IP Library › Granted Patent US 12,350,669
Granted Patent B2
US 12,350,669 · App. 18/542,574 · Granted Jul 8, 2025

Dual-depth thermoplastic microfluidic device and related systems and methods

Inventors: Rolf Muller (Del Mar, CA); Mateusz Hupert (Lawrence, KS)
Assignee: BioFluidica, Inc.
B01L3/502753B01L3/021B01L3/502715B01L2200/027B01L2200/0647B01L2200/12B01L2300/0816B01L2300/12B01L2400/0487B01L2400/086
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Quick Facts
Patent No.
US 12,350,669
App. No.
18/542,574
Granted
Jul 8, 2025
Kind
B2
Abstract

The presently disclosed subject matter provides dual-depth thermoplastic microfluidic devices, related kits, microfluidic systems comprising the dual-depth thermoplastic microfluidic device, methods of isolating nucleic acid analytes from a liquid sample, and methods of isolating extracellular vesicles from a liquid sample.

Claims (59)

1. A dual-depth thermoplastic microfluidic device comprising:

a thermoplastic substrate comprising an inlet microchannel, an outlet microchannel, and one or more isolation beds comprising a plurality of microposts,

wherein the one or more isolation beds are connected to the inlet microchannel and outlet microchannel;

wherein a cross-section of each of the inlet microchannel, the outlet microchannel, and the microposts each has a height and width,

wherein the inlet microchannel height and the outlet microchannel height are each greater than the height of the microposts, and

wherein the inlet microchannel, the outlet microchannel, and the one or more isolation beds are a single dual-depth fluidic layer.

2. The dual-depth thermoplastic microfluidic device of claim 1 , wherein the thermoplastic substrate is cyclic olefin copolymer (COC), cyclic olefin polymer (COP), polycarbonate (PC), polymethylmethacrylate, (PMMA), polystyrene (PS), polyvinylchloride (PVC), or polyethyleneterephthalate glycol (PETG).

3. The dual-depth thermoplastic microfluidic device of claim 1 , wherein the microposts comprise capture elements.

4. The dual-depth thermoplastic microfluidic device of claim 3 , wherein the capture elements are antibodies, antigen binding fragments of antibodies, or aptamers.

5. The dual-depth thermoplastic microfluidic device of claim 3 , wherein the capture elements are surface-bound oxygen-rich moieties such as carboxylic acid groups, salicylates, or esters.

6. The dual-depth thermoplastic microfluidic device of claim 1 , wherein the microposts are UV-activated.

7. The dual-depth thermoplastic microfluidic device of claim 1 , wherein the microposts are UV/O 3 -activated.

8. A kit comprising the dual-depth thermoplastic microfluidic device of claim 1 , and at least one reagent or buffer for use in processing a liquid sample using the dual-depth thermoplastic microfluidic device.

9. A microfluidic system comprising:

the dual-depth thermoplastic microfluidic device of claim 1 , wherein the dual-depth thermoplastic microfluidic device further comprises an inlet port in fluid communication with an outlet port;

a first automated pipetting channel comprising a first pump, and a first pipette tip coupled to the inlet port;

a second automated pipetting channel comprising a second pump, and a second pipette tip coupled to the outlet port; and

a non-transitory computer readable medium in communication with the first pump and the second pump, and programmed to command the first pump of the first automated pipetting channel and the second pump of the second automated pipetting channel to control flow of a liquid through the dual-depth thermoplastic microfluidic device.

10. A method of isolating nucleic acid analytes from a liquid sample comprising:

providing the dual-depth thermoplastic microfluidic device of claim 1 , wherein the microposts comprise capture elements that selectively bind a nucleic acid analyte;

controlling flow of a liquid sample through the dual-depth thermoplastic microfluidic device; and

binding the nucleic acid analyte to the capture elements thereby isolating the nucleic acid analytes from the liquid sample.

11. The method of claim 10 , wherein the dual-depth thermoplastic microfluidic device further comprises an inlet port in fluid communication with an outlet port; and

The method further comprises providing a system to control flow of the liquid sample through the dual-depth thermoplastic microfluidic device, wherein the system comprises:

a first automated pipetting channel comprising a first pump, and a first pipette tip coupled to the inlet port;

a second automated pipetting channel comprising a second pump, and a second pipette tip coupled to the outlet port; and

a non-transitory computer readable medium in communication with the first pump and the second pump, and programmed to command the first pump of the first automated pipetting channel and the second pump of the second automated pipetting channel to control flow of a liquid through the dual-depth thermoplastic microfluidic device.

