Biochips to diagnose hemoglobin disorders and monitor blood cells
A technology of biochip and hemoglobin, applied in biological testing, disease diagnosis, biomaterial analysis, etc., can solve the problems of not being widely provided
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Embodiment 1
[0152] The HemeChip technology offers a novel and innovative solution to the challenges clinical facilities face in diagnosing SCD. In just ten minutes, the HemeChip uses cellulose acetate electrophoresis to separate less than 5 microliters of blood into bands showing the basic types of hemoglobin A, A2, S, C and F. Costing less than $2.00 per chip to run and manufacture, the Hemechip system is easy to use and transport. In addition, the chip's simple design is easy to mass produce. Hemechip technology provides an accurate, efficient, fast and inexpensive way to diagnose SCD.
[0153] Materials and methods
[0154] HemeChip material
[0155] 1.5 mm thick poly(methyl methacrylate) (PMMA) sheets were purchased from McMaster-Carr (Elmhurst, IL), and 1 / 32" thick PMMA sheets were purchased from ePlastics (San Diego, CA). 3M Optics Transparent double-sided adhesive (DSA) (type 8142) was purchased from iTapeStore (Scotch Plains, NJ). Prepared from 10× TBE buffer solution (Inv...
Embodiment 2
[0192] Microfluidic Biochip Fabrication
[0193] A PMMA top piece was prepared by cutting the inlet and outlet (0.61 mm diameter, 26 m apart) using a VersaLASER system (Universal Laser Systems Inc., Scottsdale, AZ). Double-sided adhesive (DSA) membranes (iTapestore, Scotch Plains, NJ) were cut to fit the dimensions of the PMMA sections and 28 x 4 mm channels. The DSA was then attached to the PMMA top section to include inlets and outlets between the contours of the DSA membrane. Gold-sealed glass slides were then assembled with PMMA-DSA structures to form microfluidic channels.
[0194] surface chemistry
[0195] Prepare a GMBS stock solution by dissolving 25 mg GMBS in 0.25 mL DMSO and dilute the stock solution with ethanol to obtain a 0.28% v / v GMBS working solution. FN was diluted with PBS to create a (1:10) FN working solution. Prepare a BSA solution by dissolving 3 mg of lyophilized BSA in 1 mL of PBS.
[0196] After assembly, the channels were washed with 30 μL ...
Embodiment 3
[0219] This example describes a microfluidic biochip comprising at least one etched microchannel comprising a fibronectin (FN) functionalized glass surface. The biochip also includes a poly(methyl methacrylate) plastic top with etched inlets and etched outlets. The top also includes a double-sided adhesive film in the middle that defines the profile and width (eg, approximately 50 μm) of the microchannel. ( Figure 9A and 13C). In addition, the microfluidic system can be placed on the OlympusIX83 inverted motorized microscope stage for high-resolution live single-cell image recording and analysis.
[0220] FN circulates in plasma and is present in endothelial cell membranes. FN is considered an adhesion glycoprotein that has been shown to play a role in the adhesion of HbSRBC to the endothelial wall. Kasschau et al., Blood (blood) 87(2): 771-780 (1996); Wick et al., Current opinion in hematology (current opinion in hematology) 3(2): 118-124 (1996); Mosher, D.F. , Annual R...
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