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Skin is vulnerable to various external insults such as burn, severe injury, or inflammation, which necessitates a better strategy for wound repair. Mesenchymal stem cells (MSCs) can self-renew and differentiate into various supporting tissues including cartilage, bone, muscle, and adipose tissue. Along with their unique multipotent capacity, they secrete various paracrine mediators such as growth factors, cytokines, and membrane-enclosed particles called extracellular vesicles (EVs). Herein, we discussed the general traits of EVs such as cell-to-cell communicator, and highlighted the recent preclinical outcomes, with a focus on the application of MSC-derived EVs in wound repair. This chapter provides insights into developing novel strategies for skin wound healing in a cell-free manner.Fluorescence imaging of nucleic acids is a key technology needed to understand gene expression and the resulting changes in cellular function. This chapter focuses on sequence-specific fluorescence imaging of nucleic acids in cells using fluorescent nucleic acids. The design and preparation of fluorescent nucleic acids and their application to fluorescence imaging of intracellular nucleic acids are introduced.Semiconductor nanocrystals (SNCs) are a nano-sized inorganic material. Due to the quantum confinement effect, these crystals exhibit unique optical and electrical properties. This chapter focuses on biological applications of SNCs, ranging from in vitro single-molecule tracking to in vivo fluorescence imaging. Indoximod The following fundamental properties and technical procedures related to SNCs are also described structures of SNCs, synthetic procedures and shape control of SNCs, preparation methods of water-soluble SNCs, and conjugation methods of biomolecules.Nanotechnology has been widely applied to medical interventions for prevention, diagnostics, and therapeutics of diseases, and the application of nanotechnology for medical purposes, which is called as a term "nanomedicine" has received tremendous attention. In particular, the design and development of nanoparticle for biosensors have received a great deal of attention, since those are most impactful area of clinical translation showing potential breakthrough in early diagnosis of diseases such as cancers and infections. For example, the nanoparticles that have intrinsic unique features such as magnetic responsive characteristics or photoluminescence can be utilized for noninvasive visualization of inner body. Drug delivery that makes use of drug-containing nanoparticles as a carrier is another field of study, in which the particulate form nanomedicine is given by parenteral administration for further systemic targeting to pathological tissues. In addition, encapsulation into nanoparticles gives the opportunity to secure the sensitive therapeutic payloads that are readily degraded or deactivated until reached to the target in biological environments, or to provide sufficient solubilization (e.g., to deliver compounds which have physicochemical properties that strongly limit their aqueous solubility and therefore systemic bioavailability). The nanomedicine is further intended to enhance the targeting index such as increased specificity and reduced false binding, thus improve the diagnostic and therapeutic performances. In this chapter, principles of nanomaterials for medicine will be thoroughly covered with applications for imaging-based diagnostics and therapeutics.Using the Raman spectroscopic analysis system that gives the chemical information of the biomaterials, classification is performed through the acquisition of fingerprint signals for each cell line, and the basis of the diagnosis is provided. The origin of diagnosis can be clarified by precise analysis through comparison of local signals and morphology in cells, including measurement at tissue level. In this result, normal breast cell line (MCF-10A) and breast cancer cell lines (MDA-MB-231, MDA-MB-453) were characterized using Raman spectroscopy, atomic force microscopy (AFM), and optical microscopy. These three modalities were combined in order to not only separate cancerous and noncancerous cell lines but to analyze their morphological and optical properties. From the results, the inherent optical properties of cancer cells separated from normal cells in terms of local variation were observed. Bright-field (BF) transmission imaging is also compared to the morphological height difference obtained from AFM and is correlated with surface Raman spectra.Spectral reflectometry is a spectroscopic measurement technique based on thin-film interference, which has been widely applied in industries to measure thicknesses of thin dielectric layers at the nanoscale. Recent advances in the understanding of biological nanostructures have opened a new field of spectral reflectometry in biomedicine from molecular level sensing to biomedical imaging. This chapter comprehensively covers the relevant topics on spectral reflectometry in biomedicine from its principle to applications.Optical coherence tomography (OCT) is a three-dimensional (3-D) optical imaging technology that provides noninvasive, micrometer resolution images of structural interiors within biological samples with an approximately 1 ~ 2 mm penetration depth. Over the last decades, advances in OCT have revolutionized biomedical imaging by demonstrating a potential of optical biopsy in preclinical and clinical settings. Recently, functional OCT imaging has shown a promise as angiography to visualize cell-perfused vasculatures in the tissue bed in vivo without requiring any exogenous contrast agents. This new technology termed OCT angiography (OCTA) possesses a unique imaging capability of delineating tissue morphology and blood or lymphatic vessels down to capillaries at real-time acquisition rates. For the past 10 years since 2007, OCTA has been proven to be a useful tool