12. The method of claim 10 , wherein the nucleic acid analytes are cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), genomic DNA (gDNA), or RNA.

13. The method of claim 10 , wherein the capture elements are surface-bound carboxylic acid groups, and the method comprises controlling flow of the liquid sample mixed with an immobilization buffer through the dual-depth thermoplastic microfluidic device.

14. The method of claim 10 , wherein the liquid sample is blood or any fraction or component thereof, cerebrospinal fluids, urine, sputum, saliva, pleural effusion, stool and seminal fluid.

15. The method of claim 10 , wherein the liquid sample is plasma.

16. The method of claim 10 , wherein >80% or >90% of nucleic acid fragments 50-750 bp in size are isolated and recovered.

17. The method of claim 10 , wherein >70% of nucleic acid fragments 50-750 bp in size are isolated and recovered.

18. A method of isolating extracellular vesicles from a liquid sample comprising:

providing the dual-depth thermoplastic microfluidic device of claim 1 , wherein the microposts comprise capture elements that selectively bind extracellular vesicles;

controlling flow of a liquid sample through the dual-depth thermoplastic microfluidic device; and

binding the extracellular vesicles to the capture elements thereby isolating the extracellular vesicles from the liquid sample.

19. The method of claim 18 , wherein the dual-depth thermoplastic microfluidic device further comprises and inlet port in fluid communication with an outlet port; and

the method further comprises providing a system to control flow of the liquid sample through the dual-depth thermoplastic microfluidic device, wherein the system comprises:

a first automated pipetting channel comprising a first pump, and a first pipette tip coupled to the inlet port;

a second automated pipetting channel comprising a second pump, and a second pipette top coupled to the outlet port; and

a non-transitory computer readable medium in communication with the first pump and the second pump, and programmed to command the first pump of the first automated pipetting channel and the second pump of the second automated pipetting channel to control flow of a liquid through the dual-depth thermoplastic microfluidic device.

20. The method of claim 18 , wherein the extracellular vesicles are exosomes.

21. The method of claim 18 , wherein the capture elements are antibodies, antigen binding fragments of antibodies, or aptamers.

22. The method of claim 18 , wherein the capture elements are monoclonal antibodies.

23. The method of claim 18 , wherein the capture elements are immobilized to the microposts by a single-stranded oligonucleotide bifunctional cleavable linker, or a photocleavable linker.

24. The method of claim 18 , wherein the capture elements are immobilized to the microposts via surface-bound carboxylic acid groups.

25. The method of claim 18 , wherein the liquid sample is blood or any fraction or component thereof, bone marrow, pleural fluid, peritoneal fluid, cerebrospinal fluid, urine, saliva, amniotic fluid, ascites, broncho-alveolar lavage fluid, synovial fluid, breast milk, sweat, tears, joint fluid, and bronchial washes.

26. The method of claim 18 , wherein the liquid sample is plasma.

27. The method of claim 18 , wherein the method further comprises lysis of the extracellular vesicles, RNA purification, RNA extraction, reverse transcription, and mRNA expression profiling.

28. The method of claim 18 , wherein the method further comprises obtaining distinct mRNA profiles indicative of the phenotype of the cells from which the extracellular vesicles originated.

29. The method of claim 18 , wherein the method further comprises release of the extracellular vesicles and nanoparticle tracking analysis.

30. The method of claim 18 , wherein the method further comprises release of the extracellular vesicles and transmission electron microscopy analysis.

31. The dual-depth thermoplastic microfluidic device of claim 1 further comprising one or more microchannels, and

wherein the one or more isolation beds are connected to the inlet microchannel and outlet microchannel by the one or more microchannels, and

wherein the inlet microchannel, the outlet microchannel, the one or more microchannels, and the one or more isolation beds are a single dual-depth fluidic layer.

