Cell culture systems provide several advantages as models of more complex biological systems, including their potential for high-throughput screening, reproducibility, cost-effectiveness, reduced ethical considerations, well-documented protocols, and their ability to be easily manipulated. There are over 4000 well-characterised, readily available cell lines serving as models for human disease and development, which can be studied by researchers using a variety of imaging applications. Super Resolution Microscopy (SRM) Going beyond the Limits of Light Light-sheet fluorescence microscopy enables imaging of larger, multicellular samples, such as organoids and whole organisms (e.g., insects, plants, and animals ) that may not be adequately captured in a timely manner using single or multi-photon microscopy, although this does require tissue-clearing methodology to create optically transparent samples.
The thickness of the sheet of light determines the thickness of the optical section captured, and the sample can be rotated and moved through this plane to produce a three-dimensional tomographic image. Emitted light is then detected perpendicular to the excitation sheet, unlike a confocal microscope, which detects the emitted signal in the opposite direction to the excitation beam. Light-sheet microscope operation differs from confocal microscopy, as samples are excited using a sheet of light from the side, as opposed to a focussed beam of light from the top or bottom of a sample. Fluorescence Microscope Hardware SystemsĪnother recent technique, used to image larger tissue samples quickly and efficiently, is light-sheet fluorescence microscopy (LSFM). A helpful flowchart ( Figure 1) is also provided to facilitate researchers to select the most appropriate combination of hardware, biological system, and imaging agent to achieve the best possible outcome in their imaging experiments. The advantages and limitations for each factor will be discussed to allow the reader to make informed decisions when contemplating experimental design. This review aims to summarise the three main considerations when fluorescence microscopy experiments are employed: (1) current hardware availability (2) the different biological models applicable for fluorescence microscopy and (3) the plethora of fluorescent probes available. A misunderstanding of some of these disciplines by researchers from other areas often leads to poor experimental design and research outcomes. The rapid advances in the field of fluorescence microscopy have required a collaborative effort from physicists and engineers developing microscopy hardware, to chemists that continually develop and refine novel fluorescent probes, and biologists who utilise these tools to investigate the biological functions of diverse specimens. The discovery of novel fluorescent markers has driven the development of fluorescence-competent microscope technologies, further enabling researchers to discover and understand the intricate dynamics of subcellular biology and their detailed mechanics at ever-higher spatial and temporal resolutions ( Figure S1). The use of fluorescently-labelled antibodies in the 1940s enabled precision visualisation of target structures in cells, and with the Nobel prize winning discovery of green fluorescent proteins (GFP) in the 1960s and their subsequent development as a genetic tag, the field has evolved to permit targeted live cell imaging. Initially, auto-fluorescent specimens were visualised with fledgling fluorescence microscopes, until the introduction of fluorescent stains in the 1930s, which enabled non-fluorescent specimens to be visualised. If you get lost, written instructions are provided at the center of each table as well as on the wall.Since the inception of fluorescence microscopes in the early 1900s, their use as a research tool for observing discrete subcellular structures and processes has grown immensely. We will also help you with the selection of stain and paint color options. The instructor will guide you through the process, from start to finish.
Larger pieces and furniture projects may last up to 3.5 hours. A typical class will last for 2-2.5 hours. Once everyone has settled in, we will begin class. If you also wish to place your purse/handbag away from the work table, there is a designated area where you can safely place your things. In order to protect your clothing, you will want to put on the apron provided. If you requested to be seated next to a friend, you should find their design in the spot next to you. Never been to a wood and paint studio? No worries, you are not alone! Here is what to expect when you arrive:īoth your wood and your personalized stencil will be placed on a work table at a reserved spot.
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