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Testing Light Sheet microscopy

Recently we had a chance to try out the new Zeiss Z1 light sheet microscope at the Institute of Biotechnology (Light Microscopy Unit). Although the planned training was postponed, Swarnalok and I got a quick trial session to examine Nicotiana benthamiana leaved expressing fluorescently labeled host and viral proteins.

In a light sheet microscope the sample is illuminated at with a thin laser sheet. Light emitted from the sample is then detected at a right angle to the direction of the light sheet. The scanning speed is fast compared to a confocal microscope which means the sample is less damaged by the imaging process.

As we had no previous experience with imaging with the light sheet microscope or with protocols of mounting plant samples, we had to improvise. So: when in doubt, use superglue! With the help of Kimmo and Mika of the LMU staff we decided to glue the plant samples onto thin wooden sticks that fit the microscope's sample holder. The superglue-approach worked remarkably well and we even managed to attach a small (~1 cm) young seedling to the stick.

Photo: 1 week-old Nicotiana benthamiana seedling attached with superglue to a stick for imaging with the Zeiss Z1 light sheet system.

We imaged two proteins: a viral protein tagged with red fluorescent protein and an infection-related host protein tagged with yellow fluorescent protein. In addition we imaged chlorophyll autofluorescence. Check out a short video of our scans on Youtube:

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