32. The dual-depth thermoplastic microfluidic device of claim 31 ,

wherein a cross-section of each of the one or more microchannels each has a height and width, and

wherein the inlet microchannel height and the outlet microchannel height are each greater than the height of the one or more microchannels.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 10, 2024
From: MULLER, ROLF; HUPERT, MATEUSZ
To: BIOFLUIDICA, INC.
Reel/Frame 067949/0562 →
Continuity (4)
Continuation 18079860 · Dec 12, 2022
Continuation PCTUS2021037297 · Jun 14, 2021
Provisional Application 63038492 · Jun 12, 2020
Related Publication 20240351034A1 · Oct 24, 2024
References Cited (147)
US 6193471B1 · Paul · 2001 [cited by applicant]
US 6685668B1 · Cho et al. · 2004 [cited by applicant]
US 6869571B2 · Ingenhoven et al. · 2005 [cited by applicant]
US 6915679B2 · Chien et al. · 2005 [cited by applicant]
US 7858040B2 · Okun et al. · 2010 [cited by applicant]
US 8287820B2 · Williams et al. · 2012 [cited by applicant]
US 8394645B2 · Beebe et al. · 2013 [cited by applicant]
US 8425840B2 · Hosokawa et al. · 2013 [cited by applicant]
US 8470246B2 · Rich · 2013 [cited by applicant]
US 8557570B2 · Sahoo et al. · 2013 [cited by applicant]
US 9081001B2 · Cook et al. · 2015 [cited by applicant]
US 9339815B2 · Kim et al. · 2016 [cited by applicant]
US 9375531B1 · Lee et al. · 2016 [cited by applicant]
US 9500664B2 · Ness et al. · 2016 [cited by applicant]
US 9592501B2 · Jarvius et al. · 2017 [cited by applicant]
US 10393726B2 · Soper et al. · 2019 [cited by applicant]
US 11865541B2 · Muller · 2024 [cited by examiner]
US 20020100714A1 · Staats · 2002 [cited by applicant]
US 20030236489A1 · Jacobson et al. · 2003 [cited by applicant]
US 20040265171A1 · Pugia · 2004 [cited by examiner]
US 20060193730A1 · Rosenstein et al. · 2006 [cited by applicant]
US 20060257290A1 · Shimizu · 2006 [cited by applicant]
US 20070054293A1 · Liu et al. · 2007 [cited by applicant]
US 20080131323A1 · Kuczenski et al. · 2008 [cited by applicant]
US 20090014360A1 · Toner et al. · 2009 [cited by applicant]
US 20090074637A1 · Murphy et al. · 2009 [cited by applicant]
US 20090229979A1 · Champagne · 2009 [cited by applicant]
US 20100291584A1 · Tseng et al. · 2010 [cited by applicant]
US 20120100521A1 · Soper et al. · 2012 [cited by applicant]
US 20120128538A1 · Miller et al. · 2012 [cited by applicant]
US 20120244043A1 · Leblanc et al. · 2012 [cited by applicant]
US 20130078733A1 · Holmes et al. · 2013 [cited by applicant]
US 20140030811A1 · Cao et al. · 2014 [cited by applicant]
US 20140238122A1 · Mostowfi et al. · 2014 [cited by applicant]
US 20150251181A1 · Saito · 2015 [cited by applicant]
US 20160289669A1 · Fan et al. · 2016 [cited by applicant]
US 20160339431A1 · Shmilovich et al. · 2016 [cited by applicant]
US 20170043340A1 · West et al. · 2017 [cited by applicant]
US 20180272356A1 · Valerio · 2018 [cited by applicant]
US 20190219578A1 · Mitsuhashi · 2019 [cited by examiner]
US 20200023351A1 · Muller et al. · 2020 [cited by applicant]
CN 105899954A · 2016 [cited by applicant]
CN 105950469A · 2016 [cited by applicant]
EP 1206966A1 · 2002 [cited by applicant]
EP 1712285A1 · 2006 [cited by applicant]
JP 2011512125A · 2011 [cited by applicant]
JP 2013524171A · 2013 [cited by applicant]
JP 2015166707A · 2015 [cited by applicant]
WO 2009069656A1 · 2009 [cited by applicant]
WO 2012056369A1 · 2012 [cited by applicant]
WO 2016024941A1 · 2016 [cited by applicant]
WO 2016118915A1 · 2016 [cited by applicant]
WO 2016201163A1 · 2016 [cited by applicant]
WO WO2018200652A1 · 2018 [cited by applicant]
