An electrospray ionization device and its application

A technology of electrospray ionization and conductive clips, applied in the direction of measuring devices, ion sources/guns, circuits, etc., can solve problems such as opacity, limited applications, poor biocompatibility, etc., achieve high biocompatibility, improve The effect of precision and repeatability

Inactive Publication Date: 2018-05-18
CHINA UNIV OF GEOSCIENCES (BEIJING)
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AI-Extracted Technical Summary

Problems solved by technology

However, the poor biocompatibility of these paper genes makes it difficult for cells to attach to them, and their opacit...
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Abstract

The invention relates to an electrospray ionization device, comprising: a glass substrate having at least one tip, on which a plurality of microchannels are arranged; a conductive clip for applying a high voltage and clamping the glass substrate; and for placing the glass-based chamber. The invention also relates to the application of the electrospray ionization device in cell-based drug analysis. The glass substrate provides high biocompatibility, which can be used for qualitative and quantitative analysis of drugs in cells, and can also be used for biological analysis of cells after drug action.

Application Domain

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  • An electrospray ionization device and its application
  • An electrospray ionization device and its application
  • An electrospray ionization device and its application

Examples

  • Experimental program(5)

Example Embodiment

[0048] Example 1 Manufacturing method and structure of electrospray ionization device
[0049] (Production of glass substrate)
[0050] Cut a cover glass with a thickness of 20μm with a glass knife into an isosceles triangle with a waist length of 0.8cm and an apex angle of 45°. After washing and drying, spin a layer on the surface with a homogenizer at 1000rpm The SU-82015 negative photoresist with a thickness of 30μm is baked at 65°C for 20min, and a mask with a microchannel pattern (the width of the microchannel is 2mm, the number is 3) is covered on the glass slide, After ultraviolet exposure, development, and nitrogen drying, a glass substrate with 3 microchannels with a width of 2 mm and a depth of 30 μm is obtained.
[0051] (Production of PDMS chamber)
[0052] The PDMS prepolymer and the initiator are mixed at a mass ratio of 10:1 and then poured on the silicon wafer. The silicon wafer is sealed in advance with a wide tape to prevent the PDMS prepolymer from leaking. After removing bubbles, the PDMS prepolymer was placed at 75° C. for 2 hours to carry out the polymerization reaction. The polymerized PDMS sheet was carefully peeled off, and cut into a PDMS sheet of 2cm×2cm×5mm. Use the same method to make another PDMS sheet of the same size, and dig out an isosceles triangle cavity (the waist length of the isosceles triangle cavity is 1.2 cm, and the vertex angle is 45°). The PDMS sheet with the isosceles triangle cavity dug and the PDMS sheet without the cavity are laminated together to form a PDMS chamber that can accommodate the glass substrate, and the glass substrate is placed in it.
[0053] Conductive clip: prepare a conductive metal clip for use.

Example Embodiment

[0054] Example 2 Co-cultivation of cells in microchannels
[0055] 1) Digest the human liver tumor cells HepG2, human cloned colon adenocarcinoma cell Caco-2 and human breast cancer cell MCF-7 cells that are overgrown in a 60mm petri dish, and dilute them with culture medium to 10 6 mL -1 Cell suspension. Take 10 μL of the cell suspension of the three kinds of cells and add them to the three microchannels on the glass substrate in the PDMS chamber (e.g. figure 2 (Shown), after the cells adhere to the wall, add some new medium, put the glass substrate placed in the PDMS chamber into the cell incubator, and culture overnight.
[0056] 2) Take out the glass substrate from the incubator, stain HepG2, Caco-2 and MCF-7 cells with three different fluorescent dyes, Cell TrackerTM Green CMFDA, CellTrackerTM Deep Red and Cell TrackerTM Violet, and place them in a confocal laser Observe under the microscope, the results are as image 3 Shown. It can be seen that due to the surface tension of the microchannel, the three kinds of cells grow in their respective microchannels, and the cells will not mix with each other.
[0057] 3) In the same way as in step 1), culture the three types of HepG2, Caco-2 and MCF-7 cells overnight, and change the medium every day for standby. The cell survival rate was tested daily with a deadly kit (Calcein-AM/EthD-1). The result is Figure 4 As shown, the cell activity remained normal during the first three days of culture, and the cell survival rate still reached over 85% on the third day.

Example Embodiment

[0058] Example 3 Qualitative mass spectrometry detection of drug absorption in cells
[0059] Co-culture HepG2, Caco-2 and MCF-7 cells in the same manner as step 1) of Example 2. After culturing overnight, remove the glass substrate placed in the PDMS chamber from the incubator, and add cyclophosphamide dropwise The solution (hereinafter referred to as CPA, the concentration is 10 mmol/L, that is, 10 mM, the dosage is 10 μL), and then put it back into the incubator for 24 hours.
[0060] A glass substrate with MCF-7 cells (ie, monocultured MCF-7 cells) cultured in all three microchannels was used as a control sample, and the culture mode and drug action mode were the same as the co-cultured MCF-7 cells.
[0061] Remove the glass substrate placed in the PDMS chamber from the cell incubator, clamp the glass substrate, and rinse with distilled water three times to remove the complex matrix remaining on the surface of the glass slide. After the glass substrate is naturally dried, place it in front of the Shimadzu LCMS-2010A mass spectrometer with a conductive clip, so that the tip of the glass substrate (at an angle of 45°) faces the inlet of the mass spectrometer, and apply the conductive clip to the glass substrate High voltage (~4.5kV), while adding isopropanol as an organic solvent into the microchannel where the MCF-7 cells are located, an electrospray is generated immediately, and the spray ions are collected by a mass spectrometer and scanned and analyzed. The mass-to-charge ratio (m/z) of the scan ranges from 100 to 500, and the detection is performed in the positive ion mode. The control sample was also scanned and analyzed in the above-mentioned manner.
[0062] Such as Figure 5 with 6 As shown, there is [CPA+H] in all mass spectra + Peak (m/z is 262), [CPA+Na] + Peak (m/z is 284) and [CPA+K] + Peak (m/z is 300). These typical mass spectrum peaks indicate that CPA is absorbed in both monoculture and co-cultured MCF-7 cells.
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PUM

PropertyMeasurementUnit
Width1.0 ~ 3.0mm
Depth10.0 ~ 50.0µm
Side length<= 1.0cm
tensileMPa
Particle sizePa
strength10

Description & Claims & Application Information

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