Imaging systems for luminescence
Current systems, established by Andrew Millar's lab in Edinburgh (2005).
The workhorse for our current luminescence imaging is Hamamatsu's Orca II BT 1024 monochrome camera (model number C4742-98-26-KWG2). 16 bits depth and -75ºC Peltier cooling make this camera suitable for luminescence signals without binning. We control this camera using Hamamatsu's low-cost Wasabi software via a Firewire interface.
Previous systems, established by Andrew Millar's lab in Warwick (1996-2004)
One system comprised a Hamamatsu Orca II monochrome camera to capture fluorescence images. 14 bits depth,-50C Peltier cooling and 2x2 binning made this camera suitable for bright luminescence signals. It was controlled by MetaMorph via a Mutech MV-1500 framegrabber.
The other type of low-light system used a SITe 512x512 back-thinned CCD in a liquid-nitrogen cooled camera head from Princeton Instruments (a division of Roper Scientific) 16 bits depth and -120ºC operating temperature, controlled by MetaMorph software from Universal Imaging Corp. This camera looks identical to the one used in Photometrics ChemiPro system, which is also used for luminescence imaging by Jian-Kang Zhu's lab in Arizona, as described in this profile. Our system included a modified, 100L liquid nitrogen autofill system from Kadel (which caused us significant headaches), a home-built dark chamber and a home-built remote focus motor. Additional camera control and image analysis software was purchased from the camera manufacturers.
Our motorised microscopes were from Zeiss (Axiovert 135TV and Axioplan2). The key components are high-transmission Fluar objectives and "direct" imaging ports that mimimize the number of glass elements between sample and CCD.
Earliest systems, established in Warwick and in earlier posts.
We have previously used a two-stage micro-channel-plate intensified CCD (model C2400-47) from Hamamatsu Photonics, under the control of Hamamatsu Argus50 software. Hamamatsu sells a dark imaging chamber with this system. This was the major type of camera used in the Chua (NUS and Rockefeller) and Kay (U.Va.) laboratories during our original characterisation of the luciferase reporter (1989-1994) and was used in Warwick from 1996-2003. The technology is reliable but the detector is not robust to high light, making it risky to use with automated lighting systems.
See also Luciferase for other labs using low-light cameras to detect luciferase.
Video Imaging of Regulated Firefly Luciferase Activity in Transgenic Plants (minireview).
