Advection-enhanced kinetics in microtiter plates for improved surface assay quantitation and multiplexing capabilities

Abstracts Assessment of appropriate laboratory measurements to supplement the Crohn's disease activity index Hepatitis B surface antigen levels during the natural history of chronic hepatitis B: a perspective on Asia Hexokinase-2 bound to mitochondria: cancer's stygian link to the "Warburg Effect" and a pivotal target for effective therapy

Surface assays such as ELISA are pervasive in clinics and research and predominantly standardized in microtiter plates (MTP). MTPs provide many advantages but are often detrimental to surface assay efficiency due to inherent mass transport limitations. Microscale flows can overcome these and largely improve assay kinetics. However, the disruptive nature of microfluidics with existing labware and protocols has narrowed its transformative potential. We present WellProbe, a novel microfluidic concept compatible with MTPs. With it, we show that immunoassays become more sensitive at low concentrations (up to 9× signal improvement in 12x less time), richer in information with 3-4 different kinetic conditions, and can be used to estimate kinetic parameters, minimize washing steps and non-specific binding, and identify compromised results. We further multiplex single-well assays combining WellProbe’s kinetic regions with tailored microarrays. Finally, we demonstrate our system in a context of immunoglobulin subclass evaluation, increasingly regarded as clinically relevant.

Automation for Life Science Laboratories

The automation of processes in all areas of the life sciences will continue to increase in the coming years due to an ever increasing number of samples to be processed Gentaur Labware, an increasing need to protect laboratory personnel from infectious material and increasing cost pressure. Depending on the requirements of the respective application, different concepts for automation systems are available, which have a different degree of automation with regard to data handling, transportation tasks, and the processing of the samples.

Robots form a central component of these automation concepts. Classic stationary robots from the industrial sector will increasingly be replaced by new developments in the field of light-weight robots.
In addition, mobile robots will also be of particular importance in the automation of life science laboratories in the future, especially for transportation tasks between different manual and (partially) automated stations.
With an increasing number of different, highly diverse processes, the need for special devices and system components will also increase.
This applies to both, the handling of the labware and the processing of the samples. In contrast to previous automation strategies with a highly parallel approach, future developments will increasingly be characterized by individual sample handling.

Fluorescence-based Single-cell Analysis of Whole-mount-stained and Cleared Microtissues and Organoids for High Throughput Screening

Three-dimensional (3D) cell culture, especially in the form of organ-like microtissues (“organoids”), has emerged as a novel tool potentially mimicking human tissue biology more closely than standard two-dimensional culture. Typically, tissue sectioning is the standard method for immunohistochemical analysis. However, it removes cells from their native niche and can result in the loss of 3D context during analyses.

Automated workflows require parallel processing and analysis of hundreds to thousands of samples, and sectioning is mechanically complex, time-intensive, and thus less suited for automated workflows.
Here, we present a simple protocol for combined whole-mount immunostaining, tissue-clearing, and optical analysis of large-scale (approx. 1 mm) 3D tissues with single-cell level resolution.
While the protocol can be performed manually, it was specifically designed to be compatible with high-throughput applications and automated liquid handling systems.
This approach is freely scalable and allows parallel automated processing of large sample numbers in standard labware.
We have successfully applied the protocol to human mid- and forebrain organoids, but, in principle, the workflow is suitable for a variety of 3D tissue samples to facilitate the phenotypic discovery of cellular behaviors in 3D cell culture-based high-throughput screens.
Graphic abstract: Automatable organoid clearing and high-content analysis workflow and timeline.

Gold-Polyoxoborates Nanocomposite Prohibits Adsorption of Bacteriophages on Inner Surfaces of Polypropylene Labware and Protects Samples from Bacterial and Yeast Infections

Bacteriophages (phages) are a specific type of viruses that infect bacteria. Because of growing antibiotic resistance among bacterial strains, phage-based therapies are becoming more and more attractive. The critical problem is the storage of bacteriophages. Recently, it was found that bacteriophages might adsorb on the surfaces of plastic containers, effectively decreasing the titer of phage suspensions.

Here, we showed that a BOA nanocomposite (gold nanoparticles embedded in polyoxoborate matrix) deposited onto the inner walls of the containers stabilizes phage suspensions against uncontrolled adsorption and titer decrease. Additionally, BOA provides antibacterial and antifungal protection. The application of BOA assures safe and sterile means for the storage of bacteriophages.

Adsorption of bacteriophages on polypropylene labware affects the reproducibility of phage research

Hydrophobicity is one of the most critical factors governing the adsorption of molecules and objects, such as virions, on surfaces. Even moderate change of wetting angle of plastic surfaces causes a drastic decrease ranging from 2 to 5 logs of the viruses (e.g., T4 phage) in the suspension due to adsorption on polymer vials’ walls.

