Dataset Information

A single design strategy for dual sensitive pH probe with a suitable range to map pH in living cells.

ABSTRACT: Due to the lack of a proper imaging approach, a veracious pH map of normal and abnormal cell is still rare. In this work, we presented a rhodamine-salicylaldehyde combination (Rh-SA2) as a novel pH probe, which has dual sensitive units for both acidic and basic environment. This dual sensitive probe acts like a chameleon in living cells and offers the doubling guarantees for endocellular pH mapping. Moreover, a quantitative measurement of cellular pH changes was allowed and the endocellular pH values under drug-associated stimuli were also investigated.

SUBMITTER: Yu KK

PROVIDER: S-EPMC4614390 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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A single design strategy for dual sensitive pH probe with a suitable range to map pH in living cells.

Yu Kang-Kang KK Hou Ji-Ting JT Li Kun K Yao Qian Q Yang Jin J Wu Ming-Yu MY Xie Yong-Mei YM Yu Xiao-Qi XQ

Scientific reports 20151021

Due to the lack of a proper imaging approach, a veracious pH map of normal and abnormal cell is still rare. In this work, we presented a rhodamine-salicylaldehyde combination (Rh-SA2) as a novel pH probe, which has dual sensitive units for both acidic and basic environment. This dual sensitive probe acts like a chameleon in living cells and offers the doubling guarantees for endocellular pH mapping. Moreover, a quantitative measurement of cellular pH changes was allowed and the endocellular pH v ...[more]

PMID: 26486180

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Project description:MicroRNAs (miRNAs) have been shown to play an important role in many different cellular, developmental, and physiological processes. Accordingly, numerous methods have been established to identify and quantify miRNAs. The shortness of miRNA sequence results in a high dynamic range of melting temperatures and, moreover, impedes a proper selection of detection probes or optimized PCR primers. While miRNA microarrays allow for massive parallel and accurate relative measurement of all known miRNAs, they have so far been less useful as an assay for absolute quantification. Here, we present a microarray based approach for global and absolute quantification of miRNAs. The method relies on an equimolar pool of about 1000 synthetic miRNAs of known concentration which is used as an universal reference and labeled and hybridized in a dual colour approach on the same array as the sample of interest. Each single miRNA is quantified with respect to the universal reference outbalancing bias related to sequence, labeling, hybridization or signal detection method. We demonstrate the accuracy of the method by various spike in experiments. Further, we quantified miRNA copy numbers in liver samples and CD34(+)CD133(-) hematopoietic stem cells. Different probe designs were investigated with respect to sensitivity and selectivity of microarray hybridization. An array with 11 different variants of oligonucleotides for miR-122a and miR-16 was produced. 5 µg of respective total RNA was fluorescently labelled by 3’ ligation. Total RNA was hybridized in a dual colour approach to microarrays versus a second labelled synthetic miRNA pool. The synthetic miRNA pool consisted of 5 fmol of each of 816 non redundant miRNAs sequences and miRControl 3 sequences. Local background was subtracted from the signal to obtain the net signal intensity and the mean of the net signal intensities of 4 corresponding spots representing the same miRNA was computed for those spots only which were unflagged (empty spots, poor spots, negative spots) and for which the fluorescent intensity of the miRNAs derived from the samples of interest was two-fold the mean background value (2bkg dataset).

Dataset Information

A single design strategy for dual sensitive pH probe with a suitable range to map pH in living cells.

Publications

A single design strategy for dual sensitive pH probe with a suitable range to map pH in living cells.

Similar Datasets

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