Microtiter Plate-Based Microfluidic Platforms: Sealing, Leakage Testing, and Performance of a 96-Well SPRI Device

2006 ◽  
Author(s):  
D. S. Park ◽  
M. Hupert ◽  
J. Guy ◽  
P. Datta ◽  
J.-B. Lee ◽  
...  
Keyword(s):  
High Throughput ◽  
Mold Insert ◽  
Microfluidic Devices ◽  
Microtiter Plate ◽  
Thickness Variation ◽  
Large Area ◽  
Microfluidic Platforms ◽  
Effective Manner ◽  
Micro Molding ◽  
Microtiter Plate Format

Highly parallelized biochemical analysis is a significant step toward achieving high throughput processing of patient samples for diagnosis and treatment monitoring. The standard microtiter plate is used to carry out multiple reactions for high throughput screening. By incorporating polymer microfluidic devices at each well in the microtiter plate format, the capability of the format could be significantly enhanced for high throughput processing of large numbers of biochemical samples in a cost-effective manner. Low cost replication of the microtiter plates is done using micro molding techniques, so microfabrication technology for making large area mold inserts (LAMIs) containing microfluidic devices at each well of a microtiter plate format is needed. A large area mold insert (LAMI) in the footprint of the standard microtiter plate was fabricated using an SU-8 based UV-LIGA technique. Excellent lithography results, with vertical sidewalls, were obtained by utilizing flycutting to minimize SU-8 film thickness variation and a UV filter for attenuating high absorbance UV wavelengths. Overplating of nickel in the SU-8 polymeric molds was used to make high quality metallic mold inserts with vertical sidewalls. Micro molding of polycarbonate (PC) was done using hot embossing, resulting in good replication fidelity over the large surface area. Thermal fusion bonding of the molded PC chips yielded good sealing results and the developed polymer microfluidic platforms showed good fluidic uniformity.

Rapid, high-throughput method for the quantification of thyroid hormones in rat blood serum using isotope-dilution LC–MS/MS

Bioanalysis ◽  
2020 ◽  
Vol 12 (23) ◽  
pp. 1689-1698
Author(s):  
Charles R Powley ◽  
Jinlan Dong ◽  
Bethany R Hannas ◽  
Zhihua Amanda Shen
Keyword(s):  
Thyroid Hormones ◽  
Blood Serum ◽  
High Throughput ◽  
Toxicity Testing ◽  
Microtiter Plate ◽  
Limit Of Quantification ◽  
Internal Standards ◽  
High Throughput Method ◽  
Microtiter Plate Format

Aim: Numerous guideline studies required for regulatory toxicity testing now include the measurement of the thyroid hormones 3,3′,5-triiodo-L-thyronine (T3) and L-thyroxine (T4) in blood serum from rodents. A rapid, high-throughput method for the determination of the thyroid hormones T4 and T3 is reported. Materials & methods: Sample preparation is done using a 96-well microtiter plate format. Stable isotope analogs of both hormones are used as internal standards for study and quality control samples. Results & conclusion: The validated quantification levels are T3: 10 pg/ml and T4: 1 ng/ml, with CVs of <10% at the limit of quantification and up to 50*limit of quantification. The use of isotope analog internal standards eliminates the need for quantitative transfers and complete evaporations.

scholarly journals Microfluidic based Platform for drug screening-A review

2020 ◽  
Vol 11 (3) ◽  
pp. 2999-3004
Author(s):  
Kalpana Seelam ◽  
Daisy Rani A
Keyword(s):  
Drug Screening ◽  
Dynamic Condition ◽  
Microfluidic Devices ◽  
Drug Susceptibility ◽  
Cost Models ◽  
Microfluidic Platforms ◽  
Effective Manner ◽  
2D And 3D ◽  
Body Dynamic ◽  
Very High

There is a lot of requirement to develop a preclinical rapid drug screening devices to treat the throat cancerous patients in effective manner. Very High cost models like animal based, 2d and 3d type models are static drug screening models and they are unable to mimic the human body dynamic condition. So microfluidic platform based drug screening devices will give predetermined and prominent result in rapid drug screening by mimic dynamic body conditions. Now it is required to review what are the developments from last five years in screening the drug for cancer. Recently research is going on microfluidic platforms to screen the efficacy of mixed drugs, drug tolerance, and drug susceptibility. So this study presents the review on what are the advancements in microfluidic platforms for rapid drug screening and for different cancer patients. This study also presents what are the different sensing methodologies of microfluidic devices exists to screen the drug for various cancerous tissues

scholarly journals O23: CHARACTERISING PATIENT-DERIVED COLORECTAL CANCER TISSUE-ORIGINATED ORGANOIDAL SPHEROIDS FOR HIGH-THROUGHPUT MICROFLUIDIC APPLICATIONS

