Method for manufacturing micro-hemisphere array plate, microfluidic device comprising micro-hemisphere array plate, and method for culturing cell aggregate using same
An array plate and aggregate technology, applied in the field of preparation of micro-hemisphere array plates, can solve the problems of inability to separate, cell separation, etc.
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[0034] Specifically, the method for preparing the micro-hemispherical array plate of the present invention includes:
[0035] 1) Step, attach a photosensitive photoresist material on the silicon substrate;
[0036] 2) Step, adjusting the height of the photosensitive photoresist material by spin coating;
[0037] 3) Step, etching the photoresist material to a hemispherical shape by overcure;
[0038] 4) step, after the step 3), vapor-deposit the first metal layer on the etched surface;
[0039] 5) Step, after the first metal layer is vapor-deposited, a second metal layer is vapor-deposited thereon;
[0040] 6) Step, forming a core layer on the second metal layer;
[0041] 7) step, after the step 6), planarize the upper surface of the core layer;
[0042] 8) step, after the step 7), separating the core layer;
[0043] 9) step, using the separated core layer as a template to perform injection molding to obtain a micro hemispherical array plate; and
[0044] 10) Step of imparting hydrophilicity ...
Embodiment 1
[0074] Example 1: Fabrication of micro hemisphere array plate and cultivation of cell aggregates
[0075]
[0076] A photosensitive photoresist material is used in order to embody a micro-hemispherical pattern with a size of 500 um. In this case, the photosensitive photoresist material used can be Negative and Positive. In the present invention, the Negative form is used.
[0077] The photosensitive photoresist material is spin-coated in the 300um area to adjust the height of the micro-hemispheres, and the coated photoresist material can be visualized as the micro-hemispheres through over-vulcanization at a temperature of 150°C.
[0078] The first seed metal layer is placed on the micro-hemispherical array pattern formed on the silicon substrate using thin film evaporation equipment. At this time, titanium is used as the seed metal, and its height is The second metal layer uses nickel (Nickel), its height is The first and second metal thin film evaporation equipment used E-beam evap...
Embodiment 2
[0091] Example 2: Cultivation of cell aggregates using microfluidic devices including micro-hemisphere array plates
[0092]
[0093] A microfluidic device including the micro hemispherical array plate prepared by the method of Example 1 was fabricated. This microfluidic device can be divided into a sample injection part, a sample mixing part, and a cell assembly forming part including a micro-hemisphere array plate. the following Figure 5 It is a cross-sectional view. And, the following Image 6 This is a picture of a microfluidic device including a micro hemispherical array plate prepared in the manner shown above.
[0094] 1) Isolation of human liver cells
[0095] The liver tissues cut from patients undergoing partial hepatectomy are separated from human liver cells using the conventional collagenase-two-step method. To briefly explain, for the separated liver tissue, firstly remove blood by perfusion with EGTA, and then use type 2 collagenase solution (type2collagenase) for ...
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Abstract
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