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Introduction
The Imagex-nanoCCD system has all the features of the Imagex-TGi system but has the added dimension of programmable nanosecond timing. Thus, Imagex-nanoCCD can resolve millisecond , microsecond and nanosecond lifetime fluorescent probes. All this from a single imaging system.
As with the Imagex-TGi system The timing features of Imagex nanoCCD are implemented directly on the CCD. Therefore there is no noisy, fragile and highly expensive intensifier to worry about.
The Imagex-nanoCCD Software
The nanoCCD Time-Gating Control allows the user to change Lamp Pulse Rate, Lamp Pulse Width, Gate Delay, Gate Width and Nanosecond Delay directly from the software interface. These parameters are represented by a graphical key which allows you to monitor the relationship between the gating parameters and the lamp repetition period in real time.
Automated nanosecond imaging...
The nanosecond delay controller built into the Imagex-nanoCCD interface can be controlled directly from the Imagex software interface. Varying the nanosecond delay actually changes the time period between the pulsing of the light source (e.g. a nanosecond pulsed laser) and the beginning of the ‘Gate ON’ period applied to the camera. Each of the 16 image banks can be assigned a different nanosecond delay value. The Imagex multi-expose function then allows you to collect a series of nanosecond time-resolved images with a single mouse click...
Generating FLIM Contrast...
Once you have collected a pair or series of nanosecond images you can generate fluorescence lifetime contrast using the inter-image mathematical operations included in the Imagex software...
As well as generating a simple ratio image you can also perform a ‘Transform’ operation. This passes the ratio values through a user-defined ‘look-up table’ to convert ratio data into real-world data such as oxygen tension, fluorescence lifetime or metal ion concentration. Look-up tables used for the Transform operation are generated by the user from real calibration data and offer an extremely versatile method of data processing.
Once you have generated your time-resolved image you can improve it further by using the data-combining capabilities of the Imagex software. A special interface is provided which allows you to weight the FLIM image with the original intensity image to produce a new ‘Intensity/FLIM Matrix’ image which effectively suppresses the noise generated in regions of low signal level whilst maintaining the FLIM contrast and showing how the lifetime data maps onto the original image.
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