2021 ◽  
Vol 108 (Supplement_1) ◽  
Author(s):  
MI Khot ◽  
M Levenstein ◽  
R Coppo ◽  
J Kondo ◽  
M Inoue ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Fluid Flow ◽  
High Throughput ◽  
Microfluidic Devices ◽  
In Vitro Models ◽  
Fluorescent Imaging ◽  
Cancer Tissue ◽  
Cell Models

Abstract Introduction Three-dimensional (3D) cell models have gained reputation as better representations of in vivo cancers as compared to monolayered cultures. Recently, patient tumour tissue-derived organoids have advanced the scope of complex in vitro models, by allowing patient-specific tumour cultures to be generated for developing new medicines and patient-tailored treatments. Integrating 3D cell and organoid culturing into microfluidics, can streamline traditional protocols and allow complex and precise high-throughput experiments to be performed with ease. Method Patient-derived colorectal cancer tissue-originated organoidal spheroids (CTOS) cultures were acquired from Kyoto University, Japan. CTOS were cultured in Matrigel and stem-cell media. CTOS were treated with 5-fluorouracil and cytotoxicity evaluated via fluorescent imaging and ATP assay. CTOS were embedded, sectioned and subjected to H&E staining and immunofluorescence for ABCG2 and Ki67 proteins. HT29 colorectal cancer spheroids were produced on microfluidic devices using cell suspensions and subjected to 5-fluorouracil treatment via fluid flow. Cytotoxicity was evaluated through fluorescent imaging and LDH assay. Result 5-fluorouracil dose-dependent reduction in cell viability was observed in CTOS cultures (p&lt;0.01). Colorectal CTOS cultures retained the histology, tissue architecture and protein expression of the colonic epithelial structure. Uniform 3D HT29 spheroids were generated in the microfluidic devices. 5-fluorouracil treatment of spheroids and cytotoxic analysis was achieved conveniently through fluid flow. Conclusion Patient-derived CTOS are better complex models of in vivo cancers than 3D cell models and can improve the clinical translation of novel treatments. Microfluidics can streamline high-throughput screening and reduce the practical difficulties of conventional organoid and 3D cell culturing. Take-home message Organoids are the most advanced in vitro models of clinical cancers. Microfluidics can streamline and improve traditional laboratory experiments.

scholarly journals Disposable Voltammetric Immunosensors Integrated with Microfluidic Platforms for Biomedical, Agricultural and Food Analyses: A Review

Sensors ◽  
2018 ◽  
Vol 18 (12) ◽  
pp. 4124 ◽  
Author(s):  
Fabiana Felix ◽  
Alexandre Baccaro ◽  
Lúcio Angnes
Keyword(s):  
Quality Management ◽  
Signal Amplification ◽  
Screen Printing ◽  
Point Of Care ◽  
Microfluidic Devices ◽  
Strong Interactions ◽  
Point Of Care Testing ◽  
Microfluidic Platforms ◽  
Broad Variety ◽  
Interesting Alternative

Disposable immunosensors are analytical devices used for the quantification of a broad variety of analytes in different areas such as clinical, environmental, agricultural and food quality management. They detect the analytes by means of the strong interactions between antibodies and antigens, which provide concentration-dependent signals. For the herein highlighted voltammetric immunosensors, the analytical measurements are due to changes in the electrical signals on the surface of the transducers. The possibility of using disposable and miniaturized immunoassays is a very interesting alternative for voltammetric analyses, mainly, when associated with screen-printing technologies (screen-printed electrodes, SPEs), and microfluidic platforms. The aim of this paper is to discuss a carefully selected literature about different examples of SPEs-based immunosensors associated with microfluidic technologies for diseases, food, agricultural and environmental analysis. Technological aspects of the development of the voltammetric immunoassays such as the signal amplification, construction of paper-based microfluidic platforms and the utilization of microfluidic devices for point-of-care testing will be presented as well.

scholarly journals Cerebrospinal Fluid Secretory Ca2+-Dependent Phospholipase A2 Activity Is Increased in Alzheimer Disease

2009 ◽  
Vol 55 (12) ◽  
pp. 2171-2179 ◽  
Author(s):  
Sonia Chalbot ◽  
Henrik Zetterberg ◽  
Kaj Blennow ◽  
Tormod Fladby ◽  
Inge Grundke-Iqbal ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Alzheimer Disease ◽  
Phospholipase A2 ◽  
Microtiter Plate ◽  
Pla2 Activity ◽  
Activity Assay ◽  
Proinflammatory Mediators ◽  
Spla2 Activity ◽  
Phospholipase A2 Activity ◽  
Microtiter Plate Format

