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Single cell western blotting

Single cell western blotting

引用

17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013

作者： Hughes, Alex J. Spelke, Dawn P. Xu, Zhuchen Schaffer, David V. Herr, Amy E. Department of Bioengineering University of California UC Berkeley Berkeley CA United States UC Berkeley-UCSF Graduate Program in Bioengineering UC Berkeley Berkeley CA United States Department of Chemical Engineering UC Berkeley Berkeley CA United States Helen Wills Neuroscience Institute UC Berkeley Berkeley CA United States California Institute for Quantitative Biosciences UC Berkeley Berkeley CA United States

Understanding protein-mediated cell signaling and differentiation processes requires capturing response heterogeneity for single cells in large populations. We report single-cell western blots on standard microscope s... 详细信息

ISBN: (纸本)9781632666246

关键词： Proteins

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TOWARDS SINGLE-CELL PROTEOFORM PROFILING: ON-CHIP ISOELECTRIC FOCUSING IN IMMOBILIZED PH GRADIENT GELS

TOWARDS SINGLE-CELL PROTEOFORM PROFILING: ON-CHIP ISOELECTRI...

引用

2022 Solid-State Sensors, Actuators and Microsystems Workshop, Hilton Head 2022

作者： Lomeli, Gabriela Herr, Amy E. UC Berkeley-UCSF Graduate Program in Bioengineering CA United States Department of Bioengineering University of California BerkeleyCA United States Chan Zuckerberg Biohub San FranciscoCA United States

ISBN: (纸本)9781940470047

Microfluidic tools are an essential capability in understanding cell-to-cell heterogeneity. Similarly, precision microanalytical tools can play an outsize role in measuring molecule-to-molecule heterogeneity, especially as protein post-translational modifications create proteoforms that execute regular and aberrant cellular functions. Although highly related, slight variations in protein charge contribute to proteoform heterogeneity. Electrophoretic methods, including immobilized pH gradient isoelectric focusing (IPG-IEF), provide the resolution to measure subtle protein-to-protein differences. However, conventional IPG-IEF can be low-throughput and requires large starting sample amounts, precluding single-cell analysis. Here, we present the first polydimethylsiloxane (PDMS)-based, immunoprobed, microfluidic IPG-IEF device to our knowledge. We demonstrate separation performance on par with our single-cell IEF device, including resolving analytes differing by only 0.1 pH. © 2022 TRF.

关键词： Electrophoresis

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FABRICATION OF STABLE GRADIENTS IN A HYDROGEL-FILLED MICROFLUIDIC DEVICE 25

FABRICATION OF STABLE GRADIENTS IN A HYDROGEL-FILLED MICROFL...

引用

25th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2021

ISBN: (纸本)9781733419031

Gradients play a key role in mimicking biological conditions in situ and in separating biological species for analysis. Current methods to generate microscale gradients typically require high reagent consumption and enclosed microfluidic devices. Here, we introduce a simple and controllable method to generate stable immobilized gradients in a hydrogel-filled PDMS channel using only 7 µL of reagent. The versatile fabrication technique forms gradients of compounds containing an abstractable hydrogen (C-H) bond. Moreover, the PDMS can be detached from the glass substrate for open access to the gradient gel. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

关键词： Hydrogels

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Disassembly of a core-satellite nanoassembled substrate for colorimetric biomolecular detection

Disassembly of a core-satellite nanoassembled substrate for ...

引用

作者： Waldeisen, John R. Wang, Tim Ross, Benjamin M. Lee, Luke P. UC Berkeley/UCSF Graduate Program in Bioengineering Biomolecular Nanotechnology Center University of California Berkeley CA 94720 United States Department of Bioengineering Biomolecular Nanotechnology Center University of California Berkeley CA 94720 United States

The disassembly of a core-satellite nanostructured substrate is presented as a colorimetric biosensor observable under dark-field illumination. The fabrication method described herein utilizes thiol-mediated adsorption and streptavidin-biotin binding to self-assemble core-satellite nanostructures with a sacrificial linking peptide. Biosensing functionality is demonstrated with the protease trypsin, and the optical properties of the nanoassemblies are characterized. A figure of merit is presented to determine the optimal core and satellite size for visual detection. Nanoassemblies with 50 nm cores and 30 or 50 nm satellites are superior as these structures achieve an orange to green color shift greater than 70 nm that is easily discernible by the naked eye. This colorimetric substrate may prove to be a favorable alternative to liquid-based colloidal sensors and a useful visual readout mechanism for point-of-care microfluidic diagnostic assays. © 2011 American Chemical Society.

关键词： Biosensors

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Information-Theoretic Design for High-Dimensional Computational Imaging

Information-Theoretic Design for High-Dimensional Computatio...

引用

Computational Optical Sensing and Imaging, COSI 2024 - Part of Optica Imaging Congress

作者： Markley, Eric Kabuli, Leyla Chien, Tiffany Pinkard, Henry Waller, Laura UCB/UCSF Joint Graduate Program in Bioengineering University of California Berkeley United States Department of Electrical Engineering and Computer Sciences University of California Berkeley United States

We present a technique for information-theoretic optimization of computational imaging systems demonstrated in snapshot 3D microscopy. By directly evaluating measurement quality and decoupling optimization from downst... 详细信息

关键词： Decoding

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Spectral DefocusCam: hyperspectral imaging using defocus and a spectral filter array

Spectral DefocusCam: hyperspectral imaging using defocus and...

