Cell sorter based on sacrificial-layer-free laser-induced forward transfer technology

Abstract

The invention relates to a cell sorter based on a sacrificial-layer-free laser-induced forward transfer technology, relates to the technical field of cell sorting, and solves the technical problems that fragments are easily generated and target cells are polluted during cell sorting in the prior art. The cell sorter sequentially comprises a femtosecond laser, a beam shaper, a sorting chip and a cell receiver in the light path direction, wherein the femtosecond laser and the beam shaper are respectively and electrically connected with the control system. According to the cell sorter based on the sacrificial-layer-free laser-induced forward transfer technology, target cells sorted based on the sacrificial-layer-free laser-induced forward transfer technology do not contain sacrificial-layer fragment pollution, and no pollution interference is caused to follow-up research and treatment of the obtained target cells.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of sorting cells, in particular to a cell sorter based on laser induce forward transfer technology without sacrifice layer. BACKGROUND

[0002] Laser induce forward transfer (LIFT) technology is concerned because it can accurately sort cells. The specific process is as follows: a suspension containing target cells is dispersed on the surface of a sorting chip with a sacrificial layer (usually a thin layer of metal); a pulsed laser induces a micro-explosion of the sacrificial layer at the target cell location, pushing the target cell away from the sorting chip and into a target cell receiving device, i.e. completing the sorting of the target cell. The sorted target cells can be used for scientific research. However, after the micro-explosion of the sacrificial layer, many fragments are generated and transferred into the target cell receiving device along with the target cells, which can contaminate the target cells and even change the biological properties of the sorted target cells, interfering with subsequent scientific research on the target cells. SUMMARY

[0003] The present application solves the technical problem of the prior art that sorting cells can easily produce fragments and contaminate target cells, and provides a cell sorter based on laser induce forward transfer technology without a sacrificial layer.

[0004] To solve the above technical problems, the technical solution of the present application is as follows:

[0005] A cell sorter based on laser induce forward transfer technology without a sacrificial layer, in the direction of the optical path, includes in sequence: a femtosecond laser, a beam shaper, a sorting chip and a cell receiver; wherein the femtosecond laser and the beam shaper are also electrically connected to a control system;

[0006] The femtosecond laser is used to output femtosecond laser into the beam shaper;

[0007] The beam shaper is used to modulate the femtosecond laser output by the femtosecond laser to generate a single beam or multiple beams of femtosecond laser;

[0008] The sorting chip includes: a sacrificial layer-free chip and a micro-well membrane, the micro-well membrane is arranged below the sacrificial layer-free chip, the micro-well membrane contains a plurality of micro-wells; the target cells are located in the micro-wells; the sacrificial layer-free chip is used to transmit the laser beam output by the femtosecond laser; the micro-well membrane is used to adjust the size of the micro-wells to adjust the number of target cells loaded in a single micro-well, and further adjust the number of target cells sorted in a single sorting process;

[0009] The control system is used to control the number, number and morphology of the femtosecond laser pulses output by the femtosecond laser and the beam shaper.

[0010] In the above technical solution, the pulse width of the femtosecond laser ranges from 1 fs to 999 fs.

[0011] In the above technical solution, the pulse width of the femtosecond laser ranges from 1 fs to 100 fs.

[0012] In the above technical solution, the single pulse energy of the femtosecond laser ranges from 1 pJ to 1 μJ.

[0013] In the above technical solution, the single pulse energy of the femtosecond laser ranges from 10 nJ to 500 nJ.

[0014] In the above technical solution, the wavelength of the femtosecond laser ranges from 350 nm to 2000 nm.

[0015] In the above technical solution, the wavelength of the femtosecond laser ranges from 400 nm to 800 nm.

[0016] In the above technical solution, the beam shaper modulates the shape and energy of the femtosecond laser beam; when multiple femtosecond laser beams are generated, the beam shaper is used to control the spatial position relationship between the generated multiple femtosecond laser beams, the shape, energy, and number of output pulses per unit time of each femtosecond laser beam; the beam shaper modulates the focusing of the femtosecond laser between the sacrificial layer-free chip and the target cell, inside the micro-trap, at this position, the nonlinear breakdown of the solution generates a bubble; and further controls each femtosecond laser focused between the sacrificial layer-free chip and the target cell, the size of the bubble induced and generated and its movement direction.

