A Multi-Method Analysis of c-erbB2/HER2 in FFPE Tissue: Correlation of a Novel and Rapid In Situ Hybridization Procedure, HER2 CISH and IHC.

2009 ◽  
Author(s):  
D. Tacha ◽  
J. Alba Dominguez ◽  
J. Gutierrez Bernal ◽  
R. Henshall-Powell ◽  
J. Vargas ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Ffpe Tissue ◽  
Hybridization Procedure ◽  
Method Analysis

scholarly journals Ultrasensitive RNA In Situ Hybridization for Kappa and Lambda Light Chain mRNA in Marginal Zone Lymphoma Demonstrates Comparable Performance to Flow Cytometry

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S109-S109
Author(s):  
Michael Franklin ◽  
Chelsey Deel ◽  
Mohammad Vasef
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
B Cell ◽  
Light Chain ◽  
Marginal Zone ◽  
Marginal Zone Lymphoma ◽  
Ffpe Tissue ◽  
Light Chains ◽  
Light Chain Restriction

Abstract Objectives Evaluation of light chain restriction is critical to establish clonality in B-cell lymphoproliferative disorders (LPDs). Immunohistochemistry (IHC) and in situ hybridization (ISH) are commonly used to assess light chain restriction in formalin-fixed, paraffin-embedded (FFPE) tissues. However, except for cases with plasma cell differentiation, these techniques often fail to identify immunoglobulin light chains. An ultrasensitive technique, RNAscope, has been recently introduced that can identify light chains in cases of B-cell LPDs. We analyzed the utility of this ultrasensitive method in detection of clonality and correlated with flow cytometry results when available. Methods A tissue microarray was constructed using 1.6-mm diameter tissue punches of 31 FFPE tissue blocks from 27 cases that were previously characterized as marginal zone lymphoma (MZL) by a combination of morphology, IHC, and/or flow cytometry. Cases included 8 nodal and 19 extranodal MZLs. In two cases, additional blocks were included to assess reproducibility. For ultrasensitive ISH RNAscope assay, 4-µm thickness tissue sections were hybridized using kappa and lambda probes, incubated overnight, counterstained with hematoxylin, cover-slipped, and reviewed blindly without knowledge of prior flow cytometry results. Results Of 18 cases with evaluable staining, 15 were clonal and 3 were polytypic. Flow cytometry was available in 14 of these 18 cases with concordance in 13 of 14 (93%). The discordant case was polytypic by flow cytometry but kappa restricted by RNAscope. The false-negative flow results could be due to sampling issues. In six cases, staining failed and could not be evaluated. Conclusion Ultrasensitive RNAscope is a reliable assay in the detection of clonality in FFPE tissue, particularly where fresh tissue is not available for flow cytometry. In addition, our results confirm and further expand prior observations that RNAscope is a highly sensitive and specific assay with high concordance with flow cytometry.

scholarly journals Expression of MsLEC1 Transgenes in Alfalfa Plants Causes Symbiotic Abnormalities

2004 ◽  
Vol 17 (1) ◽  
pp. 16-26 ◽  
Author(s):  
Laurence M. Brill ◽  
Nancy A. Fujishige ◽  
Cheryl A. Hackworth ◽  
Ann M. Hirsch
Keyword(s):  
In Situ Hybridization ◽  
Sinorhizobium Meliloti ◽  
Northern Blot Analysis ◽  
Root Tips ◽  
Mrna Accumulation ◽  
Lectin Gene ◽  
Hybridization Procedure ◽  
Sense Orientation ◽  
Legume Lectins

