Development Tools: Handheld i-Diagnostics (Prototype) and Cartridge Blanks
In the framework of Open Innovation Business Model, TIRF Labs offers the prototype of the handheld i-Diagnostics device with cartridge blanks and sample preparation modules as development tools. This group of products is useful for the final steps of test development, including the interfacing of existing fluorescence bioassays with the i-Diagnostics platform.
If an existing set of assays and sample preparation procedures meet the criteria for fluorescence detection, assay immobilization and sample prep, early development stages can be omitted and the final development tasks can be performed with the handheld i-Diagnostics prototype and cartridge blanks (see pdf brochure). For example, the real-time TIRF microarray in i-Diagnostics can be used with the molecular beacon assay for the purpose of detecting of a COVID-19 RNA. In this case molecular beacons with a DNA sequence complementary to that of virus RNA are immobilized at the surface of the TIRF slide in the form of round spots (0.3 mm in diameter), encapsulated into silk fibroin hydrogel. Without the target RNA of the virus, fluorescence of the beacon is quenched. Upon binding the target RNA fluorescence dequenches, which means positive result for COVID-19. Similarly, a number of other applications, including Influenza, Ebola, SARS, STD, and other infectious diseases are ready for finalizing as the i-Diagnostics tests using cartridge blanks and standard sample prep modules. In such case, the i-Diagnostics test is a couple steps away from laboratory trials and several steps away from FDA approval. If new tests require procuring antibodies or other affinity reagents to build the bioassay, their subsequent calibration will be necessary for test development and validation, respectively.
If you are developing fluorescence, electro-chemi-luminescence (ECL), or bioluminescence bioassays for diagnosing or prognosing a disease or for other applications, we invite you to collaborate. Our team is here to assist you in interfacing your assays with i-Diagnostics platform and licensing your technology. Ask about uTIRF and i-Diagnostics Application Development Kit (ADK). Visit www.tirf-labs.com website to learn more about the Total Internal Reflection Fluorescence (TIRF) and the principles of real-time TIRF microarrays.
i-Diagnostics features a novel type of real-time microarray that simultaneously detects protein, DNA/RNA, and metabolite biomarkers. It requires no or minimal sample preparation and is capable of detecting a single molecule to several thousands of molecular markers in a 50-microliter sample of biological fluids, including whole blood. High sensitivity and the broad dynamic range of i-Diagnostics covers the entire spectrum of clinically significant concentrations. The Limit of Detection (LOD) for micro-RNA is ~10-18M. For proteins and metabolites LOD depends on the assay; for certain antibody-based assays LOD is ~10-15M.
Classical 2D TIRF microarrays operate with small, sub-monolayer amounts of antibodies and DNA probes immobilized on the surface; fluorescence signal is small, a low light photodetector, e.g. EMCCD camera is necessary. In i-Diagnostics the signal of TIRF arrays is enhanced by 3D encapsulation, which captures the excitation light and becomes an integral part of the lightguide. 3D encapsulation allows for using larger amounts of antibodies per unit area of bioassay spot. The signal from such arrays is a thousand-fold greater than that in classical arrays. CCD cameras of cellphones are sensitive enough to detect the signal. i-Diagnostics is ideal platform for interfacing antibody-based bioassays for detecting proteins and molecular beacon assays for measuring nucleic acids.
One of the unique features of this project is that i-Diagnostics is supplied with a family of tools and methods that facilitate all stages of the i-Diagnostics application development.
Fig. x shows the main components of the development tools: i-Diagnostics Application Development Kit (ADK) and ultimate-TIRF (uTIRF) Biodetection Station. Fig. x illustrates the main stages of microarray printing, assaying and data acquisition and analysis. Left panel of Fig. 1 shows an early prototype of i-Diagnostics and disposable cartridges.
Before the panels of assays are tested with i-Diagnostics cartridges and the cradle, there are several stages of assay development, validation, calibration and other tasks that require rapid and direct assess to microarray. These development procedures are implemented using uTIRF station. For the development purposes uTIRF is equipped with open perfusion chambers. There are several formats of open perfusion and closed chambers for different development tasks. Open perfusion chambers are suitable for easy access to each individual spot of the panel. For more information about i-Diagnostics ADK and uTIRF station refer to ADK and uTIRF pdf brochures and contact TIRF Labs via email: info@TIRF-Labs.com.
TIRF Labs is offering a family of products, tools, cartridge blanks, sample preparation modules, reagents, supplies, methods and protocols that facilitate all stages of innovations at i-Diagnostics platform. TIRF Labs’ role is to provide logistics and service support of these innovations and to shorten their way to FDA approvals and application by end-users. In practice, the combination of ADK products and services facilitates interfacing of existing assays with i-Diagnostics platform and stimulates the development of new i-Diagnostics applications. The main set of development products is shown in the Figure x. uTIRF station is shown with prism-TIRF accessory installed. The lightguide TIRF (lgTIRF) accessory is shown to the left from uTIRF. Cartridge blanks, manual microarrayer, and the handheld i-Diagnostics cradle with cartridge blanks from the Application Development Kit are shown at the left. These products have already been successfully used by several our collaborators.
i-Diagnostics Application Development Kit
i-Diagnostics Application Development Kit (ADK) is designed to facilitate all stages of assay development – from biomarker discovery to creating and testing the panels of assays. TIRF slide is the central part of i-Diagnostics cartridges. The slide carries on its surface microarrays that simultaneously detect protein, nucleic acid, and metabolite biomarkers. In most of the existing applications, protein, nucleic acid, and metabolite assays are enclosed into one common flow chamber. In specialized applications, protein, nucleic acid, and metabolite microarrays can be separated by partitions – enclosed into three parallel flow chambers. ADK is supplied with a manual microarrayer, which enables printing of the microarrays using micropipettes (Fig. x, microarrayer panel). Initial stages of bioassay development requires rapid access to individual spots of the bioassays. To facilitate the access, ADK contains open perfusion chambers (Fig. 1x lgTIRF panel) as opposed to closed flow cells that are necessary to measure fast kinetics of response (Fig. x, cartridges with a closed flow cell).
Fig. x shows the handheld i-Diagnostics cradle with cellphone inserted and cartridges for different applications. Fig. 5 Illustrates printing of i-Diagnostics microarrays by solid pin spotting using a robotic microarrayer. Fig. x illustrates the stages of data acquisition.
TIRF Labs has already received numerous requests from research groups worldwide to supply uTIRF and ADK products. We plan to start supplying these products along with experimental protocols. Several groups have already joined our project. We have incorporated their feedback and will incorporate future comments from our future customers in the next releases of the uTIRF and ADKs. We will supply advanced hardware, software, reagents, development tools, protocols, videos, and other support to accommodate the requirements for different diagnostic applications.
The arrays of bioassays printed at the surface of i-Diagnostics slides contain internal controls to ensure reliability and calibration. The cartridge is equipped with a 20-microliter flow cell, which encompasses the microarray. i-Diagnostics requires no or minimum sample preparation. Whole blood can be analyzed after the addition of an anticoagulant. A simple sample preparation module is included in the cartridge; space is reserved for more complex sample preparation modules as will be required for applications developed in the future. For more information contact us via email: info@TIRF Labs.com