International Search Report for Application No. PCT/US2021/037297 dated Oct. 7, 2021, 6 pages. [cited by applicant]
Written Opinion for Application No. PCT/US2021/037297 dated Oct. 7, 2021, 6 pages. [cited by applicant]
Adamski, Mateusz, et al., “CD15+ Granulocyte and CD8+ T Lymphocyte Based Gene Expression Clusters for Ischemic Stroke Detection”, Medical Research Archives, vol. 5, Issue 11, Nov. 2017, 13 pages. [cited by applicant]
Ashcroft, B.A., et al., “Determination of the Size Distribution of Blood Microparticles Directly in Plasma Using Atomic Force Microscopy and Microfluidics”, Biomed Microdevices, Issue 14, Mar. 6, 2012, pp. 641-649. [cited by applicant]
Bell, George I., “Models for the Specific Adhesion of Cells to Cells”, Science, vol. 200, Issue 4342, May 12, 1978, pp. 618-627. [cited by applicant]
Boenisch, Thomas, “Handbook, Immunochemical Staining Methods”, DAKO Corporation, 2001, 67 pages. [cited by applicant]
Chalela, Julio A., M.D., et al., “Magnetic Resonance Imaging and Computed Tomography in Emergency Assessment of Patients with Suspected Acute Stroke: A Prospective Comparison”, The Lancet, vol. 369, Jan. 27, 2007, pp. 2… [cited by applicant]
Chang, Kai-Chien, et al., “The Forward Rate of Binding of Surface-Tethered Reactants: Effect of Relative Motion Between Two Surfaces”, Biophysical Journal, vol. 76, Mar. 1999, pp. 1280-1292. [cited by applicant]
Chen, Chihchen, et al., “Microfluidic Isolation and Transcriptome Analysis of Serum Microvesicles”, Lab Chip, vol. 10, Issue 4, Feb. 21, 2010, pp. 505-511. [cited by applicant]
Chen, Claire C., et al., “Elucidation of Exosome Migration Across the Blood-Brain Barrier Model in Vitro”, Cellular and Molecular Bioengineering, vol. 9, Issue 4, Dec. 2016, pp. 50-529. [cited by applicant]
Cho, Siwoo, et al., “Isolation of Extracellular Vesicle from Blood Plasma Using Electrophoretic Migration Through Porous Membrane”, Sensors and Actuators: B Chemical, Issue 233, Oct. 5, 2016, pp. 289-297. [cited by applicant]
Contreras-Naranjo, Jose C., et al., “Microfluidics for Exosome Isolation and Analysis: Enabling Liquid Biopsy for Personalized Medicine”, Lab Chip, Issue 17, Aug. 10, 2017, pp. 3558-3577. [cited by applicant]
Davies, Ryan T., et al., “Microfluidic Filtration System to Isolate Extracellular Vesicles from Blood”, Lab on a Chip, Issue 12, Nov. 13, 2012, pp. 5202-5210. [cited by applicant]
Deun, Van J., et al., “EV-Track: Transparent Reporting and Centralizing Knowledge in Extracellular Vesicle Research”, Nature Methods, vol. 14, No. 3, Mar. 2017, pp. 228-232. [cited by applicant]
Dreyer, Rachel, PhD, et al., “Most Important Outcomes Research Papers on Stroke and Transient Ischemic Attack”, Circulation: Cardiovascular Quality and Outcomes, vol. 7, Issue 1, Jan. 2014, pp. 191-204. [cited by applicant]
Eitan, Erez, et al., Extracellular Vesicle-Depleted Fetal Bovine and Human Sera Have Reduced Capacity to Support Cell Growth, Journal of Extracellular Vesicles, Issue 4, Mar. 26, 2015, 10 pages. [cited by applicant]
Fang, Shimeng, et al., “Clinical Application of a Microfluidic Chip for Immunocapture and Quantification of Circulating Exosomes to Assist Breast Cancer Diagnosis and Molecular Classification”, PLOS ONE, vol. 12, Issue … [cited by applicant]
Fonarow, Gregg C., et al., “Door-to-Needle Times for Tissue Plasminogen Activator Administration and Clinical Outcomes in Acute Ischemic Stroke Before and After a Quality Improvement Initiative”, JAMA, vol. 311, Issue 1… [cited by applicant]
Geis-Asteggiante, Lucia, et al., “Differential Content of Proteins, mRNAs, and miRNAs Suggests that MDSC and Their Exosomes May Mediate Distinct Immune Suppressive Functions”, Journal of Proteome Research, vol. 17, Issu… [cited by applicant]
