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iDiagnostics is the future of precision medicine. With your help we will make iDiagnostics available for all. Additionally, iDiagnostics will help to minimize the risk of humanity self-eradication. iDiagnostics employs TIRF microarray technology; to learn more about TIRF visit  www.tirf-labs.com

Handheld Molecular iDiagnostics

by TIRF Labs

www.tirf-labs.com

Mail: info@tirf-labs.com

Literature (Selected publications of our customers and our own data generated by TIRF Labs' products)

1.  Asanov A. “TIRF-EC Biosensors Massively parallel DNA and Protein Microarrays for Accurate and Rapid Detection of Pathogens.”  Biodetection Technologies 2007. Knowledge Press, Brookline, MA. 4th Edition,  2007, Chapter 17, p. 373-396, isbn # 1-59430-126-3.

2.  Asanov A. “TIRF-EC Biosensors with Reagentless Bioassays for Rapid and Accurate Detection of Pathogens.”  Biodetection Technologies 2008. Knowledge Press, Brookline, MA. 5th Edition, 2008, Chapter 8, p. 155-190, isbn # 1-59430-137-9.

3. Asanov A., Wilson W., and Oldham P. “Regenerable biosensor using total internal reflection fluorescence with electrochemical control”. US Patent 6,511,854. Publication Date: 2003-01-28.  (Active patent.)

4. Asanov A., Zepeda A., Vaca L. “Combined DNA and Protein Microarrays Based on Total Internal Reflection Fluorescence.”  Sensors, 2012, 12, 1800.

5. Patents pending 2016-2017.  

6. Willoughby D., Everett K., Halls M., Pacheco J., Skroblin P., Vaca L., Klussmann E., and Cooper D.  “Direct Binding Between Orai1 and AC8 Mediates Dynamic Interplay Between Ca2+ and cAMP Signaling.” Science Signaling, 2012, 5 (219), 29.

7. Sampieri A., Zepeda A., Asanov A., and Vaca L. “Visualizing the store operated channel complex assembly in real time: Identification of SERCA2 as a new member.” Cell Calcium, 2009, 45, 439.

8. Asanov A., Zepeda A., and Vaca L. “A novel form of Total Internal Reflection Fluorescence Microscopy (LG-TIRFM) reveals different and independent lipid raft domains in living cells.“ Biochim. Biophys. Acta, 2010, 1801, 147.

9. Asanov A, Sherry R, Sampieri A, and Vaca L. “A relay mechanism between EB1 and APC facilitate STIM1 puncta assembly at endoplasmic reticulum plasma membrane junctions”. Cell Calcium, 2013, 54, 246– 256

10. Luz-Madrigal A, Asanov A, Camacho-Zarco AR, Sampieri A, and Vaca L.  “A Cholesterol Recognition Amino Acid Consensus Domain in GP64 Fusion Protein Facilitates Anchoring of Baculovirus to Mammalian Cells”.  Journal of Virology, 2013, 87 (2), 11894–11907.

11. Additional publications of our customers with data generated by our TIRF and TIRF-EC products are available at the URL: www.tirf-labs.com

iDiagnostics Application Development Kit

and uTIRF Biodetection Station

iDiagnostics is a novel molecular diagnostics platform, which employs the technology of real-time TIRF microarray. The platform is supersensitive, accurate,  rapid, and yet affordable for home use. If you are developing fluorescence, electro-chemi-luminescence (ECL), or bioluminescence bioassays for diagnosing or prognosing, we invite you to collaborate. We will help you to interface your assays with the iDiagnostics platform and will be delighted to license your technology. Ask about uTIRF and the iDiagnostics Application Development Kit (ADK).

iDiagnostics simultaneously detects protein, nucleic acid, and metabolite biomarkers. It requires no or minimal sample preparation and is capable of detecting from one to several thousands of molecular markers in a 50-microliter sample of biological fluids, including whole blood. The high sensitivity and broad dynamic range of iDiagnostics covers the entire spectrum of clinically significant concentrations. The Limit of Detection (LOD) for micro-RNA is   ~10-18M.  For proteins and metabolites the LOD depends on the assay; for certain antibody-based assays LOD is  ~10-15M.  

Classical TIRF microarrays operate with small, sub-monolayer amounts of antibodies and DNA probes that are  immobilized on the surface; fluorescence signal is small, a low light photodetector, e.g. EMCCD camera is necessary. In iDiagnostics, 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 the usage of 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. iDiagnostics is ideal platform for interfacing antibody-based bioassays for detecting proteins and molecular beacon assays for measuring nucleic acids.  

The early detection of cancer, cardiovascular and other diseases, preventing pandemics, and other healthcare tasks require a highly sensitive, accurate, rapid, and yet affordable molecular diagnostics. Eleven years of research at TIRF Labs have resulted in the development of such a technology: a handheld platform, named iDiagnostics. It is a thousand times more accurate than traditional methods, is rapid and robust, and yet is inexpensive.

