UVP iBox Scientia Small Animal Imaging System - Manual (Page 13)

iBox Scientia Small Animal Imaging System 13
11. After adjusting the lift, ensure that the Saturation preview
box is checked, adjust the focus and the exposure time so
that the preview is not saturated (saturated areas appear
red when the Saturation preview box is checked).
12. Capture the mouse image. The captured image can be
saved to be used later in the Composite Image step.
Capture Fluorescent Image of Mouse/Rat
1. Select the appropriate BioLite filter and the Darkroom filter
from that corresponds to the fluorescent protein in the
mouse. (For example, if the mouse has a GFP fluorescent
target, use the GFP BioLite filter and GFP Darkroom filter.)
2. Change the Exposure time to 1 second.
3. Ensure that 4X4 binning is selected for preview and that
1X1 binning is selected for capture.
4. Ensure that Compensate exposure for binning box is
checked.
5. Capture the image. The captured image can be saved to
be used later in the Composite Image step
If the target is saturated (is showing red or yellow areas),
reduce the exposure time to half [1sec->500msec-
>250sec…].
If the captured image shows a flat dark image, increase the
exposure time in steps of 20 seconds [1sec->20sec-
>40sec->1min…] until yellow areas appear. Once yellow
appears reduce by 20 seconds.
If the captured image shows a bright image looking no
different from the white light image, the fluorescent signal
might not be strong enough to be observed. (Fluorescent
proteins can take 48 hours to express. Implanted, in-depth,
fluorescent tissue can take one week to grow to a
discernible size. In these two cases, one might just have to
wait another day to image again.)
Create a Composite Image of a Mouse/Rat
1. Open the Image Action Tab and click onto the Multi-Image Actions Menu Button.
2. The Image Compositing window appears.
3. Select the number of images to make the composite. One RFP tumor, for example, will need two images
to create the composite.—a white light (or visible) image and a fluorescent image. (A Pseudocolor may
be added to the fluorescent image to create a colorful Pseudocolor map as shown below.)
4. Use the drop down menu to open the saved image of the white light mouse or rat created using page 16
instructions for capturing a white light image.
5. Use the drop down menu to open the saved image of the florescent mouse or rat created using page 17
instructions for capturing a fluorescent image.
6. Select the Mode desired for your application. (Overlay is more pixilated, whereas Blend is smoothed out)
7. Select the Pseudocolor map checkbox to view the legend of the colors depicted in the image.
8. Select the desired florescent vs. white light combination by moving the Threshold slider for the image.