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Research on B Lymphocyte Morphology and Biomechanics Based on Atomic Force Microscope

Author: WangQiuLan
Tutor: CaiJiYe
School: Jinan University
Course: Analytical Chemistry
Keywords: Atomic force microscopy B lymphocytes Chronic lymphocytic leukemia Ultrastructure Biomechanics
CLC: R318.01
Type: Master's thesis
Year: 2011
Downloads: 51
Quote: 0
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Abstract


Based on atomic force microscopy, combined with quantum dots marking technology, laser scanning confocal microscopy and flow cytometry and other biological detection technologies and density gradient centrifugation, positive and negative magnetic bead separation methods such as sorting method to normal healthy people and chronic lymphocytic leukemia patients for the study of peripheral blood B lymphocytes, in the nanometer scale resolution perspective from two different biophysical properties of cells, i.e. cells nanostructure morphology and membrane, the membrane surface adhesion properties, the Young's modulus , the antigen - antibody specific interaction between the surface of the film such as the distribution of antigen. Mainly obtained the following innovative work: 1) Compared with the normal B-lymphocytes, B-lymphocytes tall chronic lymphocytic leukemia, nano-structure is more rough surface of the membrane, non-specific adhesion and hardness are significantly reduced. CD20 antibodies by modifying the AFM tip measured CD20 antigen - antibody interaction tip than the unmodified antibody measured non-specific adhesion of five times, and based on the force measuring antigen-specific antibody, CD20 molecules derived normal CD20 B lymphocytes membrane is more evenly distributed, there is a small part of the aggregation phenomenon, on the contrary, in CLL CD20 B lymphocytes membrane surface sparse. AFM indicates that the application is available in morphology and mechanical properties significantly distinguish between normal and chronic lymphocytic leukemia B cells for clinical diagnosis of chronic lymphocytic leukemia provide new means, B lymphocytes CD20 antigen CD20 drug distribution can be selected as a reference. 2) With the SAC superantigen stimulation time increases, 1.2h to 72h CLL B lymphocytes volume, diameter and nuclear cytoplasmic ratio increased significantly, 24h appear pseudopodia and immunological synapse phenomenon. During the activation cell surface molecules after nanostructures from loose agglomeration and the final formation of further micro-domains of the change process, the cell size distribution and roughness parameters changed significantly increased cell adhesion. This in-depth understanding of CLL B lymphocyte functional defects in the form of lower activation process such as changes in biomechanical properties provide a visual basis. 3) compared with resting B lymphocytes, antigen stimulation PWM or SAC after 24h molecules B lymphocyte surface markers such as CD69 expression significantly increased in the early activation phase cells, and appears to enhance BCR signaling, cytoskeleton movement and a key protein in the expression of F-actin increases, leading to a series of biological changes in physical properties, including cell morphology height, diameter and volume changes of different degree, Ultrastructure surface adhesion protein aggregation occurs and power is significantly increased by 2 to 3 times and membrane depolarization phenomenon. It is able to in-depth understanding of B lymphocyte activation process provides new information and structure function analysis. These innovative results of biological, clinical, and other aspects of B lymphocyte immune study provides some methods and data reference value.

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CLC: > Medicine, health > Basic Medical > Medical science in general > Biomedical Engineering > General issues > Biomechanics
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