Sorter and sorting method for separating cells and particles
A particle separation and sorting technology, applied in the biological field, can solve the problems of not being able to expand the single-cell level, not being able to sort cells/particles, etc.
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Embodiment 1
[0066] Example 1 Construction of Raman Flow Sorter for Cell and Particle Separation
[0067] 1) Raman spectrum acquisition and analysis system
[0068] The Raman flow sorting instrument used for the separation of cells and particles constructed in this example, the Raman spectrometer in the Raman spectrum acquisition and analysis system uses a confocal micro Raman spectrometer (PrincetonInstrument, US), and the excitation wavelength is 514.5nm (Coherent, Innova 90-5 Ar-ion). A Leica DM-IRB microscope with a 63x objective lens (Leica, HCXPL APO) was used for nano-microscale microscopy. Raman spectrum signal acquisition and analysis software uses Labs pec 5.0.
[0069] 2) Microfluidic control system
[0070] In the Raman flow sorter microfluidic control system for cell and particle separation constructed in this example, the inner diameter of the sample injection channel of the microfluidic device is 100 μm, and the length is 20 mm; the inner diameter of the protective liquid...
Embodiment 2
[0073] Example 2 Sorting of isotope-labeled cells by a Raman flow sorter for cell and particle separation
[0074] (1) Prepare target cell / particle samples and prepare sorting protection solution
[0075] Use liquid mineral medium as cell culture medium, 1 liter of liquid mineral medium contains 2.5g Na 2 HPO 4 , 2.5g KH 2 PO 4 , 1.0g NH 4 Cl, 0.1g MgSO 4 ·7H 2 O, 10 μL saturated CaCl 2 solution, 10 μL saturated FeSO 4 solution and 1mL Bauchop & Elsden. Among them, 1L of Bauchop& Elsden solution contains 10.75g MgSO4, 4.5g FeSO 4 ·7H 2 O, 2.0g CaCO 3 , 1.44gZnSO 4 ·7H 2 O, 1.12g MnSO 4 4H 2 O, 0.25g CuSO 4 ·5H 2 O, 0.28g CoSO 4 ·7H 2 O, 0.06g H 3 BO 3 and 51.3 mL of saturated HCl solution.
[0076] Add 100 μL of seawater sample to 10 mL of liquid mineral medium, and add 1.70 mg of carbon-13-labeled NaHCO at the same time 3 As the only carbon source, its concentration is 2mM. Shake culture at 30°C for 14 days to obtain a cell solution to be sorted.
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