyping proves to be difficult. Indeed, with the three-array ...

Results. Comparison of sample recoveries for chromosome 3 fluorescence in-situ hybridization assay and mapping array analysis. Of the 59 patients who underwent FNAB, FISH results were obtained in 38 (64%) of the cases. Parallel, pooled aspirates (range; 2-4) from each patient were processed for simultaneous isolation of DNA and RNA, and the nucleic acid recoveries were determined. Where DNA recoveries exceeded 350 ng, samples were determined to be adequate for mapping array analysis. Of the 59 patients who underwent FNAB, 49 (83%) of the cases yielded adequate DNA, ranging 380-3040 ng. Six of these 49 failed to generate adequate probe for microarray due to melanin coprecipitation. Mapping array data were successfully obtained in the remaining 43 cases (73%) of the total cases. Mapping arrays not only provided data in all 38 cases where FISH data were obtained, but also provided data in five patients in whom FISH data were not obtained.. Comparison of findings of chromosome 3 fluorescence in-situ ...

10p Subtelomere(Texas Red) FISH Probe FE0092-20uL Labeled FISH probes for identification of subtelomere aberrations using Fluorescent In Situ Hybridization Tech

Principal Investigator：Taniwaki Masafumi, Project Period (FY)：2016-04-01 - 2019-03-31, Research Category：Grant-in-Aid for Scientific Research (C), Section：一般, Research Field：Hematology

We have developed a procedure for efficient in situ hybridization of bacterial recombinants created with various types of large-insert cloning vectors. Minimal quantities of crude DNA are amplified and labeled during the degenerate-oligonucleotide-primed polymerase chain reaction. The resulting probes generate high-intensity fluorescent hybridization signals on metaphase chromosomes and on interphase nuclei.

LandMark™ broad range prestained protein marker(dual color) is a dual color prestained size marker. It is a mixture of 10 recombinant purified and prestained polypeptides whose molecular weights are well-adjusted ranging from 7,000 to 240,000 Da. It is provided preblended in a ready-to-use formula and no reconstitution or further dilution is necessary before use. A blue and pink chromophore is covalently bound to proteins, and 10 prestained proteins are visible during electrophoresis or electrophoretic transfer from the gel to a membrane. The 25 KDa and 70 KDa proteins labeled with orange chromophore offer easy identification and serve as a landmark. Because coupling of chromophore to the proteins affects their apparent molecular weights in SDS-PAGE, unstained protein standard should be used for accuracy ,95%.. ...

An increasing body of evidence indicates that the spatial positioning of genes in the interphase nucleus is highly relevant for their function (Lanctot et al, 2007; Meaburn & Misteli, 2007; Misteli, 2007). Fluorescence in situ hybridization (FISH) is a powerful technique to map gene loci in the interphase nucleus. Depending on protocol FISH can either detect DNA or RNA. Both methods have limitations. DNA FISH only detects the physical location of a gene, but can not detect gene activity. RNA FISH, on the other hand, detects transcripts, but might miss a significant number of alleles, since not all alleles of a gene are necessarily transcribed simultaneously. The most efficient way to map gene loci and their activity is sequential RNA and DNA FISH. This is an important technique to uncover how gene positioning is linked to activity.. Simultaneous detection of RNA and DNA for a gene locus is non-trivial. Procedures during DNA FISH particularly denaturation of cellular DNA, can cause significant ...

Sharpen your knives and come to attention because class is in session! Join Mike Cruz, manager of Greenpoint Fish & Lobster Wholesale, as he details the best methods for cleaning and preparing just about every fish you could encounter in the kitchen. Not every fillet is made the same and learning the proper technique can elevate your seafood game to the next level. So, if youre ready to learn how to fillet every fish, Mike has you covered and then some.

Labeled FISH probes for identification of subtelomere aberrations using Fluorescent In Situ Hybridization Technique. (Technology) (FE0030) - Products - Abnova

Labeled FISH probes for identification of subtelomere aberrations using Fluorescent In Situ Hybridization Technique. (Technology) (FE0119) - Products - Abnova

The purpose was to evaluate and compare 5 different HER2 genetic assays with different characteristics that could affect the performance to analyze the human epidermal growth factor 2 (HER2) gene copy number under low and high throughput conditions. The study included 108 tissue samples from breast cancer patients with HER2 immunohistochemistry (IHC) results scored as 0/1+, 2+, and 3+. HER2 genetic status was analysed using chromogenic in situ hybridization (CISH) and fluorescence in situ hybridization (FISH). Scoring results were documented through digital image analysis. The cancer region of interest was identified from a serial H&E stained slide following tissue cores were transferred to a tissue microarrays (TMA). When using TMA in a routine flow, all patients will be tested for HER2 status with IHC followed by CISH or FISH, thereby providing individual HER2 results. In conclusion, our results show that the differences between the HER2 genetic assays do not have an effect on the analytic ...

