Areas of activity in biofilms through the biospeckle and the spectral domain

Abstract

The dynamic laser speckle or biospeckle laser has been used to analyze the activity of biological and non-biological material by means of various statistical techniques and image processing. However, a challenge to adopt this technique is the ability to identify, in the same material, an area of low activity immersed in an environment of a higher activity. This work was carried out to evaluate the spectral approach associated to biospeckle laser technique as an alternative to identify distinct activities areas in the same material. Biofilm samples, which present well known protocols to be prepared, and a simpler structure than vegetal and animal tissues, were prepared with potato starch and corn starch with areas of different levels of moisture and were analyzed using the biospeckle laser associated with the wavelets transform in order to evaluate the data in the spectral domain. The effect of a black or white background below the samples was also tested. The image analysis was conducted using Generalized Difference and Fujii techniques before and after the implementation of the wavelets transform producing the filtration of the data. The results allowed the visualization of different activities areas in different frequency bands. The areas of activity were presented clearer than the traditional procedures without filtering. A new way to present the results of the biospeckle and the frequency domain information was proposed to enhance the visualization of a whole picture. It was also noted that the greatest contrast between areas of different activity were promoted by materials of different compositions. In some experimental configurations there were possible to tag the relationship between the frequency and depth of the active or inactive material. The influence of the color, black or white, of the background was also noticed in the results, but with white background better in some configurations and with the black better in others.