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High-throughput automatic sorting and receiving system for microorganism single cells

A receiving system and single-cell technology, applied in the direction of material analysis, measuring devices, and analysis materials through optical means, can solve the problem of less cell information, achieve high processing throughput, low hardware cost, automatic sorting and receiving Effect

Inactive Publication Date: 2017-12-19
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although flow cytometry can analyze single cells at high speed and high throughput, this also results in less cell information that can be obtained in a very short detection time, and it is often necessary to obtain more abundant and Accurate information without visualizing individual cells for long-term observation and detection by other modalities such as Raman spectroscopy

Method used

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  • High-throughput automatic sorting and receiving system for microorganism single cells
  • High-throughput automatic sorting and receiving system for microorganism single cells
  • High-throughput automatic sorting and receiving system for microorganism single cells

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1: Microbial single cell sorting based on fluorescence imaging

[0041] figure 1 Shown is a schematic diagram of a high-throughput single-cell automatic sorting and receiving system. It can be seen from the figure that the present invention provides a high-throughput single-cell automatic sorting and receiving system, including a microscopic imaging module 1, a pulsed laser module 2, and a cell separation module. 3. The cell receiving module 4 and the control terminal 5, the connection relationship of the above components is as follows:

[0042]The microscopic imaging module 1 is based on an upright microscope, and includes a CCD 1-1 for imaging and a plurality of focusing objective lenses 1-2. The pulsed laser module 2 is coupled with the microscopic imaging module 1 through an optical path 2-1. Cells The separation module 3 is placed in the microscopic imaging module 1. The cell separation module 3 includes the first automatic displacement loading platform an...

Embodiment 2

[0050] Embodiment 2: Microbial single cell sorting based on Raman spectroscopy

[0051] The following is an embodiment of microbial single cell sorting based on Raman spectroscopy: wherein:

[0052] The microscopic imaging module 1 includes a commercial upright Raman microscope system (model BX-41, purchased from Olympus);

[0053] Pulse laser module 2 is a pulse laser with a wavelength of 532nm (model PULSELAS-P-532-300, purchased from ALPHALAS company), the laser pulse width is 1nm, the maximum single pulse energy is 10μJ, and the pulse frequency is: 5-16.6kHz;

[0054] The cell separation module 3 includes a first automatic displacement loading platform and a laser ejection substrate, the base layer material of the laser ejection substrate is silicate glass; the coating film material is ITO;

[0055] The cell receiving module 4 includes a second automatic displacement loading platform and a 384-well plate.

[0056] The method for using the above-mentioned high-throughput ...

Embodiment 3

[0058] Example 3: Single cell sorting coupled with single cell genome manipulation

[0059] In order to further verify that the single cells sorted by the above-mentioned high-throughput single cell automatic sorting and receiving system are the desired cells, the single cells obtained by sorting are subjected to genome manipulation. The operation process is as follows:

[0060] Escherichia coli single cells in Example 1 were ejected and sorted into 9 receiving microwells, alkaline lysate was added to lyse the cells, and then transferred to centrifuge tubes and nucleic acid amplification mixed reagents and amplification enzymes were added, using The MDA amplification method amplifies the single-cell genome, and performs PCR verification on the product, and obtains as follows: Figure 4 In the gel electrophoresis images shown, #1 to #9 in the figure are the results of single cells obtained by ejection sorting, and N1 and N2 are negative controls that do not contain any cells. ...

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Abstract

The invention relates to a high-throughput automatic sorting and receiving system for microorganism single cells. The system comprises a microscope imaging module, a cell separation module, a pulse laser module, a cell receiving module and a control terminal. An sorter for the microorganism single cells can automatically identify the microorganism single cells, and can couple optical images, fluorescence images, Raman spectrum and various other observation modes to realize the rapid and automatic sorting and receiving of the microorganism single cells at a low hardware cost and a high processing throughput.

Description

technical field [0001] The invention relates to the field of microbial single-cell sorting equipment, and more specifically relates to an instrument system capable of coupling multi-modal cell measurement and simultaneously realizing high-throughput, fast, automatic sorting and receiving of microbial single cells. Background technique [0002] Microorganisms are an important resource for species on the earth, and their biomass accounts for about half of the total global biomass. Through cell engineering and genetic engineering transformation of microorganisms, humans have produced enormous value in the fields of industry, agriculture, medicine, and environmental protection. However, at present, human beings' understanding and utilization of the earth's microorganisms are still very limited, and more than 90% of the microbial species still belong to the "dark matter (dark matter)" of the biological world because they cannot be cultivated artificially. Currently, there are ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/01G01N21/85G01N35/00
Inventor 勾洪磊籍月彤徐健
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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