Greenberg, James M., et al., “Immunophenotypic and Cytogenetic Analysis of Molt-3 and Molt-4: Human T-Lymphoid Cell Lines with Rearrangement of Chromosome 7”, Blood, vol. 72, No. 5, Nov. 1988, pp. 1755-1760. [cited by applicant]
He, Mei, et al., “Integrated Immunoisolation and Protein Analysis of Circulating Exosomes Using Microfluidic Technology”, Lab Chip, vol. 14, Issue 19, Oct. 7, 2014, pp. 3773-3780. [cited by applicant]
Hsu, Hsien-Yeh, et al., “Lipopolysaccharide-Mediated Reactive Oxygen Species and Signal Transduction in the Regulation of Interleukin-1 Gene Expression”, The Journal of Biological Chemistry, vol. 277, No. 25, Jun. 21, 2… [cited by applicant]
Huang, Tao, et al, “Current Progresses of Exosomes as Cancer Diagnostic and Prognostic Biomarkers”, International Journal of Biological Sciences, vol. 15, Issue 1, Jan. 6, 2019, 11 pages. [cited by applicant]
Im, Hyungsoon, et al., “Label-Free Detection and Molecular Profiling of Exosomes with a Nano-Plasmonic Sensor”, Nature Biotechnology, vol. 32, Issue 5, May 2014, pp. 490-495. [cited by applicant]
Jackson, Joshua M., et al., “Materials and Microfluidics: Enabling the Efficient Isolation and Analysis of Circulating Tumour Cells”, Chemical Society Reviews, vol. 46, Issue 14, Jul. 17, 2017, pp. 4245-4280. [cited by applicant]
Jackson, Joshua M., et al., “UV Activation of Polymeric High Aspect Ratio Microstructures: Ramifications in Antibody Surface Loading for Circulating Tumor Cell Selection”, Lab Chip, vol. 14, Issue 1, Jan. 7, 2014, pp. 1… [cited by applicant]
Jauch, Edward C., et al., “Guidelines for the Early Management of Patients with Acute Ischemic Stroke”, Stroke, vol. 44, Issue 3, Mar. 2013, pp. 870-947. [cited by applicant]
Ji, Qiuhong, et al., “Increased Brain-Specific MiR-9 and MiR-124 in the Serum Exosomes of Acute Ischemic Stroke Patients”, PLoS ONE, vol. 11, Issue 9, Sep. 23, 2016, 14 pages. [cited by applicant]
Kalafut, Mary A., et al., “Detection of Early CT Signs of >1/3 Middle Cerebral Artery Infarctions”, Stroke, vol. 31, Issue 7, Jul. 2000, pp. 1667-1671. [cited by applicant]
Kanwar, Shailender Singh, et al., “Microfluidic Device (ExoChip) for On-Chip Isolation, Quantification and Characterization of Circulating Exosomes”, Lab Chip, vol. 14, Issue 11, Jun. 7, 2014, pp. 1891-1900. [cited by applicant]
Ko, Jina, et al., “Smartphone-Enabled Optofluidic Exosome Diagnostic for Concussion Recovery”, Scientific Reports, Issue 6, Article No. 31215, Aug. 8, 2016, 12 pages. [cited by applicant]
Labat-Gest, Vivien, et al., “Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical esion Model for Mouse Stroke Studies”, Journal of Visualized Experiments, Issue 76, Jun. 9, 2013, 6 pag… [cited by applicant]
Lee, Kyungheon, et al., “Acoustic Purification of Extracellular Microvesicles”, ACS Nano, vol. 9, Issue 3, Mar. 24, 2015, pp. 2321-2327. [cited by applicant]
Liang, Li-Guo, et al., “An Integrated Double-Filtration Microfluidic Device for Isolation, Enrichment and Quantification of Urinary Extracellular Vesicles for Detection of Bladder Cancer”, Scientific Reports, Issue 7, A… [cited by applicant]
Liu, Chao, “Field-Free Isolation of Exosomes from Extracellular Vesicles by Microfluidic Viscoelastic Flows”, ACES Nano, vol. 11, Issue 7, Jul. 5, 2017, pp. 6968-6976. [cited by applicant]
Nair, Soumya V., et al., “Enzymatic Cleavage of Uracil-Containing Single-Stranded DNA Linkers for the Efficient Release of Affinity-Selected Circulating Tumor Cells”, Chemical Communications, Jan. 7, 2015, pp. 3266-3269. [cited by applicant]
