Nikon A1+ Panduan Pengguna - Halaman 22

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can occur if too much light hits the detector.

12. To select a laser line for bleaching press the wavelength button i.e. 405 then set %

power you would like to bleach with. Multiple lasers can be used for bleaching

simultaneously if required.

13. The Scan Speed drop down box refers to the scan speed (frames per second) used

when bleaching the ROI. A slower scan speed will increase the bleaching rate but

may be less suitable if the recovery of the protein of interest is dynamic.

14. Manual Shift Alignment can be configured by facility staff if you find that the bleach

ROI you place on the image doesn't perfectly align with the area actually bleached.

15. Find an area on the sample that you'd like to bleach then select the down

arrow next to the ROI button found in the right margin of the live window.

Select the Simple ROI Editor menu option.

16. Select either the Rectangle or Ellipse ROI tool and draw the ROI onto the image.

The ROI can be resized by moving to the edge of the ROI when the cursor changes

to a double headed arrow. The ROI can be moved around the image when the

cursor changes to a four headed arrow.

17. To Delete a single ROI click on it then press the Delete key on the keyboard

press the Clear button in the Simple ROI Editor tool panel

19. Press Finish when you have finished editing the ROIs

20. Ideally you should have a minimum of 3 ROIs. One assigned as the bleach region

and two others, the first being a Background ROI and the second being a

Reference ROI.

21. Right click on the Bleach ROI and select "Use as stimulation ROI"

22. Ensure that the Background ROI is placed on a black area of the image where there

are no cells. Then right click this ROI and select "Use as background ROI"

23. Place the third ROI on another cell which won't be bleached, then right click on it

and select "use as Reference ROI"

18. To delete all ROIs