Choosing a software for acquiring two colour images with two cameras.
When acquiring two color images with two cameras on TuCam it is essential that the acquisition software is compatible with the detectors used and is capable of acquiring simultaneous image capture from both cameras. This section provides information concerning compatibility of Andor’s range of imaging cameras for microscopy with a number of acquisition softwares, covering Andor Solis, Andor iQ, MetaMorph from Molecular Devices Corporation and NIS-Elements from Nikon Instruments Inc.
The following table summarizes the software packages that are compatible with Andor’s cameras when operated under simultaneous dual camera acquisition.
Instances of S/W required
2 x Neo sCMOS
2 x iXon Ultra
2 x iXon 885
2 x iXon 897
2 x LucaR
2 x Clara
* Only the 2x Neo camera need to be operated under two instances of NIS Elements. All other dual cameras indicated can be run under one instance.
As conveyed, two instances of the imaging software Andor Solis and Andor iQ are required to run Andor’s range of imaging cameras as these software packages do not have the dual camera drivers available, which are required to capture simultaneous images from two cameras in one software window. This implies that when using the TuCam with two cameras from the Andor range, each camera will require its own instance of Andor iQ and Andor Solis to achieve simultaneous and synchronous capture.
NIS-Elements and MetaMorph acquisition software packages have dual camera drivers available for the majority of Andor’s range of imaging cameras. This indicates that only one instance of the software is required to acquire images simultaneously from both cameras in one window. However, there is one exception in NIS-Elements and the Neo sCMOS. NIS-Elements do not have a dual device driver for the Neo sCMOS and hence two instances of NIS-Elements are required in order to simultaneously acquire with two Neo cameras. For dual wavelength imaging, the Master-Slave set-up can be used which is explained in the technical note entitled ‘Dual Wavelength Imaging using TuCam: Software Set-up Focus’. This set-up also applies to acquiring two color images simultaneously when using Andor Solis and Andor iQ.
Merging simultaneous two color images following acquisition with TuCam.
Following acquisition of dual wavelength images with Andor iQ software it is possible to merge these images by bringing both acquired images into one instance of iQ and then merging the two files under the Process Menu-Dual image disk. Assuming the meta data from both files is identical this will be possible i.e. the x,y,z & t dimensions. If Andor Solis is the acquisition software of choice it is possible to open the Solis files in Image J and use the merge tool here. Merging and analysis functionality also exists within MetaMorph and NIS Elements packages.
Merging simultaneous images acquired using the Optosplit II
Following acquisition with Optosplit II, analysis plug-ins are available in iQ, MetaMorph, MetaFluor, Image J and NIS-Elements for image registration and dual wavelength image merging.