Dwell Time
per Pixel (P:)
Box Size
Scan Time
Per Line (L:)
Seconds
per Frame (F:)
Time for Set
(S:)
2.0
µ
s
512X512
1.572 ms
0.822 s
2.446 s (Sequential)
2.0
µ
s
512X512
1.572 ms
0.822 s
0.822 (Simultaneous)
8.0
µ
s
512X512
6.288 ms
3.288 s
9.862 s (Sequential)
8.0
µ
s
512X512
6.288 ms
3.288 s
3.288 s (Simultaneous)
4.0
µ
s
1024X1024
6.288 ms
3.288 s
9.862 s (Sequential)
4.0
µ
s
1024X1024
6.288 ms
3.288 s
3.288 s (Simultaneous)
Table 3. Effect of Scan Speed and Box Size on Acquisition Time - single plane, 3-channel image
Scan Time per Line (L) is determined by Dwell Time (P) X length (e.g. 512, 1024, etc.) plus the
“flyback” time required to return the laser spot to the start of the next line. The time to record a Frame
(one image, F) is L X number of lines in the box size. The Set (S) is F X the number of scans required
to record each 2D image plane. S = F for simultaneous collection, but S = F X # channels for sequential
acquisition. Kalman averaging will increase scan time by the number of averages, and a z-series will
increase S by the number of focus steps (F X z-steps).
2.6.3 Scan Regions, Figure 6.3.
These buttons select the type of region to scan.
The leftmost “Normal” rectangle defines the scanned region as the selected box size. The Polygon
options allows user-defined regions within the box. These may be rectangular, circular or irregular
polygons. The laser beam is blanked outside of the selected scan region.
The ‘Line’ and ‘Point’ options are specialty scans that may be used for creating a “vertical” section by
scanning through a focus range or to record rapid dynamic processes, such as calcium spikes. All of the
scan regions may be used for techniques such as photoablation, photoactivation or photobleaching.
2.6.4 Image Aspect Ratio, Figure 6.4.
The default scan region is 1:1 (width:height), or
square
. The “
4:3
” option provides rectangular boxes,
e.g. 768 X 512. The “
arbitrary
” option allows the user to define any needed box, e.g 129 X 17.
2.6.5 Box Size, 6.5.
Box Size controls the “sampling density” of the image, the number of sampling intervals in the X-axis
and Y-axis of the scanned field. The scanned region will be divided into the stated number of lines with
each line divided into the selected number of sample points. In turn, the acquired image will possess
corresponding dimensions, in pixels.
Olympus Fluoview-1000 User’s Guide
V.M. Bloedel Hearing Research Center, Core for Communication Research
Center on Human Development and Disability, Digital Microscopy Center
May 11, 2011
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