Pahattuge, Thilanga N., et al., “Visible Photorelease of Liquid Biopsy Markers Following Microfluidic Affinity-Enrichment”, Chemical Communications, vol. 56, Issue 29, Apr. 14, 2020, pp. 4098-4101. [cited by applicant]
Prat, Aleix, et al., “Characterization of Cell Lines Derived from Breast Cancers and Normal Mammary Tissues for the Study of the Intrinsic Molecular Subtypes”, Breast Cancer Research and Treatment, Issue 142, Oct. 27, 2… [cited by applicant]
Reátegui, Eduardo, et al., “Engineered Nanointerfaces for Microfluidic Isolation and Molecular Profiling of Tumor-Specific Extracellular Vesicles”, Nature Communications, vol. 9, Issue 1, Jan. 12, 2018, 11 pages. [cited by applicant]
Rezeli, Melinda, et al., “Comparative Proteomic Analysis of Extracellular Vesicles Isolated by Acoustic Trapping or Differential Centrifugation”, vol. 88, No. 17, Aug. 3, 2016, pp. 8577-8586. [cited by applicant]
Rider, Mark A., et al., “ExtraPEG: A Polyethylene Glycol-Based Method for Enrichment of Extracellular Vesicles”, Scientific Reports, Issue 6, Article 23978, Apr. 12, 2016, 14 pages. [cited by applicant]
Rothermundt, Matthias, et al., “S100B in Brain Damage and Neurodegeneration”, Microscopy Research and Technique, vol. 60, Issue 6, Apr. 2003, pp. 614-632. [cited by applicant]
Santana, Steven M., et al., “Microfluidic Isolation of Cancer-Cell-Derived Microvesicles from Hetergeneous Extracellular Shed Vesicle Populations”, Biomedical Microdevices, vol. 16, Issue 6, Dec. 2014, pp. 869-877. [cited by applicant]
Selvaraj, Uma Maheswari, et al., “Long-Term T Cell Responses in the Brain After an Ischemic Stroke”, Discovery Medicine, vol. 24, Issue 134, Dec. 2017, pp. 323-333. [cited by applicant]
Shao, Huilin, et al., “Chip-Based Analysis of Exosomal mRNA Mediating Drug Resistance in Glioblastoma”, Nature Communications, May 11, 2015. [cited by applicant]
Shelke, Ganesh Vilas, et al, “Importance of Exosome Depletion Protocols to Eliminate Functional and RNA-Containing Extracellular Vesicles from Fetal Bovine Serum”, Journal of Extracellular Vesicles, Issue 3, Sep. 30, 20… [cited by applicant]
Soper, Steven A., et al., “Polymeric Micoelectromechanical”, Analytical Chemistry, vol. 72, Oct. 1, 2000, pp. 642A-651A. [cited by applicant]
Tang, Yang, et al., “Gene Expression in Blood Changes Rapidly in Neutrophils and Monocytes After Ischemic Stroke in Humans: a Microarray Study”, Journal of Cerebral Blood Flow & Metabolism, vol. 26, Issue 8, Aug. 2006, … [cited by applicant]
Théry, Clotilde, et al., “Isolation and Characterization of Exosomes from Cell Culture Supernatants and Biological Fluids”, Supplement 30, Unit 3.22, pp. 3.22.1-3.22.29. [cited by applicant]
Tough, David F., et al., “T Cell Stimulation in Vivo by Lipopolysaccharide (LPS)”, Journal of Experimental Medicine, vol. 185, No. 12, Jun. 16, 1977, pp. 2089-2094. [cited by applicant]
Van Deun, Jan, “EV-Track: Transparent Reporting and Centralizing Knowledge in Extracellular Visicle Research”, Nature Methods, vol. 14, No. 3, Mar. 2017, pp. 228-232. [cited by applicant]
Wang, Zongxing, et al., “Ciliated Micropillars for the Microfluidic-Based Isolation of Nanoscale Lipid Vesicles”, Lab on a Chip, Issue 15, Aug. 7, 2013, pp. 2879-2882. [cited by applicant]
BlackHole Lab, “What are the Different Thermoplastic Polymers Used in Microfluidics”, www.blackholelab-soft-lithography.com/different-thermoplastic-polymers-used-in-microfluidics, printed Nov. 26, 2019, 15 pages. [cited by applicant]
