Super-precise force clamp experimental method based on biofilm mechanical probe system

Abstract

The invention discloses a super-precise force clamp experimental method based on a biofilm mechanical probe system. The force clamp experiment involves a probe end and a target end. The probe end comprises a probe small ball and red blood cells. The super-precise force clamp experimental method is characterized in that: in a biofilm mechanical probe force clamp experiment, a reference small ball is added at the probe end, the reference ball is adhered to the end part of a first micropipette for sucking red blood cells through strong interaction between protein molecules, the relative distancebetween the boundary of the reference ball close to one side of the red blood cells and the boundary of the probe ball close to one side of the red blood cells is tracked in real time, and the movement of the target end is controlled through adopting a feedback algorithm to conduct a force clamp experiment. By adding the reference sphere and developing a ''double-boundary'' force tracking mode based on a biofilm mechanical probe system, the precision of force acting on molecular bonds is remarkably improved, precise and stable acting force can be applied to the molecular bonds with a low dissociation rate, the recording time limit of the molecular bond binding time is remarkably prolonged, and long-time recording is carried out.

Description

technical field [0001] The invention relates to a quantitative detection method for detecting the dissociation rate of interaction between biological macromolecules, in particular to a super quasi-force clamp experimental method based on a biofilm mechanics probe system. Background technique [0002] With the development of single-molecule force spectroscopy, technologies such as atomic force microscopy, optical tweezers, magnetic tweezers, and biofilm mechanical probes have analyzed the regulation of various biological forces on single-molecule bonds at the single-molecule level. The biofilm mechanical probe (BFP) system has the advantages of small elastic coefficient, low operation difficulty, and easy operation on living cells, and is especially suitable for in situ detection of biomacromolecular interactions on the surface of living cells. As an important part of the dynamic force spectrum experiment, the "force clamp" experiment quickly loads the force acting on the mol...

AI Technical Summary

Problems solved by technology

[0003] Although the biomechanical force clamp experiment has detected a variety of transient molecular interactions with fast dissociation rates, it still cannot be applied to the study of intermolecular interactions with slow dissociation rates

This is because the force feedback control function is lacking in the detection process, and various disturbances will cause the force value acting on the molecular bond to shift continuously during the long bonding time of the molecular bond; in addition, during the ultra-long bonding time of the molecular bond During the recording process, the probe end will also drift as a whole. Even if the position of the ball at the probe end is maintained at the set position, the actual force acting on the molecular bond may have a large deviation, which will eventually result in poor measurement accuracy. , and even draw wrong conclusions

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