引用

Computational Optical Sensing and Imaging, COSI 2022

作者： Foley, Christian Monakhova, Kristina Yanny, Kyrollos Waller, Laura Department of Electrical Engineering and Computer Sciences University of California Berkeley United States UCB/UCSF Joint Graduate Program in Bioengineering University of California Berkeley United States

We propose a compact hyperspectral camera based on a focus-tunable lens and spectral filter array. The camera captures then fuses multiple defocused measurements to obtain a high-resolution hyperspectral volume. ©... 详细信息

ISBN: (纸本)9781557528209

关键词： Hyperspectral imaging

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MultiWienerNet: Deep learning for fast shift-varying deconvolution

MultiWienerNet: Deep learning for fast shift-varying deconvo...

引用

Computational Optical Sensing and Imaging, COSI 2021 - Part of OSA Imaging and Applied Optics Congress 2021

作者： Shuai, Richard W. Yanny, Kyrollos Monakhova, Kristina Waller, Laura Department of Electrical Engineering and Computer Sciences University of California Berkeley United States UCB/UCSF Joint Graduate Program in Bioengineering University of California Berkeley United States

We present a deep-learning method based on Wiener filters and U-Nets that performs image reconstruction in systems with spatially-varying aberrations. We train on simulated microscopy measurements and test on experime... 详细信息

ISBN: (纸本)9781557528209

关键词： Image reconstruction

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Snapshot hyperspectral imaging using a random phase mask and spectral filter array

Snapshot hyperspectral imaging using a random phase mask and...

引用

Computational Optical Sensing and Imaging, COSI 2020 - Part of Imaging and Applied Optics Congress 2020

作者： Monakhova, Kristina Yanny, Kyrollos Waller, Laura Department of Electrical Engineering and Computer Sciences University of California Berkeley United States UCB/UCSF Joint Graduate Program in Bioengineering University of California Berkeley United States

We introduce a snapshot hyperspectral imager that uses a random phase mask, repeated spectral filter array, and compressive recovery to achieve high spatial and spectral resolution in a small form factor. © 2020 ... 详细信息

ISBN: (纸本)9781557528209

关键词： Hyperspectral imaging

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SAME-CELL, SINGLE-CELL DETECTION OF PROTEIN ISOFORMS AND NucLEIC ACIDS 25

SAME-CELL, SINGLE-CELL DETECTION OF PROTEIN ISOFORMS AND NUC...

引用

25th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2021

作者： Gomez Martinez, Ana E. Rosas-Canyelles, Elisabet Modzelewski, Andrew J. Geldert, Alisha Gopal, Anjali He, Lin Herr, Amy E. Department of Bioengineering University of California Berkeley BerkeleyCA United States UC Berkeley-UCSF Graduate Program in Bioengineering United States Division of Cellular and Developmental Biology Department of Molecular & Cell Biology University of California BerkeleyCA United States

ISBN: (纸本)9781733419031

Single-cell 'omics is advancing rapidly, but there are gaps in linking nucleic acids to proteins in their myriad forms. To surmount this gap, we describe a microfluidic device that reports information on nucleic acids and proteoforms, or proteins that arise from a variety of modifications to nucleic acids or proteins, which are detected by immunoblotting (triBlot). The triBlot fractionates nuclei from the cytoplasm, allowing analysis of cytoplasmic proteoforms. Then, nuclei-laden regions are laser excised to perform PCR/RT-qPCR. We correlate DNA, mRNA, and protein expression from the same few cells, a step forward in 'omics that measure diverse protein forms. © 2021 MicroTAS 2021 - 25th International Conference on Miniaturized Systems for Chemistry and Life Sciences. All rights reserved.

关键词： Proteins

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Integrated capture, concentration, PCR, and capillary electrophoretic analysis for detection of pathogens on a chip

Integrated capture, concentration, PCR, and capillary electr...

引用

13th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2009

作者： Seo, Tae Seok Beyor, Nathaniel Yi, Lina Mathies, Richard A. Department of Chemical and Biomolecular Engineering KAIST Daejeon Korea Republic of UCSF/UC Berkeley Joint Graduate Group in Bioengineering Department of Chemistry University of California Berkeley CA 94720 United States

ISBN: (纸本)9780979806421

Here we present a fully integrated lab-on-a-chip (LOC) system that incorporates cell preconcentration, polymerase chain reaction (PCR), and capillary electrophoretic (CE) microchannel for rapid and highly sensitive pathogen detection. Using the optimized bifurcated microfludic channel design and pumping flutter scheme, the E coli O157 cells were captured on the immunomagnetic beads efficiently, and the target gene was amplified in 100 nL PCR reactor, separated in 5-cm long CE channel, and detected with a detection limit of 0.2 cfu/μL in 73 min. Even in a 1000-fold excess background of E. coli K12 cells, the 100 cfu E. coli O157 was successfully identified, demonstrating high sensitivity and capability of real-world sample analysis. © 2009 CBMS.

关键词： Capillary electrophoresis

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