Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A real-time observation method of single cell of spore-forming bacteria

A real-time observation and spore technology, applied in the field of microorganisms, can solve problems such as rare bacteria research, and achieve the effect of simple operation and accurate observation data.

Active Publication Date: 2020-04-10
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although LSM is currently widely used in cell and molecular biology, tumor drug screening, and brain and neuroscience, it is rarely used in bacterial research.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A real-time observation method of single cell of spore-forming bacteria
  • A real-time observation method of single cell of spore-forming bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1: Cell Cycle Observation of Lysinibacillus varians GY32

[0018] Spread Lysinibacillus variants GY32 on an LB plate, culture it at 30°C for 7 days, then bake the plate at 105°C for 60 minutes, and examine the yield of spores under a microscope. Microscopic examination of colonies baked at a high temperature of 105°C showed no motile bacteria, only incomplete bacteria and spores with the outline of bacilli. It shows that high temperature treatment can kill all vegetative cells. Take the medium containing the lawn from the dry plate, add the medium containing the lawn to 1 mL LB liquid medium, break up the lawn, heat shock at 30°C for 30 minutes, and then immediately ice-bath for 15 minutes. Take 20 μL of the bacterial solution after ice bathing and add it to 2 mL of LB semi-solid medium containing 0.5% low-melting point agarose by mass fraction and mix evenly. Microscope real-time online observation. Microscopic examination shows that the real-time observatio...

Embodiment 2

[0019] Embodiment 2: Bacillus subtilis (Bacillus subtilis) 168 cell cycle observation

[0020] Bacillus subtilis (Bacillus subtilis) 168 was spread on an LB plate, cultured at 28°C for 10 days, and then the plate was baked at 80°C for 60 minutes, and the yield of spores was examined under a microscope. Microscopic examination of the colonies baked at 80°C showed no motile bacteria, only incomplete bacteria and spores with the outline of bacilli. It shows that high temperature treatment can kill all vegetative cells. Take the medium containing the lawn from the dry plate, add the medium containing the lawn to 1 mL LB liquid medium, break up the lawn, heat shock at 30°C for 30 minutes, and then immediately ice-bath for 15 minutes. Take 50 μL of the bacterial solution after ice bathing and add it to 2 mL of LB semi-solid medium containing 0.5% low-melting point agarose by mass fraction and mix evenly. Microscope real-time online observation. Microscopic examination shows that ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for real-time observation of single cells of spore generating bacteria. The method comprises the following steps: inoculating the spore generating bacteria to a culture medium for culture until spores are formed; then killing all nutrient cells to obtain the spores; then inoculating the spores to a semisolid culture medium; then transferring the spores together with the semisolid culture medium into a culture dish special for a laser confocal microscope; and observing single cells of the spore generating bacteria in real time by using the laser confocal microscope. According to the method disclosed by the invention, by collecting pure spores, optimizing spore germination conditions, optimizing the inoculation amount and controlling the agar concentration of the culture medium, a method for observing a growth cycle of a single cell of the spore generating bacteria online in real time is established. The real-time observation method disclosed by the invention does not need repeated sampling, thereby ensuring the accuracy of observation data and simple operation. The recording of the growth cycle of a single cell of the spore generating bacteria can be realized, thereby providing a new approach for online research of physiological characteristics of synchronous growth cells.

Description

Technical field: [0001] The invention belongs to the technical field of microbes, and in particular relates to a real-time observation method for a single cell of a spore-forming bacterium. Background technique: [0002] Spore bacteria widely exist in the atmosphere, water, soil, animals and plants and other environments. Spore-spore bacteria are one of the most commonly used model organisms in scientific research due to their ease of culture and their versatility. Research on its biological properties, spore formation and germination, and model systems for genetic differentiation has promoted the development of bacteriology. At the same time, its characteristics of spore formation, antibiotic production, flocculant production, protease and amylase production make it of special importance in the fields of biopesticides, pharmaceutical development, pollutant degradation and food processing. The development of appropriate research methods is of great benefit to strengthening...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/02
CPCC12Q1/02
Inventor 朱春节孙国萍许玫英郭俊
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com