Zeiss AxioObserver User Manual Download

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WF-Zeiss_EN_V3 24/01/2017

Switch on the temperature controller ................................................. 2

Switch on the CO2 controller .............................................................. 3

Start the system .................................................................................. 4

Equipment ........................................................................................... 5

Microscope stand presentation .......................................................... 6

Start Zen / Observation with the oculars ............................................ 8

Acquire a widefield image in transmission mode with the color
camera ................................................................................................ 9

Acquire an image (transmission/fluorescence) with the black/white
camera .............................................................................................. 11

Acquire a Z-stack serie ...................................................................... 12

Acquire a time serie .......................................................................... 14

Multiple stage positions .................................................................... 15

Tile scan ............................................................................................ 16

Graphic tools ..................................................................................... 22

How to save your data ...................................................................... 23

Switch off the system ........................................................................ 24

Switch of the temperature and CO2 system ..................................... 25

WIDEFIELD ZEISS AXIO OBSERVER

USER MANUAL

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Summary of Contents for AxioObserver

Page 1: ...e oculars 8 Acquire a widefield image in transmission mode with the color camera 9 Acquire an image transmission fluorescence with the black white camera 11 Acquire a Z stack serie 12 Acquire a time s...

Page 2: ...nce your sample is put on the system and you saved the positions you wanted to acquire to avoid ab important focus shift in z it s best to wait at least 1 hour and re set the focus on your positions 1...

Page 3: ...ockwise direction 2 Turn on the CO2 controller red brick 3 Turn the air compressed bottle valve silver clockwise until you obtain a pressure of 1 Bar in the exit of the pressure regulator right pressu...

Page 4: ...tem 1 Turn on the multi socket power on the shelf to the left of the system The fluorescent lamp is switched on at the same time 2 Switch on the microscope by pushing the START button on the left side...

Page 5: ...chromat 20x 0 8 420650 9901 Plan Apochromatic Dry 0 8 0 17 550 8 2 II 20 1 Sur l tag re On the shelf LD Plan NEOFLUAR 20x 0 4 Ph2 Korr 421351 9970 Plan Neofluar Dry 0 4 0 1 5 7900 0 75 2 II 20 2 Sur l...

Page 6: ...c knob Right side 1 Take the objectives down at the lowest position 2 Take the objective back up to its working position 3 Open Close the fluorescence shutter 4 Filter cube change for fluorescence 2 b...

Page 7: ...ontrast PH or Normanski contrast DIC 7 Aperture Diaphragm opening for the k hler illumination setting 8 Power of the halogen lamp The LED for the fluorescence The fluorescent lamp is composed of 3 LED...

Page 8: ...mission If you want to observe in phase contrast or DIC first select HAL then select PH or DIC on the TFT screen 5 DAPI to observe in blue 6 CFP to observe in cyan 7 GFP to observe in green 8 DsRed to...

Page 9: ...then if you need to the phase contrast or DIC refer to the posters in the microscope room Select BF PH ou DIC on the TFT sreen 1 Select the tab Acquisition 2 Choose TL Camera couleur 3 Open the menu...

Page 10: ...Acquisition 2 Open the menu Acquisition Mode 3 Make a Live 4 Click on Pick a little pipette will appear instead of the mouse arrow 5 Choose an area that is suppose to be completely white in your image...

Page 11: ...ters in the microscope room Select BF PH ou DIC on the TFT screen 1 Select the tab Acquisition 2 Choose Fluo TL Camera N B 3 Open the menu Channels 4 Check the line s with your fluorophores of interes...

Page 12: ...efine only the center of you stack 4 In the menu Z Stack Check the tab First Last 5 Make a Live 6 Set First defines one extremity of you stack and Set Last the other one OR 4 In the menu Z Stack Check...

Page 13: ...ace at the end of the acquisition In the tab Dimensions 13 Z Position allows you to see the different planes of the stack 14 Gallery displays a set of all the images composing the stack 15 Ortho allow...

Page 14: ...o acquire a z stack or tile has to be taken in account in the chosen interval You can measure it by clicking on Measure Speed the value will be shown in the Interval rubric 6 You can minimize the time...

Page 15: ...tion space that will allow you to set your positions 8 Make a Live 9 On the navigation space double click where you want to acquire an image or use the joystick to move the stage 10 Once positioned cl...

Page 16: ...The navigation space is displayed in the center of the software interface Double click where you want to acquire an image or use the joystick to move the stage 9 In the menu Tile Regions click on Tile...

Page 17: ...on Distribute 15 Yellow circles will be distributed in the tile 16 Double click on a yellow circle make a Live set the focus and click on Set Current Z to save it Repeat for each yellow circle OR 17 D...

Page 18: ...g you are adjusting your acquisition parameters you can correct the shadow effects online Find a position without your sample 1 In the channels toolbox 2 select the global correction mode in the list...

Page 19: ...the options section 1 Select Stitching During Acquisition 2 In the section Stage Travel Optimization select Comb 3 Set the Tile Overlap at 20 4 Select Image pyramid during acquisition Thanks to this...

Page 20: ...select the tile you want to stitch 6 Open the menu Parameters 7 Click on New Output 8 Check Fuse Tiles 9 If the shading is not correct click on Correct Shading and select the Automatic mode If you ha...

Page 21: ...ve two different methods to correct the shading To make an automatic correction by computation 9 Check Correct Shading to homogeneize the background 10 Select the mode Automatic for an automatic corre...

Page 22: ...e to the pixel size of the screen 4 Enlarge or reduce the image 5 Show Hide the color channel on the screen 6 See only one channel at a time 7 Show the levels of gray and the saturation In the tab Dis...

Page 23: ...on the image to save 7 Click on the diskette icon 8 Rename tour images 9 Save them in the folder Users on the Data Hardrive All images should be saved in the pathway users year month day your name Sa...

Page 24: ...an the front lens and on the side with optical paper 2 Exit from the software ZEN 3 Transfer your data on your hardrive Check the booking site If there is no one after you 4 Turn off the computer 5 Tu...

Page 25: ...t to break the glass cover 3 Unclip the greenish tub that you will find on the water reservoir on the system 4 Switch off the CO2 controller red brick 5 Turn the valve of the compressed air pressure r...

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