Current techniques for single-cell lysis

Brown, Robert B.; Audet, Julie

doi:10.1098/rsif.2008.0009.focus

Cited by 228 publications

(209 citation statements)

References 35 publications

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“…Strong ionic detergents such as SDS are one of the most widely used cell-lysis techniques for biochemical assays, since they provide fast cell lysis on the order of seconds without the need for specialized equipment. 37 For optimization, we analyzed the impact of combining the hybridization buffer with detergent, protease, and RNase inhibitor. In all assays, we analyzed the endogenous miR-21 signals from 5000 3T3 cells.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

Encoded Hydrogel Microparticles for Sensitive and Multiplex microRNA Detection Directly from Raw Cell Lysates

et al. 2016

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In recent years, microRNAs (miRNAs) have emerged as promising diagnostic markers because of their unique dysregulation patterns under various disease conditions and high stability in biological fluids. However, current methods of analyzing miRNA levels typically require RNA isolation, which is cumbersome and time-consuming. To achieve high-throughput and accurate miRNA profiling, this study eliminates the need for purification steps by detecting miRNA directly from raw cellular lysate using nonfouling polyethylene glycol microparticles. In contrast to recent studies on direct miRNA measurements from cell lysate, our hydrogel-based system provides high-confidence quantification with robust performance. The lysis buffer for the assay was optimized to maximize reaction and labeling efficiency, and this assay has a low limit of detection (<1000 cells) without target amplification. Additionally, the capability for multiplexing was demonstrated through analyzing the levels of three endogenous miRNAs in 3T3 cell lysate. This versatile platform holds great potential for rapid and reliable direct miRNA quantification in complex media, and can be further extended to single-cell analysis by exploiting the flexibility and scalability of our system.

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Section: ■ Experimental Sectionmentioning

confidence: 99%

Encoded Hydrogel Microparticles for Sensitive and Multiplex microRNA Detection Directly from Raw Cell Lysates

et al. 2016

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“…In a different study, SDS has been used for lysing cells to recover intracellular components (Brown and Audet, 2008).…”

Section: Introductionmentioning

confidence: 99%

Surfactant-assisted direct biodiesel production from wet Nannochloropsis occulata by in situ transesterification/reactive extraction

2016

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Please cite this article as: Salam K.A., Velasquez-Orta S.B., Harvey A.P. Effect Surfactant-assisted direct biodiesel production from wet Nannochloropsis occulata by in situ transesterification/reactive extraction. Biofuel Research Journal 9 (2016) HIGHLIGHTSSurfactant assisted in situ transesterification of wet algae was studied.A surfactant catalyst ("ZDS") produced high yields in Nannochloropsis occulata.Inclusion of SDS in increased FAME production in the wet algae.The process was not adversely affected by water in the algae up to 20%. This article reports an in situ transesterification/reactive extraction of Nannochloropsis occulata for fatty acid methyl ester (FAME) production using H2SO4, sodium dodecyl sulphate (SDS) plus H2SO4 and zirconium dodecyl sulphate (ZDS). A maximum 67 % FAME yield was produced by ZDS. Effect of inclusion of sodium dodecyl sulphate (SDS) in H2SO4 for FAME enhancement and water tolerance was also studied by hydrating the algae with 10 % -30 % distilled water (w/w) dry algae. Treatment with SDS in H2SO4 increases the FAME production rate and water tolerance of the process. Inclusion of SDS in H2SO4 produced a maximum 98.3 % FAME yield at 20 % moisture in the algae. The FAME concentration began to diminish only at 30 % moisture in the algae. Furthermore, the presence of a small amount of water in the biomass or methanol increased the lipid extraction efficiency, improving the FAME yield, rather than inhibiting the reaction. ARTICLE INFO ABSTRACT

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“…High strength of electric fields, long exposure of time, or both could result in rupture of the cell membrane and cell lysis. 27,28 The field strength required for lysis of mammalian cells was in the order of magnitude of 10 6 V=m. 29 The electric field strength applied herein (approximate 5 Â 10 5 V=m, 1 MHz) was lower than the threshold strength of cell lysis to maintain the viability of cells.…”

Section: Resultsmentioning

confidence: 99%

A handheld preconcentrator for the rapid collection of cancerous cells using dielectrophoresis generated by circular microelectrodes in stepping electric fields

Jen

Chang

2011

Biomicrofluidics

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The ability to concentrate biological cells, such as circulating tumor cells, circulating fetal cells, and stem cells, is an important issue in medical diagnostics and characterization. The present study develops a handheld device capable of effectively preconcentrating cancerous cells. Circular microelectrodes were designed to generate a stepping electric field by switching the electric field to an adjacent electrode pair by relays. Cancerous cells with a positive dielectrophoretic response are guided toward the center of the circular microelectrodes due to the region of high electric field between the adjacent electrodes being gradually decreased in the direction of the stepping electric field. Numerical simulations of the electric fields were performed to demonstrate the concept of the proposed design. The preconcentration of HeLa cells, which are a human cervical carcinoma cell line, was achieved in 160 s with an efficiency of around 76%, with an applied peak-to-peak voltage of 16 V at a frequency of 1 MHz.

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Current techniques for single-cell lysis

Cited by 228 publications

References 35 publications

Encoded Hydrogel Microparticles for Sensitive and Multiplex microRNA Detection Directly from Raw Cell Lysates

Encoded Hydrogel Microparticles for Sensitive and Multiplex microRNA Detection Directly from Raw Cell Lysates

Surfactant-assisted direct biodiesel production from wet Nannochloropsis occulata by in situ transesterification/reactive extraction

A handheld preconcentrator for the rapid collection of cancerous cells using dielectrophoresis generated by circular microelectrodes in stepping electric fields

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