Easy and accurate HER2 testing is essential when considering the prognostic and predictive significance of HER2 in breast cancer. The use of a fully automated, quantitative FISH assay would be helpful to detect HER2 amplification in breast cancer tissue specimens with reduced inter-laboratory variab …

Empire Genomics RNA5-8S3 FISH probe is used to detect translocations of the RNA5-8S3 gene and can be labeled in one of five colors, using standard nick translation protocols. Each probe is sold in 20 test kits (~20 slides - 22x22 mm area) and includes hybridization buffer. Order 5 or more of the RNA5-8S3 FISH probe and save 10%!

The FN1 5 FISH Probe is designed to detect the human FN1 gene 5-region located on chromosome band 2q35. FN1 is also known as CIG, ED-B, FINC, FN, FNZ, GFND,GFND2, LETS or MSF. The FN1 5 FISH probe is labeled with CytoGreen. CytoGreen is a fluorophore with an excitation peak at 495nm and emission peak at 518nm, givin

00290] Karyotype analysis. Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAPS cells were arrested in metaphase by colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37°C in 5% C02. Cells were washed with PBS, treated with trypsin/ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KC 1 solution was added. Cells were incubated for 20 min at 37°C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3-5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3: 1; vol/vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides. For the FISH procedure, mouse chromosome-specific painting probes were combinatorially labeled using seven different fluorochromes and hybridized as previously described (Jentsch et al, ...

00290] Karyotype analysis. Karyotype analysis was performed by Multicolor FISH analysis (M-FISH). Subconfluent STAPS cells were arrested in metaphase by colcemid (final concentration 0.270 μg/ml) to the culture medium for 2.5 h at 37°C in 5% C02. Cells were washed with PBS, treated with trypsin/ethylenediaminetetraacetic acid (EDTA), resuspended into cell medium and centrifuged for 5 min at 1200 rpm. To the cell pellet in 3 ml of PBS, 7 ml of a prewarmed hypotonic 0.0375 M KC 1 solution was added. Cells were incubated for 20 min at 37°C. Cells were centrifuged for 5 min at 1200 rpm and the pellet was resuspended in 3-5 ml of 0.0375 M KC1 solution. The cells were fixed with methanol/acetic acid (3: 1; vol/vol) by gently pipetting. Fixation was performed four times prior to spreading the cells on glass slides. For the FISH procedure, mouse chromosome-specific painting probes were combinatorially labeled using seven different fluorochromes and hybridized as previously described (Jentsch et al, ...

Abnova 16q Subtelomere (DEAC) FISH Probe 1 Set Life Sciences:Biochemicals and Reagents:Fluorescent in situ hybridization (FisH) Reagents

Custom FISH Probes - Creative Bioarray is pleased to announce the introduction of custom labeled Fluorescence in situ Hybridization (FISH) Probes for a range of molecular and cytogenetic applications. What sets us apart in custom labeled probe manufacturing is our ability to consistently offer high quality locus-specific probes that can be customized to meet the specific needs of researchers and clinicians alike.

Cytogenetic Abnormalities with Interphase FISH Method and Clinical Manifestation in Chronic Lymphocytic Leukemia Patients in North-East of ...

Silver-enhanced in-situ hybridization (SISH) is an emerging tool for the determination of the Her-2/neu amplification status in breast cancer. SISH is technically comparable to fluorescence in-situ hybridization (FISH) but does not require a fluorescence microscope for its interpretation. Although recent studies on histologic evaluations of SISH are promising, we aimed to evaluate its performance on 71 cytologic breast cancer specimens with the new combined Her-2/Chr17 probe. Her-2/neu status as routinely determined by FISH was available for all patients. We found SISH signals in cytologic cell blocks and smear specimens easy to evaluate in most cases. Small numbers of tumor cells and difficulties in identifying tumor cells in lymphocyte-rich backgrounds were limiting factors. Her-2/neu status, as determined by Her-2/Chr17 SISH, was basically identical to the results of the corresponding FISH. The discrepancies were mainly owing to the heterogeneity of Her-2/neu amplification in the tumor ...

