A three-dimensional surface reconstruction method suitable for biofilm
A technology of three-dimensional curved surface and curved surface reconstruction, applied in the field of biology, can solve the problems of limited thickness, unable to penetrate dense biofilm tissue, unable to obtain ideal characterization results, etc., and achieve the effect of high reduction degree
Active Publication Date: 2022-05-27
DALIAN UNIV
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Problems solved by technology
However, for biofilms with complex three-dimensional structures and thick biofilms, the characterization of their physical forms has always been one of the problems that plague the majority of researchers.
Scholars at home and abroad use confocal laser microscopy (CLSM) to measure the three-dimensional structure of biofilms. The limited amount of light transmitted cannot penetrate the dense biofilm tissue, so ideal characterization results cannot be obtained
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Embodiment 1
[0075] (1) Making biofilm slices
[0076] (1) Cleaning and sampling
[0077] The biofilm carrier was washed with PBS to remove the nutrient solution contained on its surface and inside, and the biofilm was scraped into the petri dish with tools;
[0078] (2) Fixed
[0079] The biofilm samples were fixed in 2.5v / v% glutaraldehyde solution and placed at 4°C for 24h;
[0080] (3) Rinse
[0081] Take out the sample, absorb the excess fixative on the surface with absorbent paper, rinse three times with PBS phosphate buffer solution of 0.2mol / L, pH 7.2;
[0082] (4) Dehydration
[0083] The samples were dehydrated with 50%, 75%, 85%, 90% and 95% ethanol solutions respectively for 15min; then treated with 100% ethanol twice, 20min each time; (5) Permeabilization
[0084] Use 0.5v / v% TritonX-100 / PBS solution for 10 min at 4°C to increase the permeability of cell membrane, and wash with 0.2mol / L PBS phosphate buffer solution 3 times;
[0085] (6) Wax dipping and embedding
[008...
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Abstract
The invention relates to a three-dimensional curved surface reconstruction method suitable for biological membranes, belonging to the technical field of biology. The main technical solutions are as follows: cleaning and sampling, fixation, rinsing, dehydration, permeabilization, wax dipping and embedding, slicing, sticking and baking, dewaxing, dyeing, storage; adjusting the emitted light wavelength, adjusting the magnification, taking pictures and Image storage; building 3D models. The present invention is aimed at biofilms with a thickness greater than 0.1mm, in order to avoid characterization difficulties caused by dyes and laser beams not being able to effectively enter the interior of biofilms due to too thick biofilms. Using this technology can not only effectively reconstruct the complex physical form of biofilm, but also ensure a high degree of reduction, which provides a practical means for the study of biofilm morphology changes and influencing factors.
Description
technical field [0001] The invention relates to the technical field of biology, in particular to a three-dimensional curved surface reconstruction method suitable for biological membranes. Background technique [0002] Biofilms are one of the main forms of microbial aggregation. Biofilms are layered aggregates formed by microorganisms and extracellular polymers attached to solid surfaces, with a certain thickness and complex three-dimensional morphology. Biofilms are ubiquitous in nature, but human research on biofilms begins with biofilms attached to reactors and carriers in biofilm wastewater treatment processes. The biofilm system is usually composed of four parts: the bulk liquid, the boundary layer, the biofilm and the carrier. figure 1 shown. The formation of biofilm includes two processes of attachment and immobilization of microorganisms on the carrier surface, which are affected by factors such as fluid flow, substrate type and temperature. Biofilms not only hav...
Claims
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IPC IPC(8): G01N21/64G01N1/30G01N1/36G01N1/28G01N1/34G06T17/20
Inventor韩越梅
OwnerDALIAN UNIV