[0017] In the above technical solution, the beam shaper is a mirror group, a spatial light modulator, or a digital micro-mirror.

[0018] In the above technical solution, the micro-trap film is a single-layer micro-trap film or a stacked micro-trap film.

[0019] The present application has the following beneficial effects:

[0020] The cell sorter based on the sacrificial layer-free laser-induced forward transfer technology of the present application has the following beneficial effects: BRIEF DESCRIPTION OF DRAWINGS

[0021] The present application will be further described in detail below in combination with the drawings and specific embodiments.

[0022] Figure 1 The structure of the cell sorter based on the sacrificial layer-free laser-induced forward transfer technology of the present application is shown in the figure.

[0023] The reference signs in the figure are as follows:

[0024] 101-femtosecond laser; 102-beam shaper; 103-sorting chip; 104-cell receiver; 105-control system;

[0025] 1031-sacrificial layer-free chip; 1032-micro-well membrane; 1033-bubble; 1034-target cell; 1035-micro-well. DETAILED DESCRIPTION

[0026] The inventive idea of the present application is:

[0027] Compared with the conventional LIFT technology in the prior art for sorting cells by means of micro-explosion of a sacrificial layer, the femtosecond laser can nonlinearly break the solution to generate a bubble and a jet, and further push the adjacent cells to move, i.e. the target cells can be pushed to move without a sacrificial layer.

[0028] Specifically, a suspension containing target cells is dispersed on the surface of a sorting chip without a sacrificial layer; the femtosecond laser is focused on the liquid layer between the target cells and the sorting chip, the solution is nonlinearly broken to generate a bubble and a jet, and further the target cells are pushed away from the sorting chip and into a target cell receiving device. In this process, there is no sacrificial layer fragment with the target cells into the target cell receiving device, so that the scientific research on the obtained target cells will not be interfered by the foreign sacrificial layer fragment impurities. The cell sorter based on the sacrificial layer-free laser-induced forward transfer technology of the present application is realized based on femtosecond laser technology.

[0029] The present application will be described in detail below with reference to the accompanying drawings.

[0030] Referring to Figure 1 The cell sorter based on the sacrificial layer-free laser-induced forward transfer technology of the present application sequentially comprises, in the direction of the optical path: a femtosecond laser 101, a beam shaper 102, a sorting chip 103 and a cell receiver 104; wherein the femtosecond laser 101 and the beam shaper 102 are further electrically connected with a control system 105;

[0031] The femtosecond laser 101 is used to output femtosecond laser into the beam shaper 102; the pulse width of the femtosecond laser 101 ranges from 1 fs to 999 fs, preferably from 1 fs to 100 fs; the single pulse energy ranges from 1 pJ to 1 μJ, preferably from 10 nJ to 500 nJ; and the wavelength ranges from 350 nm to 2000 nm, preferably from 400 nm to 800 nm.

[0032] The beam shaper 102 is used to modulate the shape and energy of the femtosecond laser beam, to generate a single beam of femtosecond laser, multiple beams of femtosecond laser, and to control the spatial position relationship between the multiple beams of femtosecond laser, the shape of each beam of femtosecond laser, the energy, and the number of output pulses per unit time when multiple beams of femtosecond laser are generated. The beam shaper 102 modulates the focusing of the femtosecond laser between the sacrificial layer-free chip 1031 and the target cell 1034, inside the micro-trap 1035, at which position the bubble 1033 is generated by the nonlinear breakdown of the solution. The beam shaper 102 further controls each beam of femtosecond laser focused between the sacrificial layer-free chip 1031 and the target cell 1034, and the size and movement direction of the bubble 1033 induced and generated. The beam shaper 102 is specifically a mirror group, a spatial light modulator, or a digital micro-mirror.

[0033] The sorting chip 103 includes a sacrificial layer-free chip 1031 and a micro-trap membrane 1032 arranged below the sacrificial layer-free chip 1031, and the micro-trap membrane 1032 contains multiple micro-traps 1035. The target cell 1034 is located in the micro-trap 1035. The sacrificial layer-free chip 1031 is used to transmit the laser beam output by the femtosecond laser 101. The micro-trap membrane 1032 is a single-layer micro-trap membrane or a laminated micro-trap membrane. The micro-trap membrane 1032 is used to adjust the size of the micro-trap 1035, to adjust the number of target cells loaded in the single micro-trap 1035, and to further adjust the number of target cells 1034 sorted in a single sorting process. When the beam shaper 102 modulates the focusing of the femtosecond laser between the sacrificial layer-free chip 1031 and the target cell 1034, inside the micro-trap 1035, the bubble 1033 is generated by the nonlinear breakdown of the solution at this position.