TY - JOUR. T1 - The effect of nucleobase-specific fluorescence quenching on in situ hybridization with rRNA-targeted oligonucleotide probes. AU - Behrens, Sebastian. AU - Fuchs, Bernhard M.. AU - Amann, Rudolf. PY - 2004/9. Y1 - 2004/9. N2 - Oligonucleotide probes labeled with fluorescent dyes are used in a variety of in situ applications to detect specific DNA or RNA molecules. It has been described that probe fluorescence might be quenched upon hybridization in a sequence specific way. Here, a set of 17 oligonuleotides labeled with 6-carboxyfluorescein was used to examine the relevance of nucleotide specific quenching for fluorescence in situ hybridization (FISH) to whole fixed bacterial cells. Probes quenched upon hybridization to a guanine-rich region of purified RNA in solution were not quenched upon FISH. Among other factors the high protein concentration within cells may prevent quenching of probe fluorescence in situ.. AB - Oligonucleotide probes labeled with fluorescent dyes are used in ...

In S. bulbocastanum, two strong, one intermediate, and one weak FISH signal were detected on four somatic metaphase chromosomes, with all signals located near the centromeric regions (Figure 2, H-J). Hybridization of 2D8 to NOR on S. bulbocastanum chromosomes was also observed (data not shown) but the cross-hybridization signals were not as strong as those observed in USW1. FISH on S. bulbocastanum pachytene chromosomes also yielded four distinct signals (Figure 2L), indicating that the four loci do not pair with one another and are therefore hemizygous. The high resolution pachytene FISH signals confirmed the pericentromeric locations of the 2D8 repeat (Figure 2M). The pachytene regions associated with the FISH signals are brightly stained by DAPI and highly condensed as compared to the distal euchromatic regions.. The 2D8 repeat is homologous to the IGS sequence of potato rDNA: One 5.9-kb 2D8 subclone was completely sequenced. This 5862-bp sequence consists of two diverged monomers of similar ...

Fluorescent proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state are important for innovative microscopy schemes, such as protein tracking, fluorescence resonance energy transfer imaging, sub-diffraction resolution microscopy and others. However, all available monomeric reversibly switchable fluorescent proteins (RSFPs) have similar properties and switching characteristics, thereby limiting their use. Here, we introduce two bright green fluorescent RSFPs, bsDronpa and Padron, generated by extensive mutagenesis of the RSFP Dronpa, with unique absorption and switching characteristics. Whereas bsDronpa features a broad absorption spectrum extending into the UV, Padron displays a switching behavior that is reversed to that of all green fluorescent RSFPs known to date. These two RSFPs enable live-cell fluorescence microscopy with multiple labels using a single detection color, because they can be distinguished by photoswitching. Furthermore, we demonstrate dual-color

The current reversibly switchable fluorescent proteins (RSFPs) can not be multiplexed. Jakobs and colleagues create two RSFPs with novel switching characteristics that can be used simultaneously in fluorescence microscopy experiments using only one detection color. Fluorescent proteins that can be reversibly photoswitched between a fluorescent and a nonfluorescent state are important for innovative microscopy schemes, such as protein tracking1, fluorescence resonance energy transfer imaging2, sub-diffraction resolution microscopy3,4,5,6,7,8,9 and others. However, all available monomeric reversibly switchable fluorescent proteins (RSFPs) have similar properties and switching characteristics10,11,12, thereby limiting their use. Here, we introduce two bright green fluorescent RSFPs, bsDronpa and Padron, generated by extensive mutagenesis of the RSFP Dronpa10, with unique absorption and switching characteristics. Whereas bsDronpa features a broad absorption spectrum extending into the UV, Padron displays a

Background: Various genetic technologies have been employed in the identification of genomic complexity and refinement of prognostic classification of clinically heterogeneous disease of chronic lymphocytic leukemia (CLL). Objective: The present study of interphase cytogenetics and conventional karyotyping was undertaken to perform comprehensive analysis of CLL genetics with an approach to refine early prognostication of disease. Material & Methods: Retrospective analysis by fluorescence in situ hybridization (FISH) was carried out on total 671 patients of CLL at diagnosis between 2008 and 2015. Conventional cytogenetics studies were performed in 50 of 671 patients using CPG Oligonucleotide + IL-2 and TPA (12-O-Tetradecanyl Phorbol 13-acetate) for stimulation of lymphocytes cultures. Results: Interphase cytogenetics could detect recurrent abnormalities such as del(13q14), +12, del(17p13), del(11q22), del(6q23) in 71% of cases. The incidence of del(13q) was higher in Rai stage 0, I, II (p = 0.0005);

