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O-MUG

Comprehensive Course on Fluorescence Microscopy


Advanced Widefield Techniques

Approximate Time: 1-2hrs
Pre-requisite: Widefield Microscope Training
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This is an extension of the widefield microscope training course. It provides training on a specific technique, such as tiling, darkfield microscopy, and timelapse imaging. These techniques are not available on all the widefield microscopes. Please come talk to us about your application before booking a training, so that we can help you choose the right instrument. Here are some of the techniques available:

  • Tiling (Olympus Upright): Specialized software is used to automatically acquire adjacent images across a user-defined tissue area. The microscope is also fitted with motorized filters, so that multiple fluorphores can be acquired at each position. The images are then stitched together to create a composite image of the area of interest. The training will include:
    • Software setup (calibrations, setting parameters, etc.)
    • How to choose the tissue region to tile
    • Stitching of the final image
  • Timelapse imaging (AxioObserver): Is used to acquire a sequence of images in time to observe a dynamic event. The training will include:
    • Stage-top incubator setup (if required)
    • How to set up a time sequence in the software (choosing number of images, delay, channels, etc.)
    • Saving the sequence as a movie file (including setting playback speeds/frame rates, etc.)
  • Darkfield (Olympus Upright): is a transmitted light technique that can be used to achieve greater contrast in images of unstained, transparent specimens. It is often used to reveal external morphology: outlines, boundaries, edges and refractive index gradients. Some examples of specimens that are ideal for darkfield microscopy include zooplankton, diatoms, small insects, fibers, cultured cells as well as non-biological specimens having porosity differences or refractive index gradients. The training will include:
    • Theory of darkfied illumination
    • Setting up/optimizing the microscope components for darkfield illumination
  • Polarization (Olympus Upright): is a transmitted light technique that can be used to achieve greater contrast in images of unstained, transparent specimens. It is often used to reveal external morphology: outlines, boundaries, edges and refractive index gradients. Some examples of specimens that are ideal for darkfield microscopy include zooplankton, diatoms, small insects, fibers, cultured cells as well as non-biological specimens having porosity differences or refractive index gradients. The training will include:
    • Theory of polarization microscopy
    • Setting up/optimizing the microscope components for polarization microscopy

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