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Anaerobic microorganism culture and real-time observation device

A technology for real-time observation of anaerobic microorganisms, applied in the field of anaerobic microorganism cultivation and real-time observation devices, can solve problems such as increased mortality, failure to achieve strict anaerobic levels, and difficulty in ensuring steady-state growth of anaerobic bacteria, and achieve improved experimental results. throughput, data quality assurance, and excellent airtightness

Pending Publication Date: 2021-06-04
SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the Desulfovibrio successfully cultivated by the institute is an anaerobic bacterium, many anaerobic bacteria can still survive under a certain oxygen concentration (2%-8%), but experiments have shown that it is difficult to survive in long-term culture. Ensure the steady-state growth (logarithmic growth) of anaerobic bacteria, and the mortality rate will be greatly improved; 2-10%), the physiological state is very different; the limitations of its device design can not reach the strict anaerobic level, and can not realize the further realization of liquid culture

Method used

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  • Anaerobic microorganism culture and real-time observation device
  • Anaerobic microorganism culture and real-time observation device
  • Anaerobic microorganism culture and real-time observation device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] This embodiment provides a kind of anaerobic microorganism culture and real-time observation device, such as figure 1 shown, which includes:

[0065] base 1 (eg figure 2 as shown), upper cover 2 (as image 3 shown), microfluidic chip, chip fixed plate 3 (such as Figure 4 Shown) and culture solution storage chamber (storage chamber body 4 and storage chamber loam cake 5 constitute, as Figure 6 and Figure 7 shown).

[0066] There are 8 evenly distributed and corresponding screw holes 17 and 26 on the edge of the contact surface where the base 1 and the upper cover 2 are fastened to each other, and the sealing and fastening between the base 1 and the upper cover 2 are realized by screws; the contact of the base 1 An annular groove 15 is formed on the edge of the surface, and the annular groove 15 is used for accommodating the rubber ring to further seal the base 1 and the upper cover 2 to ensure the airtightness of the device.

[0067]The base 1 is provided with ...

Embodiment 2

[0081] This embodiment provides a method for real-time observation and cultivation of anaerobic microorganisms, which uses the anaerobic microorganism cultivation and real-time observation device of Embodiment 1 to operate, including the following steps:

[0082] Prepare a microfluidic chip with a built-in anaerobic microorganism (the facultative anaerobic bacteria Escherichia coli used in this example) seed solution, the microfluidic chip consists of a glass slide and a chip structure sealed on the glass slide , the chip structure is communicated by the main channel with many sub-channels perpendicular to it, and the inlet port and the outlet port are connected with the main channel; the micro-control flow chip is placed in the third groove, and its inlet port passes through the pipeline and the culture solution storage chamber The outlet port is connected with the liquid outlet on the side wall of the upper cover; after the pipeline is connected, the upper cover and the botto...

Embodiment 3

[0086] In this implementation, an airtightness verification test is carried out for the anaerobic microorganism cultivation and real-time observation device in Example 1. When the gas is continuously fed, the whole device is placed in water without visible gas leakage, indicating that its airtightness is excellent.

[0087] For the verification of anaerobic atmosphere, Mitsubishi MGC oxygen indicator is used, and the experimental results are as follows Figure 10 shown. Depend on Figure 10 It can be seen intuitively that the oxygen concentration in the cultured gas environment is less than 0.1%, which is lower than the minimum detection limit.

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Abstract

The invention provides an anaerobic microorganism culture and real-time observation device. The device comprises a base, an upper cover, a micro-fluidic chip and a culture solution storage chamber; the base and the upper cover are mutually sealed and buckled, a first groove is used for accommodating the culture solution storage chamber, and a second groove is used for providing a culture space for anaerobic microorganisms; a third groove is used for accommodating a micro-fluidic chip, and a first window is formed in the horizontal groove surface of the third groove; a fourth groove is arranged opposite to the second groove, and a fifth groove is arranged opposite to the first groove; a second window is formed in the horizontal groove surface of the fourth groove; the first window and the second window are oppositely arranged; and a liquid outlet hole is formed in the culture solution storage chamber, and a gas inlet, a gas outlet and a liquid outlet are formed in the side wall of the upper cover. The device possesses excellent air tightness and can provide a stable anaerobic gas atmosphere for long-term stable liquid culture of anaerobic microorganisms, and the gas atmosphere is controllable; and the experimental requirements of high throughput, large-range multi-site sampling, high-magnification shooting and the like are met.

Description

technical field [0001] The invention belongs to the technical field of microorganism cultivation, and in particular relates to an anaerobic microorganism cultivation and real-time observation device. Background technique [0002] Long-term follow-up observation of a large number of individual bacterial cells has become an important means of microbial physiology research. Due to the advantages of flexible design, convenience and controllability, automation integration, real-time analysis, single-cell manipulation, saving samples and reagents, etc., microfluidic technology has been applied to a variety of microbial physiology studies. , Design related microfluidic chips. Regardless of how the chip is designed, its micro-scale characteristics enable researchers to use a special fixing device (CN112113901A) to fix it, and combine it with a time-lapse microscopy imaging system to achieve long-term autofocus shooting and data analysis. collection. [0003] Regarding time-lapse ...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12M1/34C12M1/04
CPCC12M23/16C12M23/22C12M23/48C12M23/38C12M29/20C12M41/36
Inventor 黄术强于跃马智鑫邓宇芳温慧李思宏
Owner SHENZHEN INST OF ADVANCED TECH CHINESE ACAD OF SCI
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