Introduction: HER2 gene amplification or overexpression occurs in 15% to 25% of breast cancers and has implications for treatment and prognosis. The most commonly used methods for HER2 testing are fluorescence in situ hybridization (FISH) and immunohistochemistry. FISH is considered to be the reference standard and more accurately predicts response to trastuzumab, but is technically demanding, expensive, and requires specialized equipment. In situ hybridization is required to be eligible for adjuvant treatment with trastuzumab in Australia. Bright-field in situ hybridization is an alternative to FISH and uses a combination of in situ methodology and a peroxidase-mediated chromogenic substrate such as diaminobenzidine [chromogenic in situ hybridization (CISH)] or multimer technology coupled with enzyme metallography [silver-enhanced in situ hybridization (SISH)] to create a marker visible under bright-field microscopy. CISH was introduced into diagnostic testing in Australia in October 2006. SISH ...

We detected comparable levels of asynchronous replication at 22q11.2 in all tested individuals, that is, control individuals and carriers of either translocations or deletions involving the 22q11.2 region. Based on cases with distinguishable chromosomes 22 we show a non-random nature of the asynchronous replication. In all cases where the origin of the structurally abnormal chromosome 22 was known we detected an earlier replication of the paternal alleles. We hypothesise that a non-random asynchronous replication in this region represents a risk factor for the formation of the 22q11.2 deletion by increasing the probability of an initial mispairing of the parental alleles at the highly homologous low-copy repeats. These initial abnormal conformations may lead, later in meiosis, to an unequal meiotic crossover and thus to the 22q11.2 deletion.. The replication timing results presented here were performed on peripheral blood cells. Our hypothesis would imply that the non-random asynchronous ...

indian babe shanaya.com,rui matsushita,xvidioe,Fluorescence in situ hybridization is to perform localization, qualitative and quantitative analysis of intracellular nucleic acids by hybridizing fluorescently labeled nucleic acid probes with nucleic acids in cells or tissues. Fluorescence in situ hybridization probes include NA probes and NA probes, which can detect nucleic acids in animal tissues and plant tissues quickly and with high specificity.

Microbiological diagnosis of sepsis relies primarily on blood culture data. This study compares four diagnostic methods, i.e. those developed by us: nested, multiplex, qPCR (qPCR) and FISH with commercial methods: SeptiFast (Roche) (SF) and BacT/ALERT® 3D blood culture system (bioMérieux). Blood samples were derived from adult patients with clinical symptoms of sepsis, according to SIRS criteria, hospitalized in the Intensive Care Unit. Using qPCR, FISH, SF, and culture, microbial presence was found in 71.8%, 29.6%, 25.3%, and 36.6% of samples, respectively. It was demonstrated that qPCR was significantly more likely to detect microorganisms than the remaining methods; qPCR confirmed the results obtained with the SF kit in all cases wherein bacteria were detected with simultaneous confirmation of Gram-typing. All data collected through the FISH method were corroborated by qPCR. The qPCR and FISH methods described in this study may constitute alternatives to blood culture and to the few existing

TY - JOUR. T1 - Chromosomal gains measured in cytology samples from women with abnormal cervical cancer screening results. AU - Luhn, Patricia. AU - Houldsworth, Jane. AU - Cahill, Lynnette. AU - Schiffman, Mark. AU - Castle, Philip E.. AU - Zuna, Rosemary E.. AU - Dunn, S. Terence. AU - Gold, Michael A.. AU - Walker, Joan. AU - Wentzensen, Nicolas. PY - 2013/9/1. Y1 - 2013/9/1. N2 - Objective Chromosomal gains at 3q26, 5p15 and 20q13 have been described in cervical precancer and cancer. We evaluated a novel fluorescence in situ hybridization (FISH) assay that detects gains at these three loci simultaneously as a possible biomarker for detecting cervical precancer. Methods Chromosomal copy numbers at 3q26, 5p15, 20q13 and the centromere of chromosome7 (cen7) in liquid-based cytology specimens from 168 women enrolled in the Biopsy Study were determined by FISH. The number of cells with ≥ 3 or ≥ 4 signals for a genomic locus was enumerated and diagnostic test performance measures were ...

