Automated fractal microscope for virus-cell monitoring

Abstract number: p654

Fedchuk  O.P., Fedchuk  A.O., Fedchuk  A.S.

Objectives: 

Virus-cell interaction could be monitored in a standard way through direct infected cells counting in the optical luminescent microscope. This way is time- and effort- consuming and requires additional colouring of the cell culture and the sample preparation process takes from 24 to 48 hours. The reliability of this method is not sufficient because it monitor only a small part of the specimen surface. Taking into account the results of our previous experimental investigations in the field, we may suppose that practically every stage of virus-cell interaction could be described in objective quantitative manner using the fractal approach and corresponding original instrumentation.

Methods: 

We have studied the interaction of Herpes simplex virus strain US-1 (HSV-1) with the specific cell culture Hep-2 modified by addition of proteolysis inhibitor E-aminocaproic acid (E-ACA) and Acyclovir taken in various concentrations. The monolayer of the cell culture was placed between the glasses support. The described samples were washed out with Hanks solution and afterwards fixed with ethanol. The fractal microscope included the single-mode and intensity stabilized Spectra-Physics He-Ne laser with 5.0 mW output and wavelength of 0.6328 + 0.01 microns. The diffraction patterns (DP) of the samples were taken with the use of Olympus Z-8080 digital camera and introduced with the use of USB cable into the port of Pentium IV computer. The fractal dimension D of the DP was evaluated automatically in an express manner (2 minutes per sample).

Results: 

The main result of our experiments was, of course, the multi-fractal type of the DP clusters. The registered minimal sizes of the fractal cluster elements were practically about the cell size ~1 mkm and HSV-1 virion ~0.1 mkm. The DP picture on the target is Fourier transform of the object and the real picture of the virus-cell system is obtained as the reverse Fourier transform of the DP.

Conclusions: 

The proposed device and method are applicable and highly competitive in laboratory and clinical antiviral research as well as in the drug design and testing process due to its high sensitivity, express character of data taking and precise quantitative way of data processing.