Legume lectins have been proposed to have important symbiotic roles during Rhizobium-legume symbioses. To test this hypothesis, the symbiotic responses of transgenic alfalfa plants that express a portion of the putative alfalfa lectin gene MsLEC1 or MsLEC2 in either the antisense or sense orientation were analyzed following inoculation with wild-type Sinorhizobium meliloti 1021. MsLEC1-antisense (LEC1AS) plants were stunted, exhibited hypernodulation, and developed not only abnormally large nodules but also numerous small nodules, both of which senesced prematurely. MsLEC2-antisense plants were intermediate in growth and nodule number compared with LEC1AS and vector control plants. The symbiotic abnormalities of MsLEC1-sense transgene plants were similar to but milder than the responses shown by the LEC1AS plants, whereas MsLEC2-sense transgene plants exhibited symbiotic responses that were identical to those of vector and nontransgenic control plants. MsLEC1 mRNA accumulation was not detected in nodule RNA by Northern blot analysis but was localized to alfalfa nodule meristems and the adjacent cells of the invasion zone by in situ hybridization; transcripts were also detected in root meristems. A similar spatial pattern of MsLEC2 expression was found by using a whole-mount in situ hybridization procedure. Moreover, mRNAs for an orthologous lectin gene (MaLEC) were detected in white sweetclover (Melilotus alba) nodules and root tips.

Detection of hepatitis delta virus RNA by a nonradioactive in situ hybridization procedure

1992 ◽  
Vol 23 (5) ◽  
pp. 557-561 ◽  
Author(s):  
Donatella Pacchioni ◽  
Francesco Negro ◽  
Elisabetta Chiaberge ◽  
Mario Rizzetto ◽  
Ferruccio Bonino ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Hepatitis Delta Virus ◽  
Hepatitis Delta ◽  
Virus Rna ◽  
Hybridization Procedure ◽  
Nonradioactive In Situ Hybridization ◽  
Delta Virus

Localization of low abundance DNA sequences in tissue sections by in situ hybridization

1986 ◽  
Vol 81 (1) ◽  
pp. 143-162 ◽  
Author(s):  
C.W. Lo
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Detection System ◽  
Mouse Tissue ◽  
Globin Genes ◽  
Beta Globin ◽  
Tissue Sections ◽  
Specific Hybridization ◽  
Hybridization Procedure

Specific DNA sequences were loaclized in the nuclei of paraffin-embedded mouse tissue sections with in situ hybridization using a biotinylated globin probe in conjunction with a streptavidin-alkaline phosphatase detection system. Globin inserts were clearly detected in the tissue sections of transgenic mice containing 1000, 120 or 5 copies of the exogenously introduced beta-globin genes. In addition, specific hybridization signal was also obtained for the endogenous complement of beta-globin genes in the tissue sections of normal mice. These results demonstrate that this hybridization procedure is very sensitive and should be useful for characterizing the distribution of low abundance DNA sequences in cells and tissue sections.

scholarly journals Simultaneous detection of two messenger RNAs in the central nervous system: a simple two-step in situ hybridization procedure using a combination of radioactive and non-radioactive probes.

1991 ◽  
Vol 39 (11) ◽  
pp. 1575-1578 ◽  
Author(s):  
E Normand ◽  
B Bloch
Keyword(s):  
Central Nervous System ◽  
In Situ Hybridization ◽  
Simultaneous Detection ◽  
Messenger Rnas ◽  
Tissue Sections ◽  
System A ◽  
Step Procedure ◽  
Hybridization Procedure ◽  
The Central Nervous System

We present here a method enabling the simultaneous detection of two messenger RNAs in tissue sections by use of a two-step in situ hybridization procedure. Tissue sections were hybridized with a radioactive probe and coated with emulsion. The emulsion was processed for development, fixed, and a second hybridization was performed through the emulsion with a biotinylated probe subsequently revealed with streptavidin-alkaline phosphatase. This procedure allows the detection of two mRNAs without loss of signal, removal of the emulsion, or spurious reaction. The simultaneous detection of oxytocin and vasopressin mRNAs in the hypothalamus, and of dopamine receptor and neuropeptide mRNAs in the striatum, demonstrated the efficiency of the procedure. Such a two-step procedure provides a simple and flexible way to make possible comparative analysis of the localization of two mRNAs within the same tissue section.

Sign in / Sign up

Export Citation Format

Share Document