Three-dimensional structured illumination microscopy (3D-SIM) has opened up new possibilities to study nuclear architecture at the ultrastructural level down to the ~100 nm range. We present first results and assess the potential using 3D-SIM in combination with 3D fluorescence in situ hybridization (3D-FISH) for the topographical analysis of defined nuclear targets. Our study also deals with the concern that artifacts produced by FISH may counteract the gain in resolution. We address the topography of DAPI-stained DNA in nuclei before and after 3D-FISH, nuclear pores and the lamina, chromosome territories, chromatin domains, and individual gene loci. We also look at the replication patterns of chromocenters and the topographical relationship of Xist-RNA within the inactive X-territory. These examples demonstrate that an appropriately adapted 3D-FISH/3D-SIM approach preserves key characteristics of the nuclear ultrastructure and that the gain in information obtained by 3D-SIM yields new insights into

TY - JOUR. T1 - Detection of p16, RB, CDK4, and p53 gene deletion and amplification by fluorescence in situ hybridization in 96 gliomas. AU - Perry, Arie. AU - Anderl, Kari. AU - Borell, Tom J.. AU - Kimmel, David W.. AU - Wang, Chiao H.. AU - OFallon, Judith R.. AU - Feuerstein, Burt G.. AU - Scheithauer, Bernd W.. AU - Jenkins, Robert Brian. PY - 1999. Y1 - 1999. N2 - Inactivation of the p53 gene is a common early event of astrocytoma tumorigenesis. Alternatively, since the p16, retinoblastoma (RB), and CDK4 genes have been implicated in malignant progression, detection of losses or amplifications of these genes in gliomas could be diagnostically, prognostically, and therapeutically important. We obtained smear preparations from 96 diffuse gliomas and 10 nonneoplastic specimens. Dual-color fluorescence in situ hybridizations using paired probes for CEN9/p16, CEN8/RB, CEN17/p53, and CEN12/CDK4 were performed and revealed expected frequencies of abnormalities, except for p53 losses, which were ...

Small RNA Detection by in Situ Hybridization Methods. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.

Hardy, R R.; Hayakawa, K; Haaijman, J; and Herzenberg, L A., B lymphocyte subpopulations identifiable by two color fluorescence analysis. Abstr. (1982). Subject Strain Bibliography 1982. 1833 ...

A molecular cytogenetic map of sorghum chromosome 1: Fluorescence in situ hybridization analysis with mapped bacterial artificial chromosomes - Texas A&M University (TAMU) Scholar profile, educations, publications, research, recent courses, and student works

Segmental duplicons (SDs) predispose to an increased frequency of chromosomal rearrangements. These rearrangements can cause a diverse range of phenotypes due to haploinsufficiency, in cis positional effects or gene interruption. Genomic microarray analysis has revealed gene dosage changes adjacent to duplicons, but the high degree of similarity between duplicon sequences has confounded unequivocal assignment of chromosome breakpoints within these intervals. In this study, we localize rearrangements within duplicon-enriched regions of Angelman/Prader-Willi (AS/PWS) syndrome chromosomal deletions with fluorescence in situ hybridization (FISH). Breakage intervals in AS deletions were localized recursively with short, coordinate-defined, single copy (SC) and low copy (LC) genomic FISH probes. These probes were initially coincident with duplicons and regions of previously reported breakage in AS/PWS. Subsequently, probes developed from adjacent genomic intervals more precisely delineated deletion breakage

FISH-positive criteria have been used according to the corresponding genes. FGFR1 high-level amplification is defined as copy number ≥ 9 or (1) an FGFR1/CEP8 ratio of ≥ 2.0, (2) the average number of FGFR1 signals per tumor cell nucleus ≥ 6, and (3) the percentage of tumor cells containing ≥ 15 FGFR1 signals or large clusters ≥ 10% [74,87]. According to previously published criteria, the EGFR gene copy number was classified into six FISH strata: disomy (two or fewer copies in more than 90% of cells), low trisomy (two or fewer copies in 40% or more of the cells, three copies in 10%-40% of cells, and four or more copies in less than 10% of cells), high trisomy (two or fewer copies in ≥ 40% of cells, three copies in ≥ 40% of cells, and four or more copies in less than 10% of cells), low polysomy (four or more copies in 10%-40% of cells), high polysomy (four or more copies in 40% of the cells or more), and gene amplification (defined by the presence of tight EGFR clusters and a ratio ...

A 10-year-old girl with Ullrich-Turner syndrome was found to have the novel karyotype 45,X/46,X,r(X)(p11q11)/46,X,dic(X)(p11). Fluorescence in situ hybridization (FISH) with the alpha satellite X centromere probe established the origin of the small ring chromosome. Scanning a large number of cells by interphase FISH showed that the dicentric (X) was the least prevalent cell line. The common breakpoint of Xp11 suggests a sequence of errors as the mechanism whereby these 3 distinct cell lines have arisen.

