Zeiss LSM 510 META Manual de usuario descargar pdf (Pagina 10)

Z-sectioning

To collect a series of images at different points along the z-axis the settings can be inputted in two

ways: Z Sectioning or Mark First/Last.

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Z-sectioning - takes x slices at a particular z interval

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Mark first/last - set the top and bottom of your sample and select either the interval or number of

slices (the other is calculated for you)

Both of the methods described here assume that the user has optimized and set all imaging

parameters, including pinhole size, detector gain/offset, scan time and averaging described above.

Mark First/Last Method

1. Select Z Stack at the top of the "Scan Control" window - doing this will activate the Z Settings panel.

2. Determine the start and end points of the Z stack. (Z sectioning is performed moving into the

specimen, away from the coverslip.) To find the starting point for the Z stack perform a Fast XY scan

and focus toward the coverslip (rotate fine focus knob toward you) until you reach the point at which

you'd like to begin the Z stack. Stop the Fast XY scanning.

3. Click Mark First in the Mark First/Last tab in the Z Settings panel.

4.Start another Fast XY scan and turn the fine focus knob away from you focusing down through your

specimen until you reach the point at which you'd like to end the Z stack. Stop the Fast XY scanning.

5. Click Mark Last to set the end point of the stack.

6. Assuming you have already adjusted your pinholes as described in "Acquire Menu: Collect An XY

Section", look up the optimal z interval thickness by clicking Z Slice at the top of the Z Settings

box. This will open the "Optimal Slice" window.

7. Optimal z interval thickness for the objective currently in use will be displayed in this

window. Clicking Optimal Interval in the lower left corner of the window will set the z interval of your z

stack to the recommended thickness.

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Zeiss LSM 510 META Manual de usuario Pagina 10