Quantitative Analysis of Mica-Adsorbed HRP

The heights of mica-adsorbed HRP were determined as the heights corresponding to the maxima of the respective distributions of the mica-adsorbed objects with heights ρ(h) as reported in [41 (link)]:
where N_h is the number of AFM-visualized mica-adsorbed objects of the height h, while N is the total number of mica-adsorbed objects visualized by AFM.
The experimentally obtained dependence (1) was then approximated using the Gaussian function: $$\rho \left(h\right)={{\displaystyle \sum }}^{}{\rho }_i\left(h\right)={\displaystyle \sum }_{i=1}^2\frac{A{e}^{\frac{-4 ln(2){(h-h_c)}^2}{w^2}}}{w\sqrt{\frac{\pi }{4 ln(2)}}}$$
where A, h_c, and w are the parameters varied upon approximation. The maxima of the ρ(h) distributions were calculated as the maxima of the approximation Function (2) for each distribution. The analysis of the approximation of the experimentally obtained ρ(h) distributions is based on a χ² criterion.
The initial processing of the AFM images obtained (second-order plane subtraction) and the data export to ASCII format were performed with an Image Analysis software (NT-MDT, Zelenograd, Russia) supplied with the atomic force microscopes. The number of objects, visualized in the AFM images obtained, was calculated with a specialized Recognite software (developed in Institute of Biomedical Chemistry in collaboration of Moscow Engineering Physical Institute MEPhI).