Olympus FLUOVIEW FV300, User Manual

Page: 294 / 564

APPLIED OPERATIONS
/ Protocol processor
IV.
OPERATION INSTRUCTIONS
IV .
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Set each Track as sown below;
z
Setting Track 1
In XYT observation, set the zoom ratio to 2X and acquire an image before bleaching.
1. Select [New mode]-“XYT” in [Mode] of [Track].
2. Select “Normal” in [Start by] of [Track].
3. Enter “1” in [Condition] of [Track].
4. Enter “0” to [Frame Interval].
5. Enter “2” to [Zoom].
6. Select “Middle” in [Scan Speed].
7. Enter the laser intensity value for image acquisition to [Laser Intensity]
8. Enter the PMT value of each channel to [PMT Voltage].
9. Enter a comment in [Comment] if necessary.
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Setting Track2
In XYT observation, set the zoom ratio to 10X and acquire three images. (Irradiate strong
laser onto the region to bleach.)
1. Select “Series mode” in [Mode] of [Track].
2. Select “Normal” in [Start by] of [Track].
3. Enter “3” to [Condition] of [Track].
4. Enter “0” to [Frame Interval].
5. Enter “10” to [Zoom].
6. Select “Slow” in [Scan Speed] for fast photo-bleaching.
7. Enter the laser intensity value for image acquisition to [Laser Intensity]
8. Enter the PMT value of each channel to [PMT Voltage].
9. Enter a comment in [Comment] if necessary
To properly irradiate the laser for bleaching, enter a larger value in
[Condition] and fast forward the tracks using the <Skip> button in
accordance with the grade of bleaching.
z
Setting Track 3
In XYT observation, return the zoom ratio to 2X and acquire 30 images every 5 seconds.
(Observe the change in fluorescence image with time after bleaching.)
<Skip> button
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