Preparation method of TEM (transmission electron microscope) ultrathin section samples of phosphorus-accumulating bacteria

Abstract

The invention relates to a preparation method of TEM (transmission electron microscope) ultrathin section samples of phosphorus-accumulating bacteria, which comprises the following steps: after a sample is taken, carrying out centrifugal cleaning on the sample, putting the phosphorus-accumulating bacteria PAOs sample into a glutaraldehyde solution with a pH value being 7.4 and a volume percentage concentration of 2.5% to fix; dyeing the phosphorus-accumulating bacteria sample by using a Pb(NO3)2 solution with a mass percentage concentration of 2-10%; carrying out dehydration by using a gradient ethanol-acetone series dehydration method; and carrying out resin impregnation and embedding, and sectioning finally. The method disclosed by the invention can be used for solving the problem that, the sample pretreatment process is complicated, the toxicity of used medicaments is strong, and the ultrastructure in a PAOs cell is difficult to clearly display in existing methods; and once the preparation method is adopted for preparing PAOs samples, in TEM observation, the cell structures of bacteria, especially poly-Ps in bacteria, can be clearly and completely revealed.

Description

technical field

[0001] The invention belongs to the field of preparation of TEM ultra-thin section samples, in particular to a method for preparing phosphorus-accumulating bacteria transmission electron microscope ultra-thin section samples. Background technique

[0002] Phosphorus accumulating organisms (PAOs) play a key role in the process of biological phosphorus removal. The phosphorus removal function of PAOs is closely related to the number of intracellular polymers such as polyphosphate particles (poly-Ps). In view of the importance of intracellular poly-Ps in PAOs, qualitative and quantitative analysis and determination are often carried out, usually by staining and observing the distribution of poly-Ps in PAOs with the help of optical or electron microscopes (staining conditions for intracellular polymers of PAOs Optimization and comparison of dyeing methods [J]. Environmental Science and Technology, 2014, (2): 1-6). However, the resolution of the optical microscop...

Images