Hydrogel particles with tunable optical properties and methods for using same
A technology of hydrogel particles and optical properties, applied in biochemical equipment and methods, biological particle analysis, particle and sedimentation analysis, etc., can solve problems such as batch differences, expensive and laborious
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Embodiment 1
[4466] Example 1: Generation of Hydrogel Particles
[4467] Photomasks for UV lithography were obtained from CADart Services Inc. and designed using AutoCad (AutoDesk, Inc.). Photocrosslink SU-8 photolithographic resist (Microchem, Inc.) on a 4" silicon wafer using a calibrated UV light source (OAI, Inc.) to create a mask for microfluidic device fabrication. PDMS (Polydimethylene Silicone based siloxane, Sigma Aldrich, Inc.) was prepared and shaped using soft lithography and standard published methods for microfluidic device fabrication (see, McDonald JC, et al., 2000, Electrophoresis 21:27-40).
[4468]Droplets are formed using a flow-focusing geometry in which two oil channels converge the central flow of the aqueous monomer solution, thus breaking the droplets in the water-in-oil emulsion. For droplet formation, fluorocarbon oil (Novec 75003M, Inc.) was used as the external continuous phase liquid. To stabilize the droplets before polymerization, a surfactant was added ...
Embodiment 2
[4472] Example 2: Generation and imaging of 12 11m hydrogel particles
[4473] Water containing 5% acrylamide, 0.25% bisacrylamide, 0.05% allylamine, and 0.1% ammonium persulfate was flowed through the center channel and converged by oil containing 0.1% TEMED through a 10 micron nozzle to produce 10 μm hydrogel particles, such as Figure 3A shown. After polymerization, the particles were washed in water, as Figure 3B shown and conjugated to the relevant dyes. Fluorescent hydrogel particles were visualized with a fluorescence microscope, as Figure 3C shown.
Embodiment 3
[4474] Example 3: Multidimensional Adjustment of Optical Properties of Hydrogel Particles
[4475] As described in Figure 4, the hydrogel particles are tuned in multiple dimensions to match specific cell types other than polystyrene beads. Using optical parameters such as FSC and SSC ( Figure 4A ) or a combination of secondary markers to deconvolute the cells. Hydrogel particles are tuned to match specific cell types SSC and FSC other than polystyrene beads (brown), which are limited in size (FSC) and side scatter ( Figure 4B ). The hydrogel particles are further functionalized with stoichiometrically tuned specific chemical side groups and secondary markers, allowing precise cell type matching without compromising biological noise and still behaving like fixed cell lines ( Figure 4C ).
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