Improved flow cytometer based on optical fiber integrated micro-fluidic chip

[0034]Example 1:

[0035]Such asfigure 1 Shown is a system diagram of an improved flow cytometer based on a fiber integrated microfluidic chip. It is composed of five modules: optical fiber integrated microfluidic chip 1, light source 2, fluid control system 6, photoelectric detection system 7, and waste liquid collection system 9. In the system, the fluid control system 6 controls the cell sample and the sheath fluid to form a single cell flow, and flows into the cell flow conduit 10 of the microfluidic chip 1, and the cell flow conduit 10 is connected to the hollow annular core optical fiber 4. The hollow ring-core fiber 4 is connected to the optical fiber 3 to introduce the excitation light of the light source 2. The connection between the hollow ring-core fiber 4 and the optical fiber 3 can be in the form of a coupler, such asFigure 8 As shown in (a), it can also be in the form of core alignment, such asFigure 8 (b) Shown. The hollow ring-core optical fiber 4 uses its own hollow part 4 to 5 as a micro flow channel to allow cell flow to pass. The output end of the hollow ring core optical fiber 4 is made into a frustum structure by fiber grinding technology, which is used for focusing and irradiating the excitation light. The cells 11 in the cell flow are due to the self-focusing effect of the beam at the output end of the hollow ring core optical fiber 4 , Only one can pass at a time at the autofocus, as shown in Figure 8. After the cells 11 in the cell stream are irradiated by the excitation light source, they are collected and processed by the waste liquid collection system 9. The scattered light generated by the irradiation is collected by scattered light collection fibers 8-1, 8-2, and transmitted to the photoelectric detection system 7-1, 7-2 for corresponding detection and analysis.

[0036]PE-TxRed was selected as the fluorescent marker of the cell sample, and the excitation light source wavelength was 488nm. After the PE-TxRed attached to the cells in the single-cell flow is irradiated by the excitation light source, the generated scattered excitation light has a wavelength of 620nm. The scattered light is collected by the scattered light collection fibers 8-1 and 8-2 and transmitted to the photoelectric detection system 7 respectively. -1, 7-2. Take the photoelectric detection system 7-2 as an example to illustrate the scattering and detection process of scattered light.

[0037]The scattered light collecting fiber 8-2 collects scattered light of two wavelengths, namely: the excitation light of the light source with a wavelength of 488nm and the excited light of 620nm. The scattered light collection optical fiber 8-2 transmits the collected scattered light to the photodetection system 7-2. The photodetection system 7-2 contains a spectroscopic module 12 and a photodetection module 13. The beam splitting surfaces of the beam splitters 12-1 and 12-2 of the beam splitting module 12 are respectively plated with 500nm long-wave pass dichroic mirrors and short-wave pass dichroic mirrors. Correspondingly, the single photon detectors 13-1 and 13-2 in the photoelectric detection module 13 are used to detect scattered light with a wavelength of 488 nm and scattered light with a wavelength of 620 nm, respectively.

[0038]Example 2:

[0039]FITC and APC-Cy7 were selected as the markers of the cells in the single-cell flow. The excitation light wavelengths were 488nm and 635nm, and the excited light wavelengths were 530nm and 780nm.

[0040]The light sources 2-1 and 2-2 are laser light sources of 488 nm and 635 nm, respectively, which are combined by the wavelength division multiplexer 14 and then transmitted to the hollow ring core fiber 4 via the optical fiber 3. The cells 11 in the cell flow generate scattered light after being irradiated, and the scattered light is collected by the scattered light collecting fibers 8-1, 8-2, and transmitted to the photoelectric detection systems 7-1, 7-2, respectively. The scattered light contains a total of 4 wavelengths of light, the excitation light with wavelengths of 488nm and 635nm, and the excited light with wavelengths of 530nm and 780nm. Take the photoelectric detection system 7-2 as an example to illustrate the spectroscopy and detection of scattered light.

[0041]There are four wavelengths of scattered light entering the photoelectric detection system. The light splitting surface of the light splitting interface 12-1 is coated with a short-wavelength pass dichroic mirror with a wavelength of 700nm. The scattered light with a wavelength of 780 enters the photoelectric detection module 13 first. The photon detector 13-1 carries out the corresponding detection processing. The spectroscopic surface of the spectroscopic interface 12-2 is coated with a short-wavelength pass dichroic mirror with a spectroscopic wavelength of 600nm, and the wavelength of 635nm enters the photodetector module 13 through the spectroscopic interface 12-2. , The single-photon detector 13-2 carries out corresponding detection processing, and the spectroscopic surface of the spectroscopic interface 12-3 is coated with a short-wavelength pass dichroic mirror with a wavelength of 500nm, and the scattered light with a wavelength of 530nm enters through the spectroscopic interface 12-3 The photodetection module 13 is processed by the single-photon detector 13-3. Finally, the light-splitting interface of the light-splitting interface 12-4 is coated with a long-wavelength pass dichroic mirror with a wavelength of 500nm, and the scattered light with a wavelength of 488nm passes through the light-splitting interface 12-4 enters the photodetection module 13, and the single-photon detector 13-4 performs corresponding detection processing.