Transcript
DR KATJA RIETDORF:
This briefing video provides an overview of the use of the digital fluorescence microscope. Full instructions for the ‘Fluorescence Microscopy Activity – Your journey into the cell’ are found in your Practical Workbook.
In the previous topic, you learned how to use a digital microscope. Here you will encounter a digital microscope with the added functionality of using fluorescent light to inspect your cells. The workbook contains an introduction to fluorescence microscopy and explains how this works. You should read these sections if you are unfamiliar with using a fluorescence microscope. It will help you to understand how the differently coloured images that you will see in this activity have been captured using light of different wavelengths in separate channels to excite the probes used for staining the cells.
On opening the microscope, you’ll see an image fairly similar to that of the digital microscope you have already used. In the slide box on the right hand side, you find a description of each slide. You see a numbered list of the available channels. This contains the details on which fluorescent probes or antibodies have been used to obtain the respective image. It also lists the information which channels have been combined into merged views. To display the different channels, click on the numbers at the bottom of the slide description.
In the tutorial slide, channel one shows you the nuclei, channel two the mitochondrial network, and channel three the cytosol and nuclei. The following four channels give you a merged view. Channel four combines nuclei and the cytosol, illustrating that the bright elliptical areas you saw in channel three are in the same place as the nuclei. Channel five is a merged view of nuclei and mitochondria, channel six of the cytosol and mitochondria. This view nicely illustrates that mitochondria are absent from the nucleus.
Finally, channel seven is a merged view of all three channels. Like before, you’ve got points of interest indicated, which take you to specified coordinates in the slides. And you can change between different objectives. As noted in the workbook, additional magnification was used when capturing these images, explaining why you see cellular structures in more detail here than when using the same objectives in the earlier digital microscope activity.
The slide box contains a selection of slides, showing examples of organelles and proteins that you will encounter when studying this topic. As you have seen before, there’s a tool box on the left hand side and a camera button to capture images for your records and assessment activities.