Confocal Microscopy -  An Introduction

Confocal microscopes are designed to reduce out-of-focus light (aka. blur) from images so only light from the plane of focus can be seen in an image. This allows 3D specimens to be optically sectioned and reconstructed.

Long-term time-lapse makes greater demands of confocal microscope computer hardware and software than simply scanning fixed slides. The following procedures have been recommended for use with the TCS SPE confocal microscopes because they do not have hardware specifically optimised for long-term time-lapse and are more vulnerable to fragmentation, write errors, hangs and other nasty occurrences. They are also advisable when using the SP5.

• You should normally save data to the confocalstor rather than on one of the local hard drives. You can save data to the local hard disks (e.g. if there are problems with the network connection) but the data will only be kept on that local drive for 30 days, after which it will be deleted
• Never save data on the C: drive (this includes the Desktop). Use the D: drive or another large drive if available if you need to save on the local machine. The C: drive is a small system partition that doesn't have the capacity for much data. Filling it up will cause problems
• Defragment both C: and D: partitions before and after time-lapse acquisition. This ensures there are large contiguous sectors of disc available for writing data
• Quit the software and re-open it before and after time-lapse acquisition. This clears the temp folder that contains the temporary image data files for that session