Texture identification in liquid crystal-protein droplets using evaporative drying, generalized additive modeling, and K-means Clustering.

Sessile drying droplets manifest distinct morphological patterns, encompassing diverse systems, viz., DNA, proteins, blood, and protein-liquid crystal (LC) complexes. This study employs an integrated methodology that combines drying droplet, image texture analysis (features from First Order Statistics, Gray Level Co-occurrence Matrix, Gray Level Run Length Matrix, Gray Level Size Zone Matrix, and Gray Level Dependence Matrix), and statistical data analysis (Generalized Additive Modeling and K-means clustering). It provides a comprehensive qualitative and quantitative exploration by examining LC-protein droplets at varying initial phosphate buffered concentrations (0x, 0.25x, 0.5x, 0.75x, and 1x) during the drying process under optical microscopy with crossed polarizing configuration. Notably, it unveils distinct LC-protein textures across three drying stages: initial, middle, and final. The Generalized Additive Modeling (GAM) reveals that all the features significantly contribute to differentiating LC-protein droplets. Integrating the K-means clustering method with GAM analysis elucidates how textures evolve through the three drying stages compared to the entire drying process. Notably, the final drying stage stands out with well-defined, non-overlapping clusters, supporting the visual observations of unique LC textures. Furthermore, this paper contributes valuable insights, showcasing the efficacy of drying droplets as a rapid and straightforward tool for characterizing and classifying dynamic LC textures.

Drying of bio-colloidal sessile droplets: Advances, applications, and perspectives.

Drying of biologically-relevant sessile droplets, including passive systems such as DNA, proteins, plasma, and blood, as well as active microbial systems comprising bacterial and algal dispersions, has garnered considerable attention over the last decades. Distinct morphological patterns emerge when bio-colloids undergo evaporative drying, with significant potential in a wide range of biomedical applications, spanning bio-sensing, medical diagnostics, drug delivery, and antimicrobial resistance. Consequently, the prospects of novel and thrifty bio-medical toolkits based on drying bio-colloids have driven tremendous progress in the science of morphological patterns and advanced quantitative image-based analysis. This review presents a comprehensive overview of bio-colloidal droplets drying on solid substrates, focusing on the experimental progress during the last ten years. We provide a summary of the physical and material properties of relevant bio-colloids and link their native composition (constituent particles, solvent, and concentrations) to the patterns emerging due to drying. We specifically examined the drying patterns generated by passive bio-colloids (e.g., DNA, globular, fibrous, composite proteins, plasma, serum, blood, urine, tears, and saliva). This article highlights how the emerging morphological patterns are influenced by the nature of the biological entities and the solvent, micro- and global environmental conditions (temperature and relative humidity), and substrate attributes like wettability. Crucially, correlations between emergent patterns and the initial droplet compositions enable the detection of potential clinical abnormalities when compared with the patterns of drying droplets of healthy control samples, offering a blueprint for the diagnosis of the type and stage of a specific disease (or disorder). Recent experimental investigations of pattern formation in the bio-mimetic and salivary drying droplets in the context of COVID-19 are also presented. We further summarized the role of biologically active agents in the drying process, including bacteria, algae, spermatozoa, and nematodes, and discussed the coupling between self-propulsion and hydrodynamics during the drying process. We wrap up the review by highlighting the role of cross-scale in situ experimental techniques for quantifying sub-micron to micro-scale features and the critical role of cross-disciplinary approaches (e.g., experimental and image processing techniques with machine learning algorithms) to quantify and predict the drying-induced features. We conclude the review with a perspective on the next generation of research and applications based on drying droplets, ultimately enabling innovative solutions and quantitative tools to investigate this exciting interface of physics, biology, data sciences, and machine learning.

Temperature and Concentration Dependence of Human Whole Blood and Protein Drying Droplets.

The drying of bio-colloidal droplets can be used in many medical and forensic applications. The whole human blood is the most complex bio-colloid system, whereas bovine serum albumin (BSA) is the simplest. This paper focuses on the drying characteristics and the final morphology of these two bio-colloids. The experiments were conducted by varying their initial concentrations, and the solutions were dried under various controlled substrate temperatures using optical and scanning electron microscopy. The droplet parameters (the contact angle, the fluid front, and the first-order image statistics) reveal the drying process's unique features. Interestingly, both BSA and blood drying droplets' contact angle measurements show evidence of a concentration-driven transition as the behavior changes from non-monotonic to monotonic decrease. This result indicates that this transition behavior is not limited to multi-component bio-colloid (blood) only, but may be a phenomenon of a bio-colloidal solution containing a large number of interacting components. The high dilution of blood behaves like the BSA solution. The ring-like deposition, the crack morphology, and the microstructures suggest that the components have enough time to segregate and deposit onto the substrate under ambient conditions. However, there is insufficient time for evaporative-driven segregation to occur at elevated temperatures, as expected.

Concentration-driven phase transition and self-assembly in drying droplets of diluting whole blood.

Multi-colloidal systems exhibit a variety of structural and functional complexity owing to their ability to interact amongst different components into self-assembled structures. This paper presents experimental confirmations that reveal an interesting sharp phase transition during the drying state and in the dried film as a function of diluting concentrations ranging from 100% (undiluted whole blood) to 12.5% (diluted concentrations). An additional complementary contact angle measurement exhibits a monotonic decrease with a peak as a function of drying. This peak is related to a change in visco-elasticity that decreases with dilution, and disappears at the dilution concentration for the observed phase transition equivalent to 62% (v/v). This unique behavior is clearly commensurate with the optical image statistics and morphological analysis; and it is driven by the decrease in the interactions between various components within this bio-colloid. The implications of these phenomenal systems may address many open-ended questions of complex hierarchical structures.

A comparative study of the drying evolution and dried morphology of two globular proteins in de-ionized water solutions.

Pattern formation in drying protein droplets continues to attract considerable research attention because it can be linked to specific protein-protein interactions. An extensive study of the drying evolution and the final crack patterns is presented, highlighting the concentration dependence (from 1 to 13 wt%) of two globular proteins, lysozyme (Lys) and bovine serum albumin (BSA), in de-ionized water. The drying evolution starts with a constant contact radius mode and shifts to a mixed mode where both fluid front and contact angle changes. The contact angle monotonically decreases, whereas, the fluid front exhibits two regimes: an initial linear regime and a later non-linear regime. Unlike the linear regime, the non-linear regime is faster for Lys droplets. This results in the formation of a "mound"-like structure in the central region. A new feature, a "dimple" is observed in this mound which is found to be dependent on the initial concentration. The different crack morphology of BSA and Lys depends strongly on the initial state of the solution and can be interpreted using a simple mechanical model. In fact, when dried under uniform conditions (surface, humidity, temperature, droplet diameter, etc.), the evolution and the final pattern displays as a fingerprint of the initial state.