Brown, Virginia, “Development of an Extracellular Vesicle Microfluidic Affinity Purification (EV-MAP) in Vitro Assay for Breast Cancer,” Thesis submitted to the graduate degree program in Bioengineering of the Universit… [cited by applicant]
Campos, Camila D., et al, “Microfluidic-Based Solid Phase Extraction of Cell Free DNA” Electronic Supplemental Information, Lab Chip, Issue 22, Nov. 6, 2018, 11 pages. [cited by applicant]
Campos, Camila D., et al, “Microfluidic-Based Solid Phase Extraction of Cell Free DNA”, Lab Chip, Issue 22, Nov. 6, 2018, pp. 3459-3470. [cited by applicant]
Campos, Camila D.M., et al., “Molecular Profiling of Liquid Biopsy Samples for Precision Medicine”, The Cancer Journal, vol. 24, Issue 2, Mar./Apr. 2018, pp. 93-103. [cited by applicant]
Dynamic Devices, “Liquid Handling Robotics”, printed on Feb. 28, 2017, 8 pages. [cited by applicant]
Dynamic Devices, “VVP On-The-Fly Pipetting Correction,” printed on Feb. 28, 2017, 3 pages. [cited by applicant]
Dynamic Devices, “VVP Real Time Transfer Validation,” printed on Feb. 28, 2017, 3 pages. [cited by applicant]
Elveflow, “How to Choose the Right Microfluidic Flow Control System?”, printed on Feb. 19, 2017, 17 pages. [cited by applicant]
Fiorini, Gina S., et al., “Disposable Microfluidic Devices: Fabrication, Function, and Application”, BioTechniques, vol. 38, No. 3, Mar. 2005, pp. 329-446. [cited by applicant]
Gale, Bruce K., et al., “A Review of Current Methods in Microfluidic Device Fabrication and Future Commercialization Prospects”, Inventions, No. 3: 60, Aug. 28, 2018, 25 pages. [cited by applicant]
Gao, Kexin, et al., “Ultra-Low-Cost Fabrication of Polymer-Based Microfluidic Devices with Diode Laser Ablation”, Biomedical Microdevices 21, 83, Aug. 15, 2019. [cited by applicant]
Gencturk, Elif, et al., “Advances in Microfluidic Devices Made from Thermoplastics Used in Cell Biology and Analyses”, Biomicrofluidics, No. 11(5), Oct. 24, 2017. [cited by applicant]
Hamilton Robotics, “Microlab STAR Line—Artificial Intelligence with Hamilton Robotics”, 2013, 36 pages. [cited by applicant]
Hou, Han Wei, et al., “Microfluidic Devices for Blood Fractionation”, Micromachines, No. 2(3), Dec. 2011, pp. 319-343. [cited by applicant]
Kamande, J.W., et al., “Modular Microsystem for the Isolation, Enumeration, and Phenotyping of Circulating Tumor Cells in Patients with Pancreatic Cancer”, Analytical Chemistry, No. 85, Oct. 2013, pp. 9092-9100. [cited by applicant]
Kamande, J.W., et al., “Modular Microsystem for the Isolation, Enumeration, and Phenotyping of Circulating Tumor Cells in Patients with Pancreatic Cancer”, Electronic Supplementary Information, Analytical Chemistry, No.… [cited by applicant]
Loiseau, Etienne, et al., “Microfluidic Study of Enhanced Deposition of Sickle Cells at Acute Corners”, Biophysical Journal, vol. 108, Jun. 2015, pp. 2623-2632. [cited by applicant]
Mair, Dieudonne A., et al., “Injection Molded Microfluidic Chips Featuring Integrated Interconnects”, Lab Chip, Issue 10, No. 6, Jul. 31, 2006, pp. 1346-2354. [cited by applicant]
Matellan, Carlos, et al., “Cost-Effective Rapid Prototyping and Assembly of Poly(methyl methacrylate) Microfluidic Devices”, Scientific Reports, No. 8(1), May 3, 2018. [cited by applicant]
Nagrath, Sunitha, et al., “Isolation of Rare Circulating Tumour Cells in Cancer Patients by Microchip Technology”, Nature, Dec. 20, 2007, pp. 1235-1239. [cited by applicant]
Nagrath, Sunitha, et al., “Isolation of Rare Circulating Tumour Cells in Cancer Patients by Microchip Technology”, Supplementary Information, Nature, Dec. 20, 2007, pp. 1235-1239. [cited by applicant]