TY - JOUR. T1 - High rate of neoplastic cells with genetic abnormalities in proliferation centers of chronic lymphocytic leukemia. AU - Balogh, Zsófia. AU - Reiniger, Lilla. AU - Rajnai, Hajnalka. AU - Csomor, Judit. AU - Szepesi, Ágota. AU - Balogh, Anikó. AU - Deák, Linda. AU - Gagyi, Éva. AU - Bödör, Csaba. AU - Matolcsy, A.. PY - 2011/6. Y1 - 2011/6. N2 - In lymph nodes, chronic lymphocytic leukemia (CLL) cells (prolymphocytes and paraimmunoblasts) form proliferation centers (PCs), which are also known as pseudofollicles. To reveal whether PCs play a role in the accumulation of genetic alterations in CLL, we compared deletion at 11q22.3, 13q14.3, and 17p13.1 loci and trisomy 12 by the fluorescence in situ hybridization (FISH) technique in PCs versus surrounding small lymphocytes (SLs) in 12 formalin-fixed paraffin-embedded (FFPE) lymph nodes. The FFPE sections were stained with methylene blue and PCs were marked by laser beam. Subsequent FISH analysis was performed, relocalizing the ...

Rosetta Genomics Ltd. operates as a genomic diagnostics company worldwide. The company ?s microRNA technologies based diagnostic tests include RosettaGX Cancer Origin for the identification of the primary site of metastatic cancer; mi-KIDNEY, a kidney tumor classification test for pathology samples; RosettaGX Reveal for the diagnosis of indeterminate thyroid fine-needle aspirate samples; and mi-LUNG diagnostic tests. It also provides UroVysion, a urine-based Fluorescence In Situ Hybridization (FISH) assay that is intended for use in conjunction with and not in lieu of current standard diagnostic procedures, as an aid for initial diagnosis of bladder carcinoma in patients with hematuria and subsequent monitoring for tumor recurrence in patients previously diagnosed with bladder cancer; and ERG/PTEN, which are FISH-based prognostic tests in prostate cancer. In addition, the company offers ALK/ROS1 that are FISH-based predictive tests indicated for patients who are diagnosed with late stage lung ...

XMP are chromosome-specific and comprise mouse whole chromosome painting probes which are directly labeled with an emitting fluorochrome.

In situ hybridization allows us to learn more about the geographical location of, for example, the messenger RNA (mRNA) in a cell or tissue. It can also tell us where a gene is located on a chromosome. Obviously, a detection system must be built into the technique to allow the cytochemist to visualize and map the geography of these molecules in the cells in question.. When in situ hybridization was first introduced, it was applied to isolated cell nuclei to detect specific DNA sequences. Early users applied the techniques to isolated chromosomal preparations in order to map the location of genes in those chromosomes. The technique has also been used to detect viral DNA in an infected cell. In situ hybridization of RNA has also been used to show that RNA synthesis (transcription) occurs in the nucleus, while protein synthesis (translation) occurs in the cytoplasm.. ...

Hello Debbie, As usual, I can not offer an unbiased opinion. However, InnoGenex does carry many FISH detection systems and two Her-2 probes[one DNA probe & one mRNA probe]. The systems are sensitive, fast and very consistent ; the ISH kits are currently used in diagnostic assays at major hospitals/research institutions(want a reference?). I have enclosed a manual from one of the FISH kits. If you would like the data sheets for the two Her-2 probes, let me know. Regards, Matthew Ogdie InnoGenex (925)543-1414 http://www.innogenex.com -----Original Message----- From: Jennings-Siena, Debbie [mailto:[email protected]] Sent: Tuesday, June 06, 2000 10:21 AM To: [email protected] Subject: Her 2 by FISH If your lab is doing Her 2 by Fluorescent In-situ Hybridization techniques (FISH) could you please tell me which manufacturers kit that you are using and how you like it? My pathologist would like to know whose kit has the highest reputation, I would also like to know about ...

Despite significant advances in molecular genetic approaches, fluorescence in situ hybridization (FISH) remains the gold standard for the diagnostic evaluation of genomic aberrations in patients with chronic lymphocytic leukemia (CLL). Efforts to improve the diagnostic utility of molecular cytogenetic testing have led to the expansion of the traditional 4-probe CLL FISH panel. Not only do these efforts increase the cost of testing, they remain hindered by the inherent limitations of FISH studies - namely the inability to evaluate genomic changes outside of the targeted loci. While array-based profiling and next generation sequencing (NGS) have critically expanded our understanding of the molecular pathogenesis of CLL, these methodologies are not routinely used by diagnostic laboratories to evaluate copy number changes or the mutational profile of this disease. Mitogenic stimulation of CLL specimens with CpG-oligonucleotide (CpG-ODN) has been identified as a reliable and reproducible means of ...

