HistoFlex—a microfluidic device providing uniform flow conditions enabling highly sensitive, reproducible and quantitative in situ hybridizations

Lab on a Chip ◽  
2011 ◽  
Vol 11 (22) ◽  
pp. 3896 ◽  
Author(s):  
Martin Jensen Søe ◽  
Fridolin Okkels ◽  
David Sabourin ◽  
Massimo Alberti ◽  
Kim Holmstrøm ◽  
...  
Lab on a Chip ◽  
2021 ◽  
Author(s):  
Zhiqi Zhao ◽  
Qiujin Li ◽  
Linna Chen ◽  
Yu Zhao ◽  
Jixian Gong ◽  
...  

Flexible biosensors for monitoring systems have emerged as a promising portable diagnostics platform due to their potential for in situ point-of-care (POC) analytic devices. Assessment of biological analytes in sweat...


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Lenka Ulrychová ◽  
Pavel Ostašov ◽  
Marta Chanová ◽  
Michael Mareš ◽  
Martin Horn ◽  
...  

Abstract Background The blood flukes of genus Schistosoma are the causative agent of schistosomiasis, a parasitic disease that infects more than 200 million people worldwide. Proteases of schistosomes are involved in critical steps of host–parasite interactions and are promising therapeutic targets. We recently identified and characterized a group of S1 family Schistosoma mansoni serine proteases, including SmSP1 to SmSP5. Expression levels of some SmSPs in S. mansoni are low, and by standard genome sequencing technologies they are marginally detectable at the method threshold levels. Here, we report their spatial gene expression patterns in adult S. mansoni by the high-sensitivity localization assay. Methodology Highly sensitive fluorescence in situ RNA hybridization (FISH) was modified and used for the localization of mRNAs encoding individual SmSP proteases (including low-expressed SmSPs) in tissues of adult worms. High sensitivity was obtained due to specifically prepared tissue and probes in combination with the employment of a signal amplification approach. The assay method was validated by detecting the expression patterns of a set of relevant reference genes including SmCB1, SmPOP, SmTSP-2, and Sm29 with localization formerly determined by other techniques. Results FISH analysis revealed interesting expression patterns of SmSPs distributed in multiple tissues of S. mansoni adults. The expression patterns of individual SmSPs were distinct but in part overlapping and were consistent with existing transcriptome sequencing data. The exception were genes with significantly low expression, which were also localized in tissues where they had not previously been detected by RNA sequencing methods. In general, SmSPs were found in various tissues including reproductive organs, parenchymal cells, esophagus, and the tegumental surface. Conclusions The FISH-based assay provided spatial information about the expression of five SmSPs in adult S. mansoni females and males. This highly sensitive method allowed visualization of low-abundantly expressed genes that are below the detection limits of standard in situ hybridization or by RNA sequencing. Thus, this technical approach turned out to be suitable for sensitive localization studies and may also be applicable for other trematodes. The results suggest that SmSPs may play roles in diverse processes of the parasite. Certain SmSPs expressed at the surface may be involved in host–parasite interactions. Graphic abstract


2012 ◽  
Vol 7 (4) ◽  
pp. 045007 ◽  
Author(s):  
Sanjukta Guha Thakurta ◽  
Robert Miller ◽  
Anuradha Subramanian

Genetics ◽  
1996 ◽  
Vol 143 (1) ◽  
pp. 365-374 ◽  
Author(s):  
Allan R Lohe ◽  
Daniel L Hartl

Abstract An important goal in molecular genetics has been to identify a transposable element that might serve as an efficient transformation vector in diverse species of insects. The transposable element mariner occurs naturally in a wide variety of insects. Although virtually all mariner elements are nonfunctional, the Mosl element isolated from Drosophila mauritiana is functional. Mosl was injected into the pole-cell region of embryos of D. virilis, which last shared a common ancestor with D. mauritiana 40 million years ago. Mosl PCR fragments were detected in several pools of DNA from progeny of injected animals, and backcross lines were established. Because Go lines were pooled, possibly only one transformation event was actually obtained, yielding a minimum frequency of 4%. Mosl segregated in a Mendelian fashion, demonstrating chromosomal integration. The copy number increased by spontaneous mobilization. In situ hybridization confirmed multiple polymorphic locations of Mosl. Integration results in a characteristic 2-bp TA duplication. One Mosl element integrated into a tandem array of 370-bp repeats. Some copies may have integrated into heterochromatin, as evidenced by their ability to support PCR amplification despite absence of a signal in Southern and in situ hybridizations.


RSC Advances ◽  
2016 ◽  
Vol 6 (87) ◽  
pp. 84200-84208 ◽  
Author(s):  
Adil Sultan ◽  
Sharique Ahmad ◽  
Faiz Mohammad

We report the synthesis of polypyrrole (PPy) and polypyrrole/silicon carbide nanocomposites (PPy/SiC) and PPy/SiC/dodecylbenzenesulfonic acid (DBSA) by in situ chemical polymerization and their application as sensors for the detection of highly toxic chlorine gas.


Sign in / Sign up

Export Citation Format

Share Document