Ramirez, Marcel I., et al., “Technical Challenges of Working with Extracellular Vesicles”, Nanoscale, No. 10, Issue 3, Jan. 21, 2018, pp. 881-906. [cited by applicant]
Rana, Ankit, et al., “Advancements in Microfluidic Technologies for Isolation and Early Detection of Circulating Cancer-Related Biomarkers”, Analyst, Issue 13, May 23, 2018, pp. 2971-2991. [cited by applicant]
Sadeghi, Sahl, et al., “A Simple, Bubble-Free Cell Loading Technique for Culturing Mammalian Cells on Lab-on-a-Chip Devices”, Royal Society of Chemistry, Feb. 28, 2017, 5 pages. [cited by applicant]
Tsao, Chia-Wen, et al., “Bonding of Thermoplastic Polymer Microfluidics”, Microfluidics and Nanofluidics, No. 6, Nov. 13, 2008. [cited by applicant]
Turhan, Aslihan, et al., “Effect of Intraluminal Pillars on Particle Motion in Bifurcated Microchannels”, In Vitro Cell Dev Biol Anim., No. 44(10), Sep. 20, 2008, pp. 426-433. [cited by applicant]
Xu, Zheyun, et al., “Microfluidic Technologies for cfDNA Isolation and Analysis”, Micromachines, No. 10(10), Oct. 3, 2019. [cited by applicant]
Yanai, Takuma, “Hydrodynamic Microparticle Separation Mechanism Using Three-Dimensional Flow Profiles in Dual-Depth and Asymmetric Lattice-Shaped Microchannel Networks”, Micromachines, No. 10(6), Jun. 25, 2019. [cited by applicant]
Wei, Zhiyun, et al., “Coding and Noncoding Landscape of Extracellular RNA Released by Human Glioma Stem Cells”, Nature Communications, vol. 8, Article 1145, Oct. 26, 2017, 15 pages. [cited by applicant]
Willyard, Cassandra, “New Human Gene Tally Reignites Debate”, Nature, vol. 558, Jun. 21, 2018, pp. 354-355. [cited by applicant]
Witek, Malgorzata A., et al., “Purification and Preconcentration of Genomic DNA from Whole Cell Lysates Using Photoactivated Polycarbonate (PPC) Microfluidic Chips”, Nucleic Acids Research, vol. 34, Nov. 10, Jun. 6, 200… [cited by applicant]
Woo, Hyun-Kyung, et al., “Exodisc for Rapid, Size-Selective, and Efficient Isolation and Analysis of Nanoscale Extracellular Vesicles from Biological Samples”, vol. 11, Issue 2, Jan. 9, 2017, pp. 1360-1370. [cited by applicant]
Yang, Junfa, et al., “Extracellular Vesicles as Carriers of Non0Coding RNAs in Liver Diseases”, Frontiers in Pharmacology, vol. 9, Article 415, Apr. 24, 2018, 11 pages. [cited by applicant]
Yilmaz, Gokhan, et al., “Role of T Lymphocytes and Interferon-y in Ischemic Stroke”, Circulation, vol. 114, Issue 7, May 2, 2006, pp. 2105-2112. [cited by applicant]
Yoon, C.W., et al., “Premorbid Warfarin Use and Lower D-Dimer Levels are Associated with a Spontaneous Early Improvement in an Atrial Fibrillation-Related Stroke”, Journal of Thrombosis and Haemostasis, vol. 10, Issue 1… [cited by applicant]
Yoon, Yae Jin, et al., “Extracellular Vesicles as Emerging Intercellular Communicasomes”, BMB Reports, vol. 47, Issue 10, Jul. 18, 2014, pp. 531-539. [cited by applicant]
Yoshioka, Yusuke, et al., “Ultra-Sensitive Liquid Biopsy of Circulating Extracellular Vesicles Using ExoScreen”, Nature Communications, Issue 5, Article No. 3591, Apr. 7, 2014, 8 pages. [cited by applicant]
Zhang, Peng, et al., “Ultrasensitive Detection of Circulating Exosomes with a 3D-Nanopatterned Microfluidic Chip”, Nature Biomedical Engineering, vol. 3, Issue 6, Jun. 2019, pp. 438-451. [cited by applicant]
Zhang, Peng, et al., “Ultrasensitive Microfluidic Analysis of Circulating Exosomes Using a Nanostructured Graphene Oxide/Polydopamine Coating”, Lab on a Chip, Issue 16, Mar. 22, 2016, pp. 3033-3042. [cited by applicant]
Zhao, Zheng, et al., “A Microfluidic ExoSearch Chipo for Multiplexed Exosome Detection Towards Blood-Based Ovarian Cancer Diagnosis”, Lab on a Chip, Issue 16, 2016, pp. 489-496. [cited by applicant]