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Fig. 2. Standard diagnostic techniques are not optimal for the routine detection of ALK-rearranged NSCLC. Representative ALK-rearranged (A-F) and ALK germ-line (G-I) tumors analyzed by FISH using probes flanking the ALK gene (A, D, and G), standard immunohistochemical staining for ALK protein (B, E, and H), and tyramide-amplified immunohistochemical staining for ALK protein (C, F, and I). Red arrows, split FISH probes characteristic of an ALK rearrangement; yellow arrow, nonsplit FISH probes characteristic of ALK germ-line. ...

An ultrasonic probe having an electronic radial transducer is disclosed. In the ultrasonic probe, a plurality of piezoelectric elements having electrodes on both surfaces thereof are arranged and juxtaposed on a circular backing member. A doughnut shaped substrate is provided on an end surface of the piezoelectric elements of the electronic radial transducer and having connection points arranged in a signal pattern conductive to one each electrode of the piezoelectric elements. Additionally, at least one overlapping substrate having connection points arranged in accordance with the respective connection points of the signal pattern of the doughnut shaped substrate.

An analysis of the chromosomal aberrations and DNA ploidy in the interphase nuclei of seven human osteosacomas was preformed by double-target fluorescence in situ hybridization (FISH) and DNA cytofluorometry. The FISH study of the numerical aberrations in chromosomes 1 and 17 or the structural aberrations in chromosome arm 1p or 17p was carried out by using four locus specific DNA markers, with one pair consisting of 1q12 and 1p36 and the other pair consisting of the 17 cemtromere and 17p13.3. There was no significant differences in the percentage of deletions in chromosome 1 and 17 between osteosarcomas and normal tissues ...

In situ hybridization is a widely employed technique allowing spatial visualization of gene expression in fixed specimens. It has greatly advanced our understanding of biological processes, including developmental regulation. In situ protocols are today routinely followed in numerous laboratories, a …

Ovrednotili so zmogljivost štiribarvnega ERG/PTEN QD ISH testa pri optimalni pred obdelavi vzorcev. Signale so prešteli pri 389 vzorcih, ki so jih pridobili iz desetih tkiv prostate. Izkazalo se je, da je bilo 91% obarvanj sprejemljivih tako za ERG3p ter ERG5p, kot tudi za PTEN ter CEN10. Ne sprejemljivih je bilo samo 36 od 386 preparatov, od katerih je pri 28 prišlo do visokega ozadja pri QD655, pri osemih pa so bili signali prešibki oziroma jih sploh ni bilo. Med preparati s sprejemljivim obarvanjem je bilo 280 takih, ki so jih pridobili iz osem vzorcev benignega tkiva prostate in 70 takih, ki so bili pridobljeni iz dveh vzorcev tkiv z rakom prostate. Naključno so izbrali 19 preparatov pripravljenih iz dveh benignih tkiv in dveh tkiv z rakom prostate, katerim so prešteli signale za ERG3p, ERG5p, PTEN in CEN10 na jedro celice v treh različnih dneh. Tako pridobljeni podatki so pokazali, da so rezultati obarvanja konstanti skozi vse dni štetja. Pri preparatih iz vzorca benignega tkiva se ...

Sigma-Aldrich offers abstracts and full-text articles by [Darcy A Kerr, Kshitij S Arora, Krishnan K Mahadevan, Jason L Hornick, Jeffrey F Krane, Miguel N Rivera, David T Ting, Vikram Deshpande, William C Faquin].

Fluorescent In situ Hybridization, is fast and specific technique which can be used to detect alteration in numerical and structural chromosomal anomalies. With technological advancement multicolour FISH (m-FISH or SKY FISH) can be carried out and single probe (for just one chromosome) can also be carried out as per the need of case. It can also be employed in small deletion, duplication, translocation and chromatin fusion attributed disorders. Certain key features that make FISH a valuable tool is turn around time, which is less than 24 hours. Another advantage of FISH based analysis is it can be carried out in interphase nuclei, ruling out the need of culture and thus playing a crucial role in non dividing cells or dead cells where chromosomal analysis is not possible. Few of FISH based test are enlisted below and situations where those tests can be utilized.. ...

Weve found through our studies that fish do have a memory. … Its the same way for the fishs buddies that observed that fish being caught, too. When they see the lure come past, they are going to remember and they are going to avoid it. The same holds true for lakes that are exposed to heavy fishing pressure ...

