Olympus Fluoview-1000 Gebruikershandleiding - Pagina 45

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V.M. Bloedel Hearing Research Center, Core for Communication Research

Center on Human Development and Disability, Digital Microscopy Center

Figure 15. The Stimulus Setting window for photoactivation.

1. Select a field in the sample that contains 1 or more regions of interest;

2. Collect a scan of this field with the desired acquisition settings for each channel;

3. Set the Stimulus window settings as shown in Figure 15;

a. 'Use Scanner' - choose 'Main', we don't have a SIM scanner

b. choose the ROI type and mark one or more ROIs on the Live Display window

i. each bleach ROI will have "B" appended to the ROI number

c. set the pixel dwell time for bleach scanning;

d. 'Laser' - select one or more laser lines for bleaching, and their power level

e. 'Stimulate Start Setting'

i. 'Manual' requires initiating scanning with the XY button and stopping with Scan Stop button

ii. 'Auto Stop' halts bleaching after a given number of scans or a defined time period (ms)

f. 'Activation in Series' - allows photoactivation operations in a sequence

i. 'PreActivation' - record a specified number of images before activation

ii. 'Activation Time' - Set number of scans or a defined time period (ms) for activation

iii.a set of images after bleaching can be collected

4. The time interval between scans and number of total scans (including pre-bleach and bleach scans) is

set by the Timecourse window;

May 11, 2011

Olympus Fluoview-1000 User's Guide

Select scanner

Choose entire field or ROI selections

Set dwell time

Choose laser line and power level for

bleach

Start bleach manually

Stop bleach manually or automatically after

n frames or n ms [optional]

Bleach in a series of images

Set the number of initial frames

Set number of bleach frames or bleach time