Quantitative Ultrasensitive Bright-Field RNA In Situ Hybridization with RNAscope

2014 ◽  
pp. 201-212 ◽  
Author(s):  
Hongwei Wang ◽  
Nan Su ◽  
Li-Chong Wang ◽  
Xingyong Wu ◽  
Son Bui ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Bright Field ◽  
Rna In Situ Hybridization

Dual-Color Ultrasensitive Bright-Field RNA In Situ Hybridization with RNAscope

2014 ◽  
pp. 139-149 ◽  
Author(s):  
Hongwei Wang ◽  
Nan Su ◽  
Li-Chong Wang ◽  
Xingyong Wu ◽  
Son Bui ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Bright Field ◽  
Dual Color ◽  
Rna In Situ Hybridization

scholarly journals RNA In Situ Hybridization for Detecting Gene Expression Patterns in the Abdomens and Wings of Drosophila Species

2021 ◽  
Vol 4 (1) ◽  
pp. 20
Author(s):  
Mujeeb Shittu ◽  
Tessa Steenwinkel ◽  
William Dion ◽  
Nathan Ostlund ◽  
Komal Raja ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Expression Patterns ◽  
Drosophila Species ◽  
Gene Expression Patterns ◽  
Probe Design ◽  
Tissue Preparation ◽  
Design And Synthesis ◽  
Rna In Situ Hybridization

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

HPV RNA In Situ Hybridization can Inform Cervical Cytology-Histology Correlation

2017 ◽  
Vol 6 (5) ◽  
pp. S29 ◽  
Author(s):  
Joseph Coppock ◽  
Brian Willis ◽  
Mark Stoler ◽  
Anne Mills
Keyword(s):  
In Situ Hybridization ◽  
Cervical Cytology ◽  
Rna In Situ Hybridization

Cell-specific metallothionein gene expression in mouse decidua and placentae

Development ◽  
1989 ◽  
Vol 107 (3) ◽  
pp. 611-621 ◽  
Author(s):  
S.K. De ◽  
M.T. McMaster ◽  
S.K. Dey ◽  
G.K. Andrews
Keyword(s):  
Gene Expression ◽  
In Situ Hybridization ◽  
Embryo Implantation ◽  
Normal Pregnancy ◽  
Mrna Levels ◽  
Metallothionein Gene ◽  
Visceral Yolk Sac ◽  
Rna In Situ Hybridization ◽  
Low Levels

Oligodeoxyribonucleotide excess solution hybridization, Northern blot and in situ hybridization were used to analyze metallothionein gene expression in mouse decidua and placentae during gestation. Metallothionein (MT) -I and -II mRNA levels were constitutively elevated, 11- and 13-fold, respectively, relative to the adult liver, in the deciduum (D8), and decreased coordinately about 6-fold during the period of development when the deciduum is replaced by the developing placenta (D10-16). Coincident with this decline, levels of MT mRNA increased dramatically in the visceral yolk sac endoderm. In situ hybridization established that MT-I mRNA was present at low levels in the uterine luminal epithelium (D4), but was elevated at the site of embryo implantation exclusively in the primary decidual zone by D5, and then in the secondary decidual zone (D6-8). Although low levels of MT mRNA were detected in total placental RNA, in situ hybridization revealed constitutively high levels in the outer placental spongiotrophoblasts. Analysis of pulse-labeled proteins from decidua and placentae established that these tissues are active in the synthesis of MT. The constitutively high levels of MT mRNA in decidua were only slightly elevated following injection of cadmium (Cd) and/or zinc (Zn), whereas in placentae they increased several-fold. MT mRNA levels were equally high in decidua and experimentally induced deciduomata (D8) which establishes that decidual MT gene expression is not dependent on the presence of the embryo or some embryo-derived factor. Although the functional role of MT during development is speculative, these results establish the concept that, from the time of implantation to late in gestation, the mouse embryo is surrounded by cells, interposed between the maternal and embryonic environments, which actively express the MT genes. This suggests that MT plays an important role in the establishment and maintenance of normal pregnancy.

scholarly journals Quantitative Single-Cell Transcript Assessment of Biomarkers Supports Cellular Heterogeneity in the Bovine IVD

2019 ◽  
Vol 6 (2) ◽  
pp. 42 ◽  
Author(s):  
Kangning Li ◽  
Devin Kapper ◽  
Sumona Mondal ◽  
Thomas Lufkin ◽  
Petra Kraus
Keyword(s):  
In Situ Hybridization ◽  
Single Cell ◽  
Gaussian Mixture ◽  
Cellular Heterogeneity ◽  
Rna In Situ Hybridization ◽  
Ivd Degeneration

Severe and chronic low back pain is often associated with intervertebral disc (IVD) degeneration. While imposing a considerable socio-economic burden worldwide, IVD degeneration is also severely impacting on the quality of life of affected individuals. Cell-based regenerative medicine approaches have moved into clinical trials, yet IVD cell identities in the mature disc remain to be fully elucidated and tissue heterogeneity exists, requiring a better characterization of IVD cells. The bovine coccygeal IVD is an accepted research model to study IVD mechano-biology and disc homeostasis. Recently, we identified novel IVD biomarkers in the outer annulus fibrosus (AF) and nucleus pulposus (NP) of the mature bovine coccygeal IVD through RNA in situ hybridization (AP-RISH) and z-proportion test. Here we follow up on Lam1, Thy1, Gli1, Gli3, Noto, Ptprc, Scx, Sox2 and Zscan10 with fluorescent RNA in situ hybridization (FL-RISH) and confocal microscopy. This permits sub-cellular transcript localization and the addition of quantitative single-cell derived values of mRNA expression levels to our previous analysis. Lastly, we used a Gaussian mixture modeling approach for the exploratory analysis of IVD cells. This work complements our earlier cell population proportion-based study, confirms the previously proposed biomarkers and indicates even further heterogeneity of cells in the outer AF and NP of a mature IVD.

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