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The principal findings of this study are the following: (a) the results of FISH/CISH analysis done on small-sized specimens represent a successful method for establishing the EGFR/HER2 gene content in NSCLC; (b) as reported in the literature, gain of EGFR and HER2 genes is more frequently a consequence of Chr7 and Chr17 polysomy, respectively, rather than gene amplification; and (c) concurrent polysomy and, less specifically, concurrent trisomy of EGFR and HER2 may be considered as positive markers for selecting NSCLC patients eligible for TKI treatment.. The first aim of our study was to validate the EGFR/HER2 gene study by FISH or CISH in very small tissue samples of lung cancers. In our series, in 68% of cases, the first diagnosis of NSCLC was obtained from small biopsies or cytologic samples. This limited the possibility of characterizing the histotype of tumors in 7% of cases. Several studies have reported successful use of FISH and CISH in alcohol-fixed fine-needle aspiration cytology or ...

The MYH11/CCP16 FISH Probe Kit is designed to detect the human MYH11 gene located on chromosome band 16p13.11 along with the number of chromosome 16 copies per

Hello Everyone, I was hoping to get some insight on the results of a FISH test. a quick background - gross hematuria 3 months ago, MRI negative showed cyst on...

PURPOSE: HER2 is a clinically important tumor marker in breast cancer; however, there is controversy regarding which method reliably measures HER2 status. We compared three HER2 laboratory methods: immunohistochemistry (IHC), fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR), to predict disease-free survival (DFS) and overall survival (OS) after adjuvant doxorubicin-based therapy in node-positive breast cancer patients. METHODS: This is a Cancer and Leukemia Group B (CALGB) study, using 524 tumor blocks collected from breast cancer patients registered to clinical trial CALGB 8541. IHC employed CB11 and AO-11-854 monoclonal antibodies; FISH used PathVysion HER2 DNA Probe kit; PCR utilized differential PCR (D-PCR) methodology. RESULTS: Cases HER2 positive by IHC, FISH and D-PCR were 24%, 17%, and 18%, respectively. FISH and IHC were clearly related (kappa = 64.8%). All three methods demonstrated a similar relationship for DFS and OS. By any method, for patients with HER2

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Please Note: ProSci now has over 700 different lots available for a wide range of tissues. Many of these lots are not listed online. If you dont see what you are looking for please call us.Formalin-fixed, paraffin-embedded tissue is useful for a variety of applications including the targeting of proteins, RNA, and DNA by means of Immunohistochemistry (IHC), in-situ hybridization (ISH) and fluorescence in-situ hybridization (FISH), respectively. Some tissue types may require extra attention as they may contain high levels of endogenous biotin, peroxidase, or autofluorescence ...

Single-molecule RNA FISH was conducted as described previously (Lee et al., 2013a). Cells grown overnight in AFM were fixed with formaldehyde (final concentration 3.7% vol/vol) at 30°C for 1 h on the shaker and washed twice with ice-cold buffer B (1.2 M sorbitol and 0.1 M potassium phosphate, pH 7.5). The fixed cells were resuspended in 1 ml spheroplasting buffer (10 ml buffer B and 2 mM vanadyl ribonucleoside complex) and transferred to a new RNase-free microcentrifuge tube. 1.5 mg Zymolase (MP Biomedicals) was added to the cells and incubated at 37°C for 35 min for wild-type and 10 min for whi3Δ cells, and cells were washed twice with buffer B. Cells were resuspended in 1 ml RNase-free 70% overnight at 4°C. RNA FISH probes (Biosearch Technologies) were resuspended in 20 µl TE buffer (10 mM TrisCl and 1 mM EDTA, pH 8.0). Then probes were diluted 1:10 from initial probe stock (250 µM in TE buffer). On the next day, cells were washed with wash buffer (20× SSC, 10% vol/vol deionized ...

Visualize Gene Expression & Genetic Variations in Tissues:. Applications of RNAscope® and BaseScopeTM ISH technology. The nervous system consists of numerous specialized cell types that remain to fully cataloged and characterized at the molecular level. Due to the high degree of structural and functional heterogeneity and the intricate spatial organization of these cells, it is of special importance to analyze gene expression in the presence of full morphological and spatial contexts. Due to the lack of specific antibody reagents, especially for lncRNAs, G-protein coupled receptors (GPCRs), and ion channels, mapping of specific transcripts by in situ hybridization offers an excellent alternative approach. The RNAscope® assay provides a powerful method to detect gene expression within the spatial and morphological tissue context. BaseScopeTM is a novel in situ hybridization technology that allows visualization of splice junctions between adjacent exons and/or retained introns in highly specific ...