Zeiss LSM 880, User Manual

Page 1: ...ing up configurations on your own 10 11 Time Series 15 Z Stacks 16 17 Calibration for Airyscan 22 25 Acquisition mode Recommended settings 12 Tile Scans and Stitching 13 14 Adding a DIC Transmitted Li...

Page 2: ...880 User Guide Overview Starting up the system Turning on the lasers Viewing your sample Setting up a configuration Taking an image Saving your file 2...

Page 3: ...Starting up the system 1 Follow the directions for Powering up the 880 this is located on the wall behind the 880 monitor 2 Double click on the Zen icon 3 Click on the Start System tab 3...

Page 4: ...on the lasers 1 The system starts with the locate tab open 2 To turn on the lasers click the acquisition tab 3 Use the pulldown tab to turn on the lasers Note the 488 laser takes about 5 minutes to wa...

Page 5: ...ring up a list of objectives currently on the microscope You may also choose your objective by using the touch screen next to the monitor Touch Home then Microscope and click on the Objectives Tab 3 A...

Page 6: ...like to see white light select DIC Blue Green Red 2 Use the focus wheels on the side of the Microscope or on the side of the touch screen to bring your sample in focus Towards yourself moves the objec...

Page 7: ...previous experiment go to file open then choose the experiment you would like to reuse 2 Click the reuse button either on the top left of the screen or at the bottom of your image 3 Please contact Jef...

Page 8: ...click on the track you want to adjust You can increase or decrease your intensity by adjusting the Gain Master It is recommended you stay between 600 800 4 You may also need to adjust the laser power...

Page 9: ...you have taken your image you may save your file by using one of the following options clicking on File then Save clicking on the disk icon on the top left clicking on the disk icon on the right unde...

Page 10: ...ameters 2 Choose what laser lines you would like to use to excite your sample Then click on the setup that contains those lasers Note Examples of commonly used dyes and the laser lines used to excite...

Page 11: ...mples of commonly used dyes and the laser lines used to excite them Laser Dye 405 Dapi Alexa 405 458 CFP 488 GFP FITC Alexa 488 514 YFP 561 Alexa 543 Alexa 568 Alexa 594 Rhodamine Mitotracker red 594...

Page 12: ...ton 2 Speed should be set about 7 if you just want to do a quick scan you can increase the speed by clicking on Max 3 For Averaging 2 or 4 are a good numbers to use 4 Bit Depth should be 8 bit 5 Direc...

Page 13: ...oll down on the left until you see it listed under Multidimensional Acquisition 2 If you would like to move the tile scan box click the undock button 3 Under the Centered grid tab choose the number of...

Page 14: ...he disk icon 2 Click on the Processing tab 3 Under Method click on Stitch 4 Click on the image you would like to stitch and then under Method Parameters click on Select 5 Next click on Apply this will...

Page 15: ...will appear under the Multidimensional Acquisitions on the left side To move the Time Series box click the undock tab 3 Set the number of Cycles for how many images you would like to take 4 Set the I...

Page 16: ...box click the undock button 3 Click on live 4 Find the bottom of the sample roll the focus wheel towards your body then click Set First 5 Find the top of the sample roll the focus wheel away from you...

Page 17: ...the left until you see it 2 If you would like to move the Z stack box click 17 the undock button 3 Click on live would like to image Click on the C to take a Snap of the center image to make sure you...

Page 18: ...DIC transmitted light image click the T PMT while you are under the Acquisition tab 2 To have the best DIC image check your Kohler illumination if you need help with this ask Jeff Agnes or Erica or c...

Page 19: ...the field diaphragm until you can see at least one edge 3 Adjust the condenser height until the Edges of the diaphragm image are crisp 4 Center the diaphragm image using the two centering screws 5 Ope...

Page 20: ...rn on when you switch it on the light on the Heating Unit XLS will not turn on until the temperature is turned on in Zen 3 Under the Locate tab you will find the Incubation tab 4 Check the H Insert P...

Page 21: ...the incubation chamber 7 After you have completed your experiment uncheck the H Insert P and H Unit XL and set the temperature to zero NOTE After you have unchecked the H Unit XL make sure the lights...

Page 22: ...you should not adjust the pinhole or the gain you should only adjust the laser power Note If you feel your sample is not bright enough or if you are afraid it may get bleached out you may use a slide...

Page 23: ...Calibration for Airyscan 3 Open Airyscan undock and click adjust in continuous 4 Click on the Acquisitions tab 5 Change the frame size to 512 x 512 23...

Page 24: ...ryscan 6 Highlight only one channel and choose a zoom of 5 Remember it is best to use a channel with bright fluorescence one that will fill most of the scanned area and is in the visible range 7 Click...

Page 25: ...in red and when it is completed it will say Good Waiting in Green Once your calibration is complete uncheck the Adjust in continuous scans Note if it stays in Unknown waiting in white for a long time...