Invitrogen EVOS M5000 - Image Files

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Sync Z: The Sync Z feature is designed to enable the user to focus on a single channel while the
other channels are adjusted automatically. This is accomplished by first adjusting the focus of each
channel individually with the Sync Z feature unchecked. This will establish unique osets for each
channel. Once this initial adjustment is complete, the Sync Z feature can be enabled and thereafter
focusing on one channel will automatically adjust the focus of the others using the same osets
that were established during the individual setup. If at any point the osets become misaligned, the
Clear Z Osets button can be used to quickly reset and start again.
Image files
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Channel: A channel refers to a specific illumination range used to capture images. Fluorescence
channels are defined by their excitation and emission wavelengths, which enable the detection of
dierent cell populations or subcellular structures. Transmitted light is full-spectrum and can be
used either alone or in conjunction with phase-contrast optics to provide images based on the light
that passes through the sample. The interaction of the light with the sample as it passes through
cells or tissue provides contrast for transmitted light images.
•
Channel overlay: When multiple channels are captured from a single location, they can be overlaid
in an image to visualize the relationships between dierent cells or structures identified by each
channel. The overlaid channels are pseudocolored, meaning that distinct colors are assigned to
each channel for better dierentiation. It is important to note that an image with overlaid channels is
intended for display purposes only and should not be used for quantitative analysis.
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Analysis image: An analysis image refers to a single-channel image that is saved in a lossless
format using the full dynamic range of the camera (12 bits). These images are saved in 16-bit
format, which may require specialized viewing software for proper display. When viewed on other
software, analysis images are displayed in grayscale monochrome, representing the pure data
that was captured. To enhance visualization, the EVOS
™
software automatically displays analysis
images using the appropriate pseudocolor. It's important to note that the EVOS
™
software does not
modify analysis images in any way. This means that features like scalebars or annotations cannot
be added, and changing the contrast onscreen does not alter the underlying data in the image file.
•
Display image: A display image refers to a modified version of an original image that can be
used outside of the EVOS
™
M5000 software. These images are usually pseudocolored, either
representing a single channel or multiple overlaid channels, and are intended for purposes such as
PowerPoint
™
presentations or sharing with colleagues. It's important to note that display images
are saved in an 8-bits per channel file format, which may compromise the intensity information
and result in lower quality images for quantitative analysis. While display images can be useful
for manual tasks like cell counting or size/distance measurements, it is not recommended to
use them with analysis tools like Auto Count, Confluence, or Transfection Eciency, as these
measurements rely on the complete intensity information that might be lost in display images.
Display images can be customized with features like scale bars or annotations, and adjusted
brightness and contrast values for visual presentation; as such they are not the original data and
should not be used for analysis purposes.
AppendixEGlossary
Image files
E
EVOS
™
M5000 Imaging System User Guide
143