The goal of TIRF Labs is to provide iDiagnostics to every family on the globe. This goal cannot be achieved by a single company; we envision a collective global effort from the diagnostics community using the Open Innovation Business Model, which will stimulate a flux of intellectual property (IP) in and out of the iDiagnostics project. TIRF Labs will be delighted to license the IP of other groups and will offer the opportunity to license our own IP, in certain cases free of charge.

To facilitate all of the stages of this challenging project, from biomarker discovery, to clinical testing, TIRF Labs offers the uTIRF biodetection station and the Application Development Kit (ADK). Figs. 1 and 2 show the optical scheme and photos of uTIRF, the lightguide-based TIRF accessory, and manual microarrayer. Fig. 3 shows the iDiagnostics handheld cradle, and reusable cartridges designed for the development stage. Fig. 4 illustrates the stages of data acquisition and analysis, and Fig.5 - printing of TIRF microarrays by solid pin spotting. ADK-uTIRF products are available to all members of the diagnostic community, including academic, industrial, and government research groups. The NIH, FDA and CDC will be involved in the coordination of the collective efforts.

Two main products create the foundation for the collective project: (i) uTIRF - a multifunctional  biodetection station, which is sensitive down to single molecules and includes  lightguide-, prism-, and objective-based TIRF, a low light CMOS camera, three-color illuminator, and 1X, 4X, and 60X objectives, as well as (ii) the iDiagnostics ADK, which consists of the TIRF handheld cradle, cartridge blanks, manual microarrayer, and reagents kit. TIRF Labs has already received numerous requests from research groups worldwide to supply these products. In the beginning of 2019 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 iDiagnostics slides include assays for detecting protein, nucleic acid, metabolite  biomarkers. The arrays contain internal controls to ensure reliability and calibration. The cartridge is equipped with a 20-microliters flow cell, which encompasses the  microarray. iDiagnostics requires no or minimum sample preparation. Whole blood can be analyzed after the addition of an anticoagulant. 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

Fig. 5. Printing TIRF microarrays by solid pin spotting

Open Innovation Approach for collaborative development of molecular diagnostic applications

If you are developing a molecular diagnostics system based on fluorescence or electro-chemi-luminescence assays, we are offering to you an exciting opportunity to collaborate. We will help you with unique hardware, software, cartridge blanks, development tools, reagent kits, methods and protocols to facilitate the interfacing of your assays with advanced iDiagnostics platform. We will be delighted to license your Intellectual Property (IP) and offer you the opportunity to license our IP to develop precise and rapid, and yet affordable devices for molecular diagnosis and prognosis.  

The iDiagnostics platform provides unprecedented precision and rapid diagnosing due to its superior sensitivity, multiplicity, and fast response rate. The platform is several orders of magnitude more sensitive and more accurate than traditional methods. One of the advantages of iDiagnostics involves the simultaneous detection of multiple molecular markers of different classes, including proteins, nucleic acids, and metabolites. Detection of different classes of  markers is necessary for minimization of  the rate  of false positive responses.  

iDiagnostics employs 3D-encapsulated real-time TIRF microarrays that include internal controls and standards for normalization and calibration, which provides high reliability and broad dynamic range to iDiagnostics analyses. The platform also supports other luminescence assays, including Electro-Chemi-Luminescence (ECL). iDiagnostics integrates optics, microfluidics, electronics, electrochemical system, and nanoengineered assays into a small handheld device.  Real-time TIRF microarrays are capable of parallel detecting up to thousands of analytes, including proteins, nucleic acids, toxins and chemical agents.

iDiagnostics requires no or minimal sample preparation and can analyze complex biological fluids, including whole blood, saliva, and urine. It employs reagentless bioassays based on molecular beacons and sandwich-format antibody-based assays. Typically, molecular markers are detected in several  seconds or a few minutes after the sample is applied.

 Applications for the iDiagnostics platform include early diagnosing of cancer, heart diseases, STD, influenza, other infectious diseases, food and water safety, biodefense, forensic, military, environmental, and agricultural analyses.  

   Our goal is to make accurate iDiagnostics available to every one. Open Innovation Approach, which we use in this project, is well suited for collaborative development of advanced diagnostic products. If you are interested to become a part of this exciting project please send us an email at info@tirf-labs.com.

The scale of this project is very large. It is impossible for a single firm to address all of the numerous challenges. Several research groups have already joined the iDiagnostics project. Numerous other groups have expressed their interest to join. To progress rapidly this project requires significant funding. Therefore, we address our requests for funding to The Giving Pledge members and similarly positioned individuals, who are willing to give back to society. We believe that this project is giving a sensible and efficient opportunity of returning back to the society. Your help through donations, sponsorships, and grants will help  protect humanity, which today remains so fragile under the weight of many perils.

iDiagnostics will change the paradigm of medicine; it will relocate diagnosing from medical centers to individuals, make diagnosing accurate, rapid, minimally invasive, and affordable for all. It will help to prevent spreading of infectious diseases, enable personalized medicine, and prevent abuse of biotechnology.