Photometrics Evolve 512, User Manual

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26 Evolve 512 Users Manual
Evolve 512
Application
Examples
Example 1:
“I would like to obtain kinetic information (time-correlated) information of the single molecule
fluorescence. My light level is fairly low and I want to optimize the frame rate of the camera.”
For this application, the "Multiplication" port (Port #1) should be selected and camera speed
should be set to 10 MHz. In addition, the camera should be operated in “Overlap mode”.
This can be achieved by setting the camera to “frame transfer mode” with clearing mode
“Pre-sequence” and number of clears “1” (refer to Figure 8). If possible, choose a sub-region
(ROI) and/or binning to further increase the frame rate. Finally, on-chip multiplication gain
can be used to boost the signal level and achieve a high signal-to-noise ratio.
Example 2:
“My application requires precise control of the exposure time (less than the readout time) and I want
to operate the camera in the most sensitive mode possible.”
For better imaging characteristics, the "Multiplication" port (Port #1) is selected and camera
speed must be set to 5 MHz. Use on-chip multiplication gain for increased sensitivity. To
achieve an exposure time less than readout time, use Non-Overlap mode (shown in Figure
6) and set the clearing mode to “Pre-exposure” and clear count to “2”.
Example 3:
“I would like to image a bright target/sample (e.g., fluorescence or bright field) that is fixed. High
resolution and high dynamic range are important."
For imaging bright, non-moving samples, the "Non-multiplication" port (Port #2) offers the
best performance in terms of dynamic range. Set the speed to 1.25 MHz (or 5 MHz, if higher
speed is required) and conversion gain to #2 (this allows maximum full well of the CCD to
be used). Reading out the full CCD (no-ROI or binning) gives best resolution possible. In
addition, camera can be operated in overlap or non-overlap mode as explained before.
Application Settings
Summary
The table below summarizes typical Evolve 512 settings for two types of applications and is provided
as a setup aid. Actual settings will vary based on the exact nature of your experiment design.
Application
Readout
Amplifier
(Port)
Readout
Speed
Multiplication
Gain
Conversion
Gain
(e-/ADU) Readout Mode
Kinetic samples
(single molecule
fluorescence or
tracking) Multiplication
(Port #1) 10 MHz or
5 MHz Use as required to
obtain best signal-
to-noise ratio
(SNR).
Typical
EM gain values
are around 200.
#2: 6e
-
/ADU Overlap mode
(Frame Transfer,
Pre-Sequence)
Fixed samples
(bright field) Non-
multiplication
(Port #2) 1.25 MHz Not Applicable #1: 4e
-
/ADU Overlap or Non-
overlap
Table 2. Application Type and Evolve 512 Setup Parameters