Zeiss LSM 880 Manual de usuario

880 Table of Contents

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

Setting 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 Light Image……………………………..18

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

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

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

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

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.

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 – Touch “Home”

then “Microscope” and click on the “Objectives”

Tab.

3. After you choose your objective make sure to

clean it with lens cleaner

4. Set your sample on the stage, please remember

to add oil or water as appropriate

Viewing your sample

1. To view your sample through the eyepiece

select what color you would like to use.

If you would 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 objective down

Away from yourself moves the objective up

3. Use the joystick next to the microscope to move your sample.

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. Please contact Jeff, Agnes or Erica if you need to set up a new configuration.

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. 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.

Please talk to Jeff, Agnes or Erica if you have

questions about selecting the appropriate level

for the laser intensity.

4. Once you have your image in focus and at the intensity you like, click the

Snap button to acquire your data quality scan. 8

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

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.

Note: Examples of commonly used dyes and the

laser lines used to excite them are listed on the

next page.

Note: The files that start 880 are for the regular

confocal and the ones that start AS are for the

Airyscan

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