Method for fixing biological macro molecule pattern on polymer active surface by micro transfer technology

A technology of biological macromolecules and active surfaces, which is applied in the field of patterning and immobilizing biological macromolecules on polymer active surfaces with micro-transfer technology, can solve problems such as easy desorption and stability, and achieve high contrast, firmness and good application prospects. , the process is simple and flexible effect

Inactive Publication Date: 2003-06-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

But there are disadvantages of easy desorption and poor stability

Method used

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  • Method for fixing biological macro molecule pattern on polymer active surface by micro transfer technology
  • Method for fixing biological macro molecule pattern on polymer active surface by micro transfer technology
  • Method for fixing biological macro molecule pattern on polymer active surface by micro transfer technology

Examples

Experimental program
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Effect test

example 1

[0026] Example 1 The method for immobilizing biomacromolecules in a patterned polymer active surface with micro-transfer technology of the present invention comprises the following steps:

[0027] 1. Prepare the PCL film with free aldehyde groups on the surface: the PCL film is placed in the isopropanol solution of hexamethylenediamine with a concentration of 10% (wt), and it is aminated for 10 minutes at 37° C. Lots of free amine groups. Then put the PCL membrane rich in amine groups on the surface into 2% glutaraldehyde solution, and conduct a coupling reaction at 37° C. for 30 minutes to prepare the PCL membrane rich in free aldehyde groups on the surface. At this time, the film appears light red or orange red due to the aldylation reaction.

[0028] 2. Preparation of PDMS stamps with arrayed microwells on the surface: first prepare an original bottom mold with conventional "photolithography" technology, that is, on a clean 3×3cm 2 The surface of the glass slide was spin-...

example 2

[0035] This example is a comparative example with the present invention. The other steps were the same as in Example 1, but the existing reactive microcontact printing technology was used to pattern and immobilize albumin. That is, dip-coat the PBS buffer solution of FITC-BSA with a concentration of 1.0 mg / ml on the side of the PDMS stamp with microwells, and slowly dry it with nitrogen flow, and quickly transfer it on the aldehyde-formed PCL membrane. Pressure, time, temperature and subsequent flushing are all the same as in Example 1. CLSM observation found that the BSA pattern obtained by this method ( Figure 5 a) It is much darker than the BSA pattern obtained by the present invention, and its corresponding fluorescence intensity analysis shows that the pattern contrast is also much lower ( Figure 5 b). Example 3

example 3

[0036] The other steps are the same as in Example 1, but the PDMS stamp is left standing for 0.1 minute or 3 minutes in an atmospheric environment (25°C) with a relative humidity of 80-90%, and its influence on the pattern quality is observed. It was found that the former formed a ring-shaped pattern due to insufficient condensation water ( Figure 6 a), the latter causing pattern "contamination" due to excess condensation ( Figure 6 b). This shows that when using the microtransfer method to pattern and immobilize biomacromolecules, the amount of condensed water formed on the PDMS surface has a great influence on the quality of the final pattern, and too low or too high will lead to pattern defects.

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Abstract

The microwave transfer process of fixing biological macromolecule pattern to the active surface of polymer includes selectively "mounting" biological macromolecule to the micro wells of polydimethyl siloxane stamp; forming one condensated water layer on the surface; dissolving biological macromolecule in water of the micro well and of transverse diffused area and producing chemical reaction in solution; and fixing biological macromolecule in pattern on the active surface of the polymer. The process is simple, flexible and repeatable, and the micro pattern has high contrast and fastness. The cheap process of fixing biological macromolecule in pattern is especially suitable for the less reaction activity biological moecule/active surface system. The present invention has excellent application foreground in constructing biological device, tissue engineering, cell biological research, etc. based on cells.

Description

technical field [0001] The invention relates to a method for patterning and immobilizing biomacromolecules on the active surface of a polymer using micro-delivery technology. Specifically, the protein biomacromolecule solution "contained" in the microwells on the surface of the polydimethylsiloxane (PDMS) stamp surface array is transferred to the active surface of the polymer so that the reaction with the surface active groups can be carried out in a solution state. Therefore, it is more convenient to realize the patterned immobilization of protein biomacromolecules on the polymer surface. Background technique [0002] With the development of modern biotechnology, patterned immobilization of biological macromolecules has attracted extensive attention. Patterning and immobilizing biomacromolecules and cells on the surface of polymer materials has great practical value for specific biological detection, construction of biosensors based on living ce...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/00B41F3/00C08J3/24C12Q1/00G01N33/543G03F7/00
Inventor 高长有冯杰沈家骢
Owner ZHEJIANG UNIV
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