A microfluidic device for TEM sample preparation

Hauser, Janosch; Kylberg, Gustaf; Colomb-Delsuc, Mathieu; Stemme, Göran; Sintorn, Ida‐Maria; Roxhed, Niclas

doi:10.1039/d0lc00724b

Cited by 13 publications

(6 citation statements)

References 20 publications

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“…The device was fabricated using lamination technology, as described previously. ,, Individual layers of hydrophilic sheets, adhesive tapes, magnetic tapes, and paper were structured using a laser cutter (VLS 2.30, Universal Laser Systems, Austria). Blood filters were cut into 10 × 10 mm 2 pieces using a scalpel.…”

Section: Methodsmentioning

confidence: 99%

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

et al. 2022

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Patient-centric sampling strategies, where the patient performs self-sampling and ships the sample to a centralized laboratory for readout, are on the verge of widespread adaptation. However, the key to a successful patient-centric workflow is user-friendliness, with few noncritical user interactions, and simple, ideally biohazard-free shipment. Here, we present a capillary-driven microfluidic device designed to perform the critical biomarker capturing step of a multiplexed immunoassay at the time of sample collection. On-chip sample drying enables biohazard-free shipment and allows us to make use of advanced analytics of specialized laboratories that offer the needed analytical sensitivity, reliability, and affordability. Using C-Reactive Protein, MCP1, S100B, IGFBP1, and IL6 as model blood biomarkers, we demonstrate the multiplexing capability and applicability of the device to a patient-centric workflow. The presented quantification of a biomarker panel opens up new possibilities for e-doctor and e-health applications.

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Section: Methodsmentioning

confidence: 99%

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

et al. 2022

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“…Using open microfluidic platforms will reduce the dimensions and maintain the general form factor of the microwell plate, thereby minimizing surface exposure and enabling sample recovery through nano-pipetting [ 1 , 25 ]. Other approaches have been developed by contact pin-printing methods in which pipetting robots automatically dispense liquids at the Transmission Electron Microscopy grid, such as a capillary-driven microfluidic single-use device [ 26 ]. These approaches have the advantages of reducing the volumes of the sample as well as providing automation possibilities, while they also have some disadvantages, such as the need for special instrumentation, and representing a significant increase in complexity and being more time consuming than the manual protocol [ 26 ].…”

Section: Sample Preparationmentioning

confidence: 99%

“…Other approaches have been developed by contact pin-printing methods in which pipetting robots automatically dispense liquids at the Transmission Electron Microscopy grid, such as a capillary-driven microfluidic single-use device [ 26 ]. These approaches have the advantages of reducing the volumes of the sample as well as providing automation possibilities, while they also have some disadvantages, such as the need for special instrumentation, and representing a significant increase in complexity and being more time consuming than the manual protocol [ 26 ]. Mukhitov et al suggested a microfluidic device for the preparation of the grids, in which the grid is contained in a microfluidic canal and the liquid for sample preparation is driven by an external pressure pump, thereby improving preparation consistency [ 27 ].…”

Section: Sample Preparationmentioning

confidence: 99%

Single-Cell Proteomics: The Critical Role of Nanotechnology

Arias-Hidalgo

Juanes-Velasco

Landeira-Viñuela

et al. 2022

IJMS

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In single-cell analysis, biological variability can be attributed to individual cells, their specific state, and the ability to respond to external stimuli, which are determined by protein abundance and their relative alterations. Mass spectrometry (MS)-based proteomics (e.g., SCoPE-MS and SCoPE2) can be used as a non-targeted method to detect molecules across hundreds of individual cells. To achieve high-throughput investigation, novel approaches in Single-Cell Proteomics (SCP) are needed to identify and quantify proteins as accurately as possible. Controlling sample preparation prior to LC-MS analysis is critical, as it influences sensitivity, robustness, and reproducibility. Several nanotechnological approaches have been developed for the removal of cellular debris, salts, and detergents, and to facilitate systematic sample processing at the nano- and microfluidic scale. In addition, nanotechnology has enabled high-throughput proteomics analysis, which have required the improvement of software tools, such as DART-ID or DO-MS, which are also fundamental for addressing key biological questions. Single-cell proteomics has many applications in nanomedicine and biomedical research, including advanced cancer immunotherapies or biomarker characterization, among others; and novel methods allow the quantification of more than a thousand proteins while analyzing hundreds of single cells.

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“…Optimization of specimen preparation using non-radioactive stains Preparation of efficient embedding and even specimen spreading on the grid support is a prerequisite for an unbiased TEM assessment and imaging (Figure 1A) (Hauser et al, 2020). The stain helps to maintain the specimen integrity while ensuring a good contrast (Figure 1B) for biological samples.…”

Section: Tem Workflow For the Analysis Of Vlpsmentioning

confidence: 99%

Quality assessment of virus-like particle: A new transmission electron microscopy approach

Magalhães

Santis²,

Hussein-Gore³

et al. 2022

Front. Mol. Biosci.

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Transmission electron microscopy (TEM) is a gold standard analytical method for nanoparticle characterization and is playing a valuable role in virus-like particle (VLP) characterization extending to other biological entities such as viral vectors. A dedicated TEM facility is a challenge to both small and medium-sized enterprises (SMEs) and companies operating in low-and-middle income countries (LMICs) due to high start-up and running costs. A low-voltage TEM solution with assisted image acquisition and analysis such as the MiniTEM system, coupled with Vironova Imaging and Analysis Software (VIAS) could provide an affordable and practical alternative. The MiniTEM system has a small footprint and software that enables semi-automated data collection and image analysis workflows using built-in deep learning methods (convolutional neural networks) for automation in analysis, increasing speed of information processing and enabling scaling to larger datasets. In this perspective we outline the potential and challenges in the use of TEM as mainstream analytical tool in manufacturing settings. We highlight the rationale and preliminary findings from our proof-of-concept study aiming to develop a method to assess critical quality attributes (CQAs) of VLPs and facilitate adoption of TEM in manufacturing settings. In our study we explored all the steps, from sample preparation to data collection and analysis using synthetic VLPs as model systems. The applicability of the method in product development was verified at pilot-scale during the technology transfer of dengue VLPs development from a university setting to an LMIC- based vaccine manufacturing company, demonstrating the applicability of this analytical technique to VLP vaccine characterization.

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A microfluidic device for TEM sample preparation

Abstract: We present a capillary-driven microfluidic single-use device that prepares a TEM grid with minimal user interaction. The user only initiates the sample preparation process, waits for about one minute and then collects the TEM grid, ready for imaging.

Cited by 13 publications

References 20 publications

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

Microfluidic Device for Patient-Centric Multiplexed Assays with Readout in Centralized Laboratories

Single-Cell Proteomics: The Critical Role of Nanotechnology

Quality assessment of virus-like particle: A new transmission electron microscopy approach

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