Zeiss LSM 880 User Manual

Inverted confocal microscope with airyscan

Hide thumbs Also See for LSM 880:

Operating manual (690 pages)

Notes on device safety and installation requirements (50 pages)

Basic user's manual (25 pages)

880 Table of Contents

880 User guide overview.........................................................2-9

Setting up configurations on your own.................................10-11

Acquisition mode- Recommended settings..........................12

Tile Scans and Stitching............................................................13-14

Time Series.................................................................................15

Z-Stacks.......................................................................................16-17

Adding a DIC Transmitted Light Image...................................18

Kohler Illumination....................................................................19

Incubation...................................................................................20-21

Calibration for Airyscan.............................................................22-25

Table of Contents

Need help?

Do you have a question about the LSM 880 and is the answer not in the manual?

Questions and answers

Summary of Contents for Zeiss LSM 880

Page 1: Table Of Contents
880 Table of Contents 880 User guide overview………………………………………………...2-9 Setting up configurations on your own…………………………...10-11 Acquisition mode- Recommended settings………………..12 Tile Scans and Stitching…………………………………………………...13-14 Time Series………………………………………………………………………15 Z-Stacks……………………………………………………………………………16-17 Adding a DIC Transmitted Light Image……………………………..18 Kohler Illumination…………………………………………………………..19 Incubation………………………………………………………………………..20-21 Calibration for Airyscan…………………………………………………….22-25...
Page 2: 880 User Guide Overview
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...
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...
Page 4 Turning 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 warm up.
Page 5 Viewing your sample 1. To view your sample through the eyepiece click the locate tab 2. Choose your objective by clicking on the objective button, this will bring up a list of objectives currently on the microscope. You may also choose your objective by using the touch screen next to the monitor –...
Page 6 Viewing your sample 1. To view your sample through the eyepiece select what color you would like to use. Blue Green Red If you would like to see white light select DIC 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.
Page 7 Setting up a configuration 1. To load a configuration from a 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.
Page 8 Taking an image 1. To view your sample click on the Live button. 2. You may need to adjust to focus by using the focus wheels on the microscope or on the side of the touch screen. 3. To change the intensity of your sample go to the channels tab and click on the track you want to adjust.
Page 9 Saving your file 1. After 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, under images and documents...
Page 10: Setting Up Configurations On Your Own
Setting up configurations on your own 1. Click on button that looks like an open file folder it will bring up a menu called Load Acquisition Parameters 2. Choose what laser lines you would like to use to excite your sample. Then click on the setup that contains those lasers.
Page 11 What laser should I use? Here are some examples of commonly used dyes and the laser lines used to excite them: Laser Dapi, Alexa 405 GFP, FITC, Alexa 488 Alexa 543, Alexa 568, Alexa 594, Rhodamine, Mitotracker red Alexa 594 CY5, Alexa 633, Alexa 647...
Page 12: Acquisition Mode- Recommended Settings
Acquisition mode – Recommended settings 1. A good Frame Size to use is 512x 512, if you need to change your Frame Size click on the X*Y button 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.
Page 13: Tile Scans And Stitching
Tile Scans 1. To do a tile scan, first click Tile Scan, a Tile Scan box should appear on the right if it doesn’t scroll 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.
Page 14 Stitching a Tile Scan 1. After you take your image save your file - click on File then Save or - click on the disk icon on the top left 2. Click on the Processing tab 3. Under Method click on Stitch 4.
Page 15: Time Series
Time Series 1. To do a time series, first click on Time Series. 2. The Time Series tab 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.
Page 16: Z-Stacks
Z-Stacks 1. To make a z-stack first click on Z-Stack, a Z-Stack box should appear on the right if it doesn’t scroll down on the left until you see it. 2. If you would like to move the Z-stack box, click the undock button 3.
Page 17 Z-Stacks using Center 1. To make a z-stack first click on Z-Stack, a Z-Stack box should appear on the right if it doesn’t scroll down on the left until you see it. 2. If you would like to move the Z-stack box, click the undock button 3.
Page 18: Adding A Dic Transmitted Light Image
Adding a DIC Transmitted Light Image 1. If you would like to add a 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 check out the Kohler illumination cheat sheet on the next page.
Page 19: Kohler Illumination
Kohler Illumination for the 880 1. Bring the sample into focus 2. Close 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.
Page 20: Incubation
Incubation 1. Before you plan to use incubation please talk with Jeff, Agnes or Erica to help with setup 2. Make sure to turn on the incubation and CO2 controls before you open Zen (there is one switch on the TempModuleS and one on the Heating Unit XL S) Note: The TempModuleS light will turn 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.
Page 21 Incubation 5. Type in the temperature you would like to use for your incubation, for either the insert, or the incubation chamber 6. The numbers on the right hand side are the actual temperature for the insert and the incubation chamber 7.
Page 22: Calibration For Airyscan
Calibration for Airyscan When you are setting up your calibration 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. Remember in Airyscan 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 provided by the FMIC to do your calibration.
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...
Page 24 Calibration for Airyscan 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.
Page 25: Calibration For Airyscan
Calibration for Airyscan 8. In the Airyscan box, look at the Quality and Status. It will begin with “Unknown (waiting)” in white, then go to “Bad (Waiting)” or “Bad (adjusting)” in red and when it is completed it will say “Good (Waiting)”...

Zeiss LSM 880 User Manual

Table of Contents

880 Table of Contents

Need help?

Questions and answers

Related Manuals for Zeiss LSM 880

Summary of Contents for Zeiss LSM 880

Table of Contents