to identify disorder or dysfunction in tissue microcirculation from both experimental animal studies and clinical studies in ophthalmology and dermatology. In this section, we overview about OCTA including a basic principle of OCTA explained with simple optical physics, and its scan protocols and post-processing algorithms for acquisition of angiography. Then, potential and challenge of OCTA for clinical settings are shown with outcomes of human studies.After the emergence of the ultrasound, X-ray CT, PET, and MRI, photoacoustic tomography (PAT) is now in the phase of its exponential growth, with its expected full maturation being another form of mainstream clinical imaging modality. By combining the high contrast benefit of optical imaging and the high-resolution deep imaging capability of ultrasound, PAT can provide unprecedented anatomical image contrasts at clinically relevant depths as well as enable the use of a variety of functional and molecular imaging information, which is not possible with conventional imaging modalities. With these strengths, PAT has achieved numerous breakthroughs in various biomedical applications and also provided new technical platforms that may be able to resolve unmet issues in clinics. In this chapter, we provide an overview of the development of PAT technology for several major biomedical applications and provide an approximate projection of the future of PAT.Live cell imaging provides essential information in the investigation of cell biology and related pathophysiology. Refractive index (RI) can serve as intrinsic optical imaging contrast for 3-D label-free and quantitative live cell imaging, and provide invaluable information to understand various dynamics of cells and tissues for the study of numerous fields. Recently significant advances have been made in imaging methods and analysis approaches utilizing RI, which are now being transferred to biological and medical research fields, providing novel approaches to investigate the pathophysiology of cells. To provide insight into how RI can be used as an imaging contrast for imaging of biological specimens, here we provide the basic principle of RI-based imaging techniques and summarize recent progress on applications, ranging from microbiology, hematology, infectious diseases, hematology, and histopathology.Given the merit of high-resolution cross-sectional imaging, magnetic resonance imaging (MRI) has been utilized in many preclinical and clinical research fields. In addition to T2-weighted imaging for assessing anatomic changes by disease and therapeutic agents, diffusion-weighted imaging, dynamic contrast-enhanced MRI, and MR spectroscopy can provide disease- and drug-specific functional information in both in vivo and ex vivo status. Another advantage of MRI is its ability to bridge the preclinical and clinical experiments as it allows similar study methods and environments between animals and humans. Therefore, MRI can be used as a useful tool for drug development. Investigators have discovered a variety of MRI biomarkers that can quantitatively measure the biological alteration led by disease and treatment. In this chapter, a number of commonly used preclinical MRI biomarkers for drug development will be introduced and discussed, particularly being focused on their value for translational research.Intravital microscopy has emerged as a powerful technique for the fluorescent visualization of cellular- and subcellular-level biological processes in vivo. However, the size of objective lenses used in standard microscopes currently makes it difficult to access internal organs with minimal invasiveness in small animal models, such as mice. Here we describe front- and side-view designs for small-diameter endoscopes based on gradient-index lenses, their construction, their integration into laser scanning confocal microscopy platforms, and their applications for in vivo imaging of fluorescent cells and microvasculature in various organs, including the kidney, bladder, heart, brain, and gastrointestinal tracts, with a focus on the new techniques developed for each imaging application. The combination of novel fluorescence techniques with these powerful imaging methods promises to continue providing novel insights into a variety of diseases.Since their development in the 1960s, immuno-gold techniques have been steadily used in biomedical science, because these techniques are applicable to all kinds of antigens, from viruses to animal tissues. Immuno-gold staining exploits antigen-antibody reactions and is used to investigate locations and interactions of components in the ultrastructure of tissues, cells, and particles. These methods are increasingly used with advanced technologies, such as correlative light and electron microscopy and cryo-techniques. In this protocol, we introduce the principles and technical details of recent advances in this area and discuss their advantages and limitations.Various silica-based fluorescent nanoparticles ((Si-FNP)) with magnetic or metal cores represent a standard class of nanoparticles offering new opportunities for high-resolution cellular imaging and biomedicine applications, such as drug delivery. Their high solubility, homogeneity, biocompatibility, and chemical inertness Si-FNPs make them attractive probes for correlative light and electron microscopy (CLEM) studies, offering novel insights into nanoparticle-cell interactions in detail. In the present chapter, we present a procedure for imaging silica-based fluorescent magnetic core-shell nanoparticles (Si-FMNP) at the single-particle scale in cells. Our method facilitates the acquisition of information on the extracellular and intercellular distribution of nanoparticles and their various interactions with various cellular organelles when cells are cultured and electroporated by NPs. In addition, such information could facilitate the evaluation of the efficacy of nanocarriers designed for drug delivery.