PerkinElmer IVIS Lumina III Series, Hardware Manual

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IVIS ® Lumina XRMS Series III Hardware Manual Chapter 8 | Fluorescence Module
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The vertical axis in Figure 8.9 is optical density, defined as OD = -log(T) where T is the transmission.
An optical density of 0 indicates 100% transmission, whereas OD7 indicates a reduction of the
transmission to 1x10
-7
. Typical transmission of a filter in the bandpass region is about 0.7 (OD0.15)
and typical blocking outside of the bandpass region is about OD7. The band gap between the two
filters is usually defined as the gap at 50% transmission (OD0.3). There is a slope in the transition
region from bandpass to blocking, as indicated in
Figure 8.9 . A steep slope is required to avoid
overlap between the two filters.
The fluorescence filters are high quality interference filters, constructed from alternating layers of
dielectric films on a substrate of low auto-fluorescent glass. Care has been taken to minimize filter
auto-fluorescence so that its level is below OD7. Filter passbands for the standard set are listed in
Table 6.2 on page 43 .
8.5 Fluorescent Imaging
IVIS Acquisition Control Panel
Acquiring fluorescent images using Living Image software is controlled through the IVIS
Acquisition Control Panel (
Figure 8.10 ). To acquire a fluorescent image, the user must check the
Fluorescent box on the left side of the panel. Once selected, controls for the illumination lamp –
Fluor Lamp Level, and Filter Lock – will appear in the top half of the panel. Checking the Filter
Lock box ensures that the excitation and emission filters are properly paired. During image
acquisition, the lamp is computer-controlled through Living Image software. The Lamp Level drop-
down list controls the illumination intensity level of the lamp with options – Off, Low, High, and
Inspect. The Low setting is approximately 18% of the High setting. Inspect turns on the illumination
lamp, allowing the user to manually inspect the excitation lamp.
NOTE:
This chapter provides a quick start guide to acquiring fluorescent images. See the Living
Image
®
Software User Manual for IVIS Lumina XRMS Series III for complete details.
Figure 8.10
IVIS Acquisition Control Panel – Living Image Software
NOTE:
Ensure that the correct filters selected from the Excitation Filter and Emission Filter drop-
down lists before you select Inspect. The inspect operation automatically positions the filters that
are selected from the drop-down lists before turning on the lamp. Changing the filter selection in
the drop-down lists will have no effect until another inspect operation is performed.