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In recent decades, mass spectrometry-based lipidomics has provided a fertile environment for scientific investigations of biochemical and mechanistic processes in biological systems. Notably, this approach has been used to characterize physiological and pathological processes relevant to the central nervous system by identifying changes in the sphingolipid content in the brain, cerebral spinal fluid, and blood plasma. However, despite a preponderance of studies identifying correlations between specific lipids and disease progression, this powerful resource has not yet substantively translated into clinically useful diagnostic assays. Part of this gap may be explained by insufficient depth of the lipidomic profiles in many studies, by lab-to-lab inconsistencies in methodology, and a lack of absolute quantification. These issues limit the identification of specific molecular species and the harmonization of results across independent studies. In this chapter, we contextualize these issues with recent reports identifying correlations between brain lipids and neurological diseases, and we describe the workflow our group has optimized for analysis of the blood plasma sphingolipidome, adapted to the characterization of the human brain tissue., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

In the process of constructing and characterizing the whole cell biosensor for Vibrio cholerae detection, two main techniques have been employed-DNA assembly using the Gibson isothermal assembly reaction was used for the assembly of the PCRed plasmid fragments (DNA parts), and microplate fluorescence readings were used for bacterial strain characterization. The general workflow can be summed up as: the in silico designed DNA fragments were assembled by isothermal assembly to be later transformed into Escherichia coli that, in turn, was characterized using the microplate reader. As fine-tuning of the sensor design was required, the process was repeated iteratively until the final strain was created with desired characteristics. This chapter describes in detail this workflow for different constructs which finally led to the creation of the first whole cell biosensor in E. coli for V. cholerae detection.

Mesenchymal stem cells (MSCs) are multipotent cells that have the potential to differentiate into various mesenchymal lineages in vitro and in vivo. Due to their availability from tissues such as bone marrow, synovium, fat, and muscle, and their highly proliferative capacity, MSCs have evoked interest as a potential cell source for repair and regeneration of various types of tissues. Characterization by the expression of a panel of surface markers and the ability of MSCs to undergo multilineage differentiation is the benchmark for identifying this stem cell population. In this chapter, the protocols for the differentiation of MSC to chondrogenic, osteogenic, and adipogenic lineages and histological and immunostaining protocols for confirming trilineage differentiation of the MSC cells are described.