Engaging new audiences with imaging and microscopy

ABSTRACT In this Spotlight, we hear first-hand accounts from five scientists and educators who use microscopy and imaging to engage, entertain, educate and inspire new audiences with science and the field of developmental biology in particular. The ‘voices’ that follow each convey each authors' own personal take on why microscopy is such a powerful tool for capturing the minds, and the hearts, of scientists, students and the public alike. They discuss how microscopy and imaging can reveal new worlds, and improve our communication and understanding of developmental biology, as well as break down barriers and promote diversity for future generations of scientific researchers.

FACT For Fast And Three-Dimensional Imaging of Intact Tissues

Free of Acrylamide Sodium Fast Free-of-Acrylamide Clearing Tissue (FACT) is a developed technique using no acrylamide for clearing tissues. As the lipid removal normally is a harmful process and it causes loss of biological molecules such as proteins and on the other hand is crucial for transparency and efficient antibody staining throughout the whole tissue especially for microscopy and imaging, the FACT technique is suitable since it makes chemical bonding of membrane and intracellular proteins with the extracellular matrix creating a massive three-dimensional (3D) matrix and structural support to fortify the tissue during processing. Compared to other acrylamide-based techniques, FACT requires less labor, toxic, and harmful chemicals. Here we describe protocols encompassing every angle and dimension of the FACT protocol for antibody staining and imaging of whole-cleared tissues while preserving the structure and increasing the image quality. The entire protocol includes tissue perfusion, fixation, clearing, antibody staining, Refractive Index Matching (RIM), microscopy, and imaging; this timing varies due to the size, weight, different kind of tissues and the type of immunostaining. This technique has been favorably performed on different types of tissues for molecular interrogation analysis of large tissues.

Imaging techniques in nanomedical research

About twenty years ago, nanotechnology began to be applied to biomedical issues giving rise to the research field called nanomedicine. Thus, the study of the interactions between nanomaterials and the biological environment became of primary importance in order to design safe and effective nanoconstructs suitable for diagnostic and/or therapeutic purposes. Consequently, imaging techniques have increasingly been used in the production, characterisation and preclinical/clinical application of nanomedical tools. This work aims at making an overview of the microscopy and imaging techniques in vivo and in vitro in their application to nanomedical investigation, and to stress their contribution to this developing research field.

Advances in Surface Plasmon Resonance Imaging and Microscopy and Their Biological Applications

Surface plasmon resonance microscopy and imaging are optical methods that enable observation and quantification of interactions of nano- and microscale objects near a metal surface in a temporally and spatially resolved manner. This review describes the principles of surface plasmon resonance microscopy and imaging and discusses recent advances in these methods, in particular, in optical platforms and functional coatings. In addition, the biological applications of these methods are reviewed. These include the detection of a broad variety of analytes (nucleic acids, proteins, bacteria), the investigation of biological systems (bacteria and cells), and biomolecular interactions (drug–receptor, protein–protein, protein–DNA, protein–cell).

The Recent Advances in Raman Microscopy and Imaging Techniques for Biosensors

Raman microspectroscopy is now well established as one of the most powerful analytical techniques for a diverse range of applications in physical (material) and biological sciences. Consequently, the technique provides exceptional analytical opportunities to the science and technology of biosensing due to its capability to analyze both parts of a biosensor system—biologically sensitive components, and a variety of materials and systems used in physicochemical transducers. Recent technological developments in Raman spectral imaging have brought additional possibilities in two- and three-dimensional (2D and 3D) characterization of the biosensor’s constituents and their changes on a submicrometer scale in a label-free, real-time nondestructive method of detection. In this report, the essential components and features of a modern confocal Raman microscope are reviewed using the instance of Thermo Scientific DXRxi Raman imaging microscope, and examples of the potential applications of Raman microscopy and imaging for constituents of biosensors are presented.