Method for detecting mechanical characteristics of red blood cells based on atomic force microscope

Abstract

The invention provides a method for detecting the mechanical characteristics of red blood cells based on an atomic force microscope, which relates to the technical field of biophysics. The technical scheme adopted in the invention is as follows: a round coverslip is used to fix red blood cells; and the method specifically includes the following steps: preparing red blood cell suspension, preprocessing the round coverslip, preparing an atomic force microscope sample, and using the atomic force microscope to detect the mechanical characteristics of the red blood cells in a liquid environment. The red blood cells are fixed by the round coverslip, so that the red blood cells can still be maintained in a physiological state, and the maintenance of the normal morphology and function of the red blood cells is very obvious. Necessary disinfection and innocuous treatment is carried out on the commonly used round coverslip, so that there is no additional drug effect and toxic effect on the red blood cells, and the morphology and function of the cells will not be affected. The red blood cells treated by the round coverslip can be stably attached, the mechanical characteristics of the red blood cells can be independently detected without any other influence or interference, and the real physiological characteristics of the cells can be reflected.

Description

technical field [0001] The invention relates to the technical field of biophysics, in particular to a method for detecting the mechanical characteristics of red blood cells. Background technique [0002] The Atomic Force Microscope (AFM), which appeared in the 1980s, can image the surface morphology and detect the mechanical properties of living cells with extremely high sensitivity and resolution in a liquid environment. The mechanical properties of cells include elasticity, Viscosity and hardness etc. Through the changes of cell elasticity and hardness, the changes of cells in the process of physiology and pathology can be deduced. The study of biological cell mechanics is closely related to many biological processes. The organs, tissues, cells and biomacromolecules of the body undergo corresponding morphological and functional changes under a certain range of mechanical effects. This is the body’s response process to mechanical stimuli, and the body Adaptation to a cert...

AI Technical Summary

Problems solved by technology

2. The internal structure of the sample is not available

AFM can obtain more surface information than other microscopic techniques, but cannot observe the internal structure of cells like Transmission Electron Microscopic (TEM) or near-field optical microscopy

3. Recognition of scanned samples

AFM can only obtain the topography of the sample surface, but cannot identify the substance on the sample surface, so AFM is suitable for scanning a single substance, and it is difficult for AFM to distinguish them directly from the shape of the mixture

4. Scan rate problem

Many biochemical reactions are completed instantaneously and cannot be observed by AFM

5. Since the accuracy of the force applied to the AFM cantilever is affected by external factors such as thermal drift, etc., this drift can only be artificially offset by smoothing, etc.

The most important impact of such treatment on red blood cells is: drying treatment causes red blood cells in a physiological state to lose their normal biological characteristics, properties and functions, and the results obtained under this condition cannot reflect the real RBC shape and function

Secondly, as a chemical substance, no matter how small the dosage is, it will have more or less impact on the biological characteristics or functions of the cells, and the impact on the experimental results is also unavoidable.

Finally, the treatment method of glutaraldehyde fixation can increase the viscosity of the surface of the sample to be tested, causing the AFM probe to be affected by the viscous force and cannot work normally.