New tools to study biophysical properties of single molecules and single cells

Anais da Academia Brasileira de Ciências

Márcio S. Rocha¹  Oscar N. Mesquita¹ 

[1] ,Universidade Federal de Minas Gerais ICEx Departamento de FísicaBelo Horizonte MG ,Brasil

关键词: optical tweezers;    defocusing microscopy;    single molecules;    macrophages;    water transport;    cell motility;    pinça óptica;    microscopia de desfocalização;    moléculas únicas;    macrófagos;    transporte de água;    motilidade celular;   

DOI  :  10.1590/S0001-37652007000100003

来源: SciELO

【 摘 要 】

We present a review on two new tools to study biophysical properties of single molecules and single cells. A laser incident through a high numerical aperture microscope objective can trap small dielectric particles near the focus. This arrangement is named optical tweezers. This technique has the advantage to permit manipulation of a single individual object. We use optical tweezers to measure the entropic elasticity of a single DNA molecule and its interaction with the drug Psoralen. Optical tweezers are also used to hold a kidney cell MDCK away from the substrate to allow precise volume measurements of this single cell during an osmotic shock. This procedure allows us to obtain information about membrane water permeability and regulatory volume increase. Defocusing microscopy is a recent technique invented in our laboratory, which allows the observation of transparent objects, by simply defocusing the microscope in a controlled way. Our physical model of a defocused microscope shows that the image contrast observed in this case is proportional to the defocus distance and to the curvature of the transparent object. Defocusing microscopy is very useful to study motility and mechanical properties of cells. We show here the application of defocusing microscopy to measurements of macrophage surface fluctuations and their influence on phagocytosis.

【 授权许可】

CC BY   
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