Olympus Confocal Microscope奥林巴斯共聚焦显微镜.doc

Olympus Confocal Microscope:Instructional GuideIf you are the first user of the day, turn on the instrument; the last user of the day turns off everything, except the computer.Open the fluoview program: FV10-ASWWait until all windows are open (this should be automatic if everything is turned on) Image Acquisition Control:Dye List: To select dyes, select the dye-list icon. The maximum dyes per phase are three, any more and the virtual channel scan must be selected. If two dyes are too close to each other in wavelength, then they must be separated in two different phases. HV: This controls the PMT levels, so if it is increased, the signal received by the detector of that dye would be amplified and vis versa. Do not set above 800 or it will become non-linear, analogous to using gain.Gain: When gain is increased, the strongest signals are amplified and not the weakest. This is not good for later analysis of data. Do not use gain unless absolutely necessary to see signal. Offset: When offset is increased, the background noise will disappear. Try not to abuse this function or real signal can be masked. To use properly, select a high-low setting from the LUT menu, located under the Live View window (looks like a rainbow). OR you can press “Ctrl-H” and it will toggle all of the channels together in high-low. To escape this setting, press “Ctrl-H” again. When in high-low, whatever looks red is over saturated pixels. Decrease the HV or laser levels until the signal is strong but not overpowering. When offset is increased about 0, you will start to see a blue background. Increase only until the background is solid blue and stop there.Epi Lamp: Press this button to turn on or off the epifluorescent lamp, allowing you to visualize your sample via the ocular piece. Focus x2: This is the most common button used to initiate the lasers, projecting an image in the Live View window. It will drop the acquisition time down to 2?s/pixel if AutoHV is selected (should always be on).