Digital Optical and Scanning Probe Microscopy for Inspection and Manipulation of Biocells

Chapter
Part of the Bioanalysis book series (BIOANALYSIS, volume 2)

Abstract

As biomedical interest has progressed to the study of biological tissues and single cells investigations of the different biomaterials involved require very careful use of fine-scale measurement methods, Atomic Force Microscopy (AFM) and Dynamic Laser Speckle (DLS) being reported here. AFM and DLS are complex experimental systems with the functions of scanning probe and optical microscopy. A special optical system makes it possible to visualize the objects and position the probe within microscale dimensions. AFM is used both for visualization and identification of the local adhesion and viscoelastic properties of biological cells, and for manipulation of the cell by means of varying the load being applied to it. Additional information about cellular activity could be obtained by laser probing via DLS of the living tissues being studied. These techniques, AFM and DLS, greatly enhance the potential for measurements and open a new field of experiments in cell biology. The purpose of this chapter is to show the application of AFM and DLS to studies of biological cells, namely measurement of the motility of general cells in living tissues and the elastic modulus of a single cell membrane, as well as identifying the forces causing membrane damage. The time-space cross-correlation analysis of the temporal evaluation of the dynamic biospeckle patterns is shown to be a means of real time flow visualization of the microcirculation of blood in living tissue. Digital processing of biospeckle pattern records yields 2D maps exhibiting the temporal and spatial variations in subskin blood flow. This could be used for biomedical diagnostic purposes, e.g., for detecting microscale deviations from the normal case. Three methods of evaluating dynamic speckle patterns are described. Both decorrelation and autocorrelation analyses have been realized in real time mode, when a total digital specklegram treatment was performed during the time interval between successive frames (40 ms). Results in the form of 2D maps of subskin blood flux were visualized on the PC monitor with a frequency of 25 Hz.

Keywords

Migration Adenocarcinoma Heparin Boiling Autocorrelation 

Notes

Acknowledgments

The authors wish to thank Dr. Serguei Rubnikovich (Belarus Medical State University, Minsk), Dr. Nikolai Bazylev, and Ms. Olga Meleeva (Heat and Mass Transfer Institute) for the help in images treatment and manuscript preparation as well as the National Academy of Sciences and Foundation for Basic Research of Belarus for partial financial support of the work with grants and projects “Energy Efficiency EE 1.6.1,” T11MC-023, T10-029.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.A.V. Likov Heat and Mass Transfer, Institute of National Academy of Sciences of BelarusMinskBelarus

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