# Application of Organic-Inorganic Hybrids in Chemical Analysis, Bio- and Environmental Monitoring
## 4 The Role of Nanoanalytics and Nanometrology in Biosensors/Nanobiosensors Development
Unfortunately, nanometrology today is exclusively based on measurement of geometrical features, viz., size, shape, and roughness of an inorganic component at the nanoscale [86; 87; 88; 89; 90; 91; 92; 93], Figure 6. However, to control the roughness (the disorder in the layers) or the shape at nano-scale only makes little sense as compared to the significance of these parameters at the macro- or micro-scales. It should be noted that measuring at the nanoscale is a technological and scientific challenge that makes metrology at the nanoscale special. Thus, there is a great difference in metrological requirements toward macro-, micro-, and nano-scaled objects. Therefore, the measurement techniques and metrological strategies developed for bulk materials cannot be simply applied for standardization of nano-based structures. Instead, a strong adaption or optimization of the conventional methods and development of the novel methodological approaches is highly desirable. Nanoanalytics, as a global scientific approach, play a great role in this process [96; 97]. The term nanoanalytics was initially introduced by Prof. [PERSON] [98; 99]. Later on, the main concepts, elements, and peculiarities of the nanoanalytics were re-reviewed and summarized by Prof. [PERSON] et al. [100]. Recently, the role of nanoanalytics and nanoanalysis as a tool in the development of the modern OInH was highlighted [97]. It is important to stress that two different conceptual strategies are distinguished in nanoanalytics; the first one refers to the methods applied for studying of elemental composition of nano-based hybrids. Another one and more seldomly used strategy in nanoanalytics is to combine several functions in one methodology [98; 99]. Thus, the tandem of several techniques and appropriate optimized methodologies, i.e., scanning electron microscopy (SEM) coupled with energy dispersive X-ray microanalysis (EDX), transmission electron microscopy (TEM), atomic force microscopy (AFM), etc., can be readily applied at once for analysis of OInH to get closer insights on the nature of the redistracted analytical signals from the targeted nano-objects [97]. Remarkably, the great challenge in nanoanalytics belongs to the nanoanalysis of the OInH with polymeric or biological component. Thus, the techniques with nanoscale lateral resolution, such as standard SEM or TEM techniques usually provide no chemical information [99]. At the same time, there is no single analytical method or methodology which is able to do this. Therefore, the analyst  
Figure 6: The relative relationship between different geometric components in conventional (**A**), micromechanics (**B**), and nanoengineering (**C**). Adapted with permission from ref. [94; 95]. working with OIH/OInH has to use and optimize a set/tandem of several techniques of molecular analysis and then summarize their analytical merit. Thus, typically the tandem between Fourier transform infrared spectroscopy (FTIR), RAMAN spectroscopy, surface plasmon resonance (SPR), and mass spectrometry (MS)-based techniques can be applied [97]. However, these methods are unable to visualize/image single biomolecules or a targeted polymer with appropriate accuracy at nano-scale and do not allow a local activation or manipulation of such molecules directly in buffer solutions or in real matrices. In some cases, the above-mentioned problem can be party solved by means of an AFM-based methodology (i.e., local activation or manipulation of biomolecules). This method can be seriously considered as a crucial nanoanalytical tool for characterization of OInH in terms of surface homogeneity (Figure 7) and three-dimensional visualization of polymers or biomolecules (Figure 8). However, in some cases AFM does not allow to distinguish surface homogeneity between the probe and the artifacts. Also, with regards to OInH analysis, AFM provides analytical information on the organic- or bio- component intermixed with the inorganic component over the template surfaces.