The recent developments in cell biology using bioimaging techniques are constantly bringing new insights into the importance of understanding the mechanisms that govern living cells. The MetaMorph® Imaging System integrates software and hardware and enables researchers to automate image acquisition, analysis and processing from digital CCD cameras.
MetaMorph®’s acquisition tools allow the user to perform a variety of functions, namely to control the exposure time; select a subregion for viewing; control the binning mode gain levels and other parameters specific to the camera type and model. In addition to the X and Y dimensions, it allows the user to construct Z-series, and to use multiple wavelengths and multiple sites with a motorized stage. Once the images are captured, the processing tools enable the researcher to use extensive analytical features to measure distances, areas or fluorescence intensities, brightness and contrast, to count cells, create 3D reconstructions, combine multi-stacks of fluorescence images and create time lapse movies.
I have used MetaMorph® System Version 5.0 to analyze living and fixed material, namely mammalian oocytes. Our system is incorporated into a PC-compatible 32-bit computer with a 2 GHz Intel Pentium 4 processor and Microsoft® Windows® XP operating system. The computer is attached to an IM-35 Zeiss Inverted Microscope interfaced with a Hamamatsu Orca digital camera. The MetaMorph® System provides a high control of all the components of the microscope associated with image acquisition, like the filter wheel, shutter and the motorized stage, and also with the digital camera, focus motors, and all the other motor devices.
When analyzing living material, images of living cells are acquired and compiled to generate time lapse movies. Although conditions of light, temperature, CO2 and the stability of the microscopic stage are extremely important when performing these types of experiments, image acquisition with the MetaMorph® System Version 5.0 enormously simplified the amount of work associated with image acquisition and generating quality data. The fact that we have total control of the digital camera, exposure times and shutter wheels, makes it possible to create movies of living material with GFP-tagged proteins or other fluorescent dyes without damaging the sample.
Following fixation of the cells and immunofluorescence processing, images are acquired and a variety of measurements can be performed. I find this system very useful to study the number and size of cellular components and their distribution within the cell. For example, I monitor the fluorescence intensity of specific subregions in order to access the concentration of the labeled epitope within this region. Lately I have been using this system to analyze old confocal microscopic images that I’d previously acquired. After creating 3D reconstructions, I perform the same measurements as described above.
In conclusion, although expensive, the variety and versatility of applications and excellent control of the microscope and digital camera that the MetaMorph® System has to offer make its use ideal in any cell biology laboratory.