Tutorial Guide #3 – Taking a Z Stack (Example 1)

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1. STARTUP AND SET INITIAL LASER POWER

Step 1: Wait for the Startup Screen

Step 2. Turn Scanning ON

Step 3. Set an Initial Laser Power, and Enter Settings Menu

2. SET DESIRED IMAGING MODES

Step 4: Choose Setting Button to and Enter Settings Menu

Step 5: Set the Desired Aspect Ratio (i.e. 1:1)

Step 6: Set the Desired Speed/Resolution (i.e.

Step 7: Choose the Appropriate Filter for the sample and dye used

3. IDENTIFY Z STACK REGION OF INTEREST

Step 8: Find the desired starting position for the Z Region you want to image

Step 9: Find the desired ending position for the Z Region you want to image

4. IDENTIFY Z STACK AREA AND ADJUST LASER POWER

Step 10: At Start Position. Look At picture and Brightness Value. Too HIGH

Step 11: Increase Laser Power at Start Pos

Step 12: Scan other regions of Z – If Brightness too High Reduce Laser Power

Step 13: Laser Power Too High at this depth – Reduce so Brightness increases

5. ENTER Z STACK MENU AND SET START POSITION

Step 14: Go to Start Position

Step 15: Enter Z Stack Menu

Step 16: Go to Start Position

Step 17: Set Start Position

6. LOOK FOR LOWEST BRIGHTNESS VALUE THROUGH Z REGION

Step 18: At Position (-14um) Brightness = 22%

Step 19: At Position (-6um) Brightness = 7% (LOWEST IN Z REGION)

Step 20: At Position (+7um) Brightness = 15%

Step 21: At Position (+27um) Brightness = 28%

7. SET END POSITION AND ADJUST BRIGHTNESS MANUALLY TO LOWEST VALUE IN REGION

Step 22: Set the End Stop Position

Step 23: Set Increment (i.e. 3um)

Step 24: Turn Auto Brightness to OFF

Step 25: Change Brightness to Lowest Value (i.e. 8%)

8. CHOOSE Z STACK METHOD AND PRESS START

Step 26: Choose Z Stack Method

Step 27: Press Start

9. Z STACK IN OPERATION

Step 28: Z Stack has started goes to start position

Step 29: Z Moves towards end position at increments taking image captures

Step 30: Z Moves towards end position at increments taking image captures

Step 31: Z Stack Operation Completed

10. THE RESULTING Z STACK IMAGES

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Manufactured by

Optiscan Pty. Ltd. Head Office

16 Miles Street, Mulgrave

Victoria 3170, AUSTRALIA

Phone: +61 3 9538 3333

Fax: +61 3 9562 7742

www.optiscan.com

Sales

sales@optiscan.com

Customer Support

service@optiscan.com

Additional Resources and Links

www.optiscan.com

www.viewnvivo.com