[0034] The control system 105 is electrically connected to the femtosecond laser 101 and the beam shaper 102, respectively, to control the operation of the femtosecond laser 101 and the beam shaper 102. The control system 105 can control the number of femtosecond laser pulses, the number and shape of the femtosecond laser beam output by the femtosecond laser 101 and the beam shaper 102.

[0035] In summary, the cell sorter based on the sacrificial layer-free laser-induced forward transfer technology according to the present application sorts the target cells without the contamination of sacrificial layer fragments, and does not interfere with the subsequent research and processing of the obtained target cells.

[0036] Obviously, the above embodiments are only examples for clear illustration, and are not intended to limit the embodiments. Based on the above description, other different forms of changes or variations can be made by those skilled in the art. Here, all the embodiments cannot be exhaustively listed. The changes or variations derived therefrom are still within the protection scope of the present application.

Claims

1. A cell sorter based on a sacrifice-free laser-induced forward transfer technique, characterized in that, The optical path direction comprises, in sequence: a femtosecond laser (101), a beam shaper (102), a sorting chip (103) and a cell receiver (104); wherein the femtosecond laser (101) and the beam shaper (102) are also electrically connected with a control system (105) respectively; The femtosecond laser (101) is used to output femtosecond laser into the beam shaper (102); The beam shaper (102) is used to modulate the femtosecond laser output by the femtosecond laser (101) to generate single or multiple beams of femtosecond laser; The sorting chip (103) comprises: a sacrificial layer-free chip (1031) and a micro-well membrane (1032), the micro-well membrane (1032) is arranged below the sacrificial layer-free chip (1031), and the micro-well membrane (1032) contains a plurality of micro-wells (1035); target cells (1034) are located in the micro-wells (1035); the sacrificial layer-free chip (1031) is used to transmit the laser beam output by the femtosecond laser (101); the micro-well membrane (1032) is used to adjust the size of the micro-well (1035) to adjust the number of target cells loaded in the single micro-well (1035), and then adjust the number of target cells (1034) sorted out in a single sorting process; The control system (105) is used to control the number of femtosecond laser pulses, the number and the shape of the femtosecond laser beam output by the femtosecond laser (101) and the beam shaper (102).

2. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technology according to claim 1, characterized in that, The pulse width of the femtosecond laser (101) ranges from 1 fs to 999 fs.

3. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 2, characterized in that The pulse width of the femtosecond laser (101) ranges from 1 fs to 100 fs.

4. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 1, characterized in that, The single pulse energy of the femtosecond laser (101) ranges from 1 pJ to 1 μJ.

5. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 4, characterized in that The single pulse energy of the femtosecond laser (101) ranges from 10 nJ to 500 nJ.

6. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 1, characterized in that, The wavelength of the femtosecond laser (101) ranges from 350 nm to 2000 nm.

7. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 6, characterized in that The wavelength of the femtosecond laser (101) ranges from 400 nm to 800 nm.

8. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 1, characterized in that, The beam shaper (102) modulates the shape and energy of the femtosecond laser beam; when multiple beams of femtosecond laser are generated, the beam shaper (102) is used to control the spatial position relationship between the generated multiple beams of femtosecond laser, the shape, energy and number of output pulses per unit time of each beam of femtosecond laser beam; the beam shaper (102) modulates the focusing of the femtosecond laser between the sacrificial layer-free chip (1031) and the target cells (1034), inside the micro-well (1035), at this position, the bubble (1033) is generated by the nonlinear breakdown of the solution; and then control each beam of femtosecond laser focused between the sacrificial layer-free chip (1031) and the target cells (1034), the size and movement direction of the bubble (1033) induced and generated.

9. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 1, characterized in that, The beam shaper (102) is a mirror group, a spatial light modulator or a digital micro-mirror.

10. The cell sorter based on the sacrifice-free layerless laser-induced forward transfer technique according to claim 1, characterized in that, The micro-well membrane (1032) is a single-layer micro-well membrane or a stacked micro-well membrane.

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