Zeiss LSM510 Meta Confocal Microscope User Guide

More documents

Recommendations

Info

<strong>Confocal</strong> Scanning Find -the system will try to find a preliminary image for you Fast -to continuous scan at fastest speed and w/o averaging - for focusing and gain/offset adjustment Single -to collect one scan - with averaging. Do this before saving. Continuous -to continuous scan with selected scan speed (with averaging) Stop ALWAYS STOP SCANNING New to open a new image window Note: While fast scanning has a short dwell time, the repeated scans can Photobleach as much or more than slow scan! To focus while scanning, use microscope focus knob or use Focus icon under Online Acquisition Note: the nosespiece drifts very badly for first few hours due to temp changes, move knob towards you to correct (this is down!) Viewing Images Image Window (in the middle of ZEN window) Display 2D-Overlay image Split-different channels Range Indicator - click the channels under the image window to switch between ranger indicator and colored images Red - saturated pixels Blue - pixels that are too black (background, or 0s in pixel value) Adjust gain and laser power to minimize saturation Adjust offset to minimize black pixels Crop Zoom Overlay – adds icons to window Scale bar is here. Click on icon and draw on image. Saving images Set speed and averaging and use single to collect an image you want to save Use save icon to the right of image window or File / Save or Save as to your folders on the DATA Drive E: Images is saved in .lsm format (preferred, you can manipulate later with ZEN LE or ImageJ) or tiff/jpeg format as you want.
Z-Stack Z-Stack icon under Multidimensional Acquisition Mark first/last While Fast scanning use focus knob on scope Mark first, mark last Can do this manually or use Stage icon Z-series ALWAYS collects with nosepiece moving UP. If you set up your Z series to start deep in your sample and move down to the coverslip, it won’t. It will go to the end (coverslip) and move up to the start. Focus values get larger as nosepiece moves down. Use optimal interval to set up section thickness or number of slices Change interval / number of slices as desired Before start, check Z-Stack under Multidimensional Acquisition / Information On Experiment Start to start Z-series Viewing Z-Stack Image Window Z Position icon makes scroll bar below image window Gallery is overlay only, check Show Text under Gallery below image window shows the depth from first section 3D allows you view the Z-stack in 3D Auto Brightness Correction When images get dimmer as you go deeper you can brighten them as you collect them Choose first slice, adjust settings, Set A Choose Last slice: re-adjust settings to match (?), Set B The settings that you change will adjust linearly through this series Gain, Offset, Amp Gain, Laser Power Time Series Cycles - choose how many cycles you want to acquire Interval – set time interval between cycles See Actual scan time and dwell time under the Acquisition Mode You can set more than one interval and change in the middle of the series Start – Manual or choose a clock time based on computer clock Stop - Manual, set number of time points Before start, check Time Lapse under Multidimensional Acquisition / Information On Experiment Start to start time series You can save these settings for future use.
Page 1 and 2: Zeiss 510 Meta w/ ZEN Confocal Micr
Page 3: Confocal Mode LSM icon to switch to
Page 7 and 8: Zeiss Axiovert 200 Microscope Specs
Page 9: Set-up for First-time users of the

Zeiss LSM510 Meta Confocal Microscope User Guide

Create successful ePaper yourself

Delete template?

Save as template?