Abstract Background: The phospholipase A2 (PLA2) family comprises multiple isoenzymes that vary in their physicochemical properties, cellular localizations, calcium sensitivities, and substrate specificities. Despite these differences, PLA2s share the ability to catalyze the synthesis of the precursors of the proinflammatory mediators. To investigate the potential of PLA2 as a biomarker in screening neuroinflammatory disorders in both clinical and research settings, we developed a PLA2 assay and determined the predominant types of PLA2 activity in cerebrospinal fluid (CSF). Methods: We used liposomes composed of a fluorescent probe (bis-Bodipy® FL C11-PC [1,2-bis-(4,4- difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-undecanoyl)-sn-glycero-3-phosphocholine]) and 1,2-dioleoyl-l-α-phosphatidylcholine as a substrate to measure CSF PLA2 activity in a 96-well microtiter plate format. We established the type of CSF PLA2 activity using type-specific inhibitors of PLA2. Results: Using 5 μL CSF per assay, our PLA2 activity assay was reproducible with CVs &lt;15% in 2 CSF samples and for recombinant secretory Ca2+-dependent PLA2 (sPLA2) in concentrations ranging from 0.25 to 1 μmol/L. This PLA2 assay allowed identification of sPLA2 activity in lumbar CSF from healthy individuals 20–77 years old that did not depend on either sex or age. Additionally, CSF sPLA2 activity was found to be increased (P = 0.0008) in patients with Alzheimer disease. Conclusions: Adult human CSF has sPLA2 activity that can be measured reliably with the assay described. This enzyme activity in the CSF is independent of both sex and age and might serve as a valuable biomarker of neuroinflammation, as we demonstrated in Alzheimer disease.

scholarly journals Harnessing intrinsic fluorescence for typing of secondary structures of DNA

2020 ◽  
Author(s):  
Michela Zuffo ◽  
Aurélie Gandolfini ◽  
Brahim Heddi ◽  
Anton Granzhan
Keyword(s):  
High Throughput ◽  
Conformational Changes ◽  
Emission Spectra ◽  
Principal Component ◽  
Secondary Structures ◽  
Intrinsic Fluorescence ◽  
Label Free ◽  
Linear Discriminant ◽  
Effective Manner

ABSTRACTDNA is polymorphic since, despite its ubiquitous presence as a double-stranded helix, it is able to fold into a plethora of other secondary structures both in vitro and in cells. Despite the considerable advances that have been made in understanding this structural diversity, its high-throughput investigation still faces severe limitations. This mainly stems from the lack of suitable label-free methods, combining a fast and cheap experimental workflow with high information content. Here, we explore the use of intrinsic fluorescence emitted by nucleic acids for this scope. After a preliminary assessment of the suitability of this phenomenon for tracking the conformational changes of DNA, we examined the intrinsic steady-state emission spectra of an 89-membered set of synthetic oligonucleotides with reported conformation (G-quadruplexes, i-motifs, single- and double stranded DNA) by means of multivariate analysis. Specifically, principal component analysis of emission spectra resulted in successful clustering of oligonucleotides into three corresponding conformational groups, albeit without discrimination between single- and double-stranded structures. Linear discriminant analysis of the same training set was exploited for the assessment of new sequences, allowing the evaluation of their G4-forming propensity. Our method does not require any labelling agent or dye, avoiding the related intrinsic bias, and can be utilized to screen novel sequences of interest in a high-throughput and cost-effective manner. In addition, we observed that left-handed (Z-) G4 structures were systematically more fluorescent than most other G4 structures, almost reaching the quantum yield of 5′-d[(G3T)3G3]-3′ (G3T), the most fluorescent G4 structure reported to date. This property is likely to arise from the similar base-stacking geometry in both types of structures.

Droplet-based microfluidics in biomedical applications

2021 ◽  
Author(s):  
Leyla Amirifar ◽  
Mohsen Besanjideh ◽  
Rohollah Nasiri ◽  
Amir Shamloo ◽  
Fatemeh Nasrollahi ◽  
...  
Keyword(s):  
Drug Delivery ◽  
High Throughput ◽  
Paradigm Shift ◽  
Cell Biology ◽  
Biomedical Applications ◽  
Molecular Detection ◽  
Droplet Microfluidics ◽  
Droplet Generation ◽  
Microfluidic Platforms ◽  
Standard Methods

Abstract Droplet-based microfluidic systems have been employed to manipulate discrete fluid volumes with immiscible phases. Creating the fluid droplets at microscale has led to a paradigm shift in mixing, sorting, encapsulation, sensing, and designing high throughput devices for biomedical applications. Droplet microfluidics has opened many opportunities in microparticle synthesis, molecular detection, diagnostics, drug delivery, and cell biology. In the present review, we first introduce standard methods for droplet generation (i.e., passive and active methods) and discuss the latest examples of emulsification and particle synthesis approaches enabled by microfluidic platforms. Then, the applications of droplet-based microfluidics in different biomedical applications are detailed. Finally, a general overview of the latest trends along with the perspectives and future potentials in the field are provided.

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