The effect varies immensely in seemingly identical containers but purchased from different vendors. Comparison of glass, polyethylene, polypropylene, and polystyrene containers revealed a threshold in the wetting angle of around 95°: virions adsorb on the surface of more hydrophobic containers, while in more hydrophilic vials, phage suspensions are stable.
The polypropylene surface of the Eppendorf-type and Falcon-type can accommodate from around 10⁸ PFU/ml to around 10¹⁰ PFU/ml from the suspension.
The adsorption onto the container’s wall might result in complete scavenging of virions from the bulk. We developed two methods to overcome this issue.
The addition of surfactant Tween20 and/or plasma treatment provides a remedy by modulating surface wettability and inhibiting virions’ adsorption.
Plastic containers are essential consumables in the daily use of many bio-laboratories.
Thus, this is important not only for phage-related research (e.g., the use of phage therapies as an alternative for antibiotics) but also for data comparison and reproducibility in the field of biochemistry and virology.

Resonant acoustic rheometry for non-contact characterization of viscoelastic biomaterials

Resonant Acoustic Rheometry (RAR) is a new, non-contact technique to characterize the mechanical properties of soft and viscoelastic biomaterials, such as hydrogels, that are used to mimic the extracellular matrix in tissue engineering. RAR uses a focused ultrasound pulse to generate a microscale perturbation at the sample surface and tracks the ensuing surface wave using pulse-echo ultrasound. The frequency spectrum of the resonant surface waves is analyzed to extract viscoelastic material properties.

In this study, RAR was used to characterize fibrin, gelatin, and agarose hydrogels. Single time point measurements of gelled samples with static mechanical properties showed that RAR provided consistent quantitative data and measured intrinsic material characteristics independent of ultrasound parameters. RAR was also used to longitudinally track dynamic changes in viscoelastic properties over the course of fibrin gelation, revealing distinct phase and material property transitions.

The application of RAR was verified using finite element modeling and the results were validated against rotational shear rheometry. Importantly, RAR circumvents some limitations of conventional rheology methods and can be performed in a high-throughput manner using conventional labware. Overall, these studies demonstrate that RAR can be a valuable tool to noninvasively quantify the viscoelastic mechanical properties of soft hydrogel biomaterials.

Micro Labware Kit
LA025-1NO	EWC Diagnostics	1 unit	16.29 EUR
Micro Labware Kit
LA025-5NO	EWC Diagnostics	1 unit	72.8 EUR
Gentodenz
19-DENZ-500	Gentaur Genprice	500 g	416 EUR
Rye Agar A
765-M1854-500G	Gentaur Genprice	500 g	82 EUR
Rye Agar B
765-M1855-500G	Gentaur Genprice	500 g	93 EUR
Porcine Parvovirus Antibody Elisa Test Kit
767-LSY-30009	Gentaur Genprice	192 Wells/kit	382 EUR
QuantiChrom Hemoglobin Assay Kit
65-DIHB-250	Gentaur Genprice	250 tests	473 EUR
Malachite Green Phosphate Assay kit
65-POMG-25H	Gentaur Genprice	2500 tests	333 EUR
50ml TC Tubes, Conical, 440 units/box
04-5540150	Gentaur Genprice	440 units/box	85.2 EUR
GMP IL4, 50µg
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SDS-Blue™ - Coomassie based solution for protein staining in SDS-PAGE
04-GSB	Gentaur Genprice	Ask for price Ask for price	1L 500mL
rHu IL 2 , 3MIU
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rHu IL 2 , 3MIU , Lot 200908F02
04-RHIL2-08F02	Gentaur Genprice	1 vial	298.8 EUR
PRE-GMP rHu GM-CSF, Molgramostim-Leukoma
04-RHUGM-CSF-7A10	Gentaur Genprice	300 µg	462 EUR
Mouse Anti TNP Immunotoxicity
198-TNPG-1	Gentaur Genprice	100 µL	469.2 EUR
Exo-Check Exosome Antibody Array
322-EXO-FBS-50A-1	Gentaur Genprice	100 µg	469.2 EUR
1-Step Polymorphs, Human Cell Separation
71-AN221725	Gentaur Genprice	1	238.8 EUR
Monkeypox Virus Real Time PCR Kit
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Monkeypox Virus Real Time PCR Kit
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GMP Recombinant Human Interleukin-4 (IL-4)
GMPhuIL-4-50ug	Gentaur Genprice	50ug	528 EUR
Purite Labwater 1 Deioniser Cartridge - EACH
WAT3314	Scientific Laboratory Supplies	EACH	176.85 EUR
Purite Labwater 2 Deioniser Cartridge - EACH
WAT3316	Scientific Laboratory Supplies	EACH	264.6 EUR
Purite Labwater 1 Water Deioniser with Coil and Gun - EACH
WAT3002	Scientific Laboratory Supplies	EACH	831.6 EUR
Purite Labwater 2 Water Deioniser with Coil and Gun - EACH
WAT3006	Scientific Laboratory Supplies	EACH	1026 EUR
Gentamycin (Gentamicin)
MBS600426-025mg	MyBiosource	0.25mg	580 EUR
Gentamycin (Gentamicin)
MBS600426-05mg	MyBiosource	0.5mg	965 EUR
Gentamycin (Gentamicin)
MBS600426-5x05mg	MyBiosource	5x0.5mg	4185 EUR
Gentamycin (Gentamicin) (AP)
MBS6124654-01mL	MyBiosource	0.1(mL	1100 EUR
Gentamycin (Gentamicin) (AP)
MBS6124654-5x01mL	MyBiosource	5x0.1mL	4805 EUR