The Zeiss 710 confocal scan head is coupled to the AxioObserver Z1 inverted microscope. This inverted microscope body provides versatility for live and fixed, slide, well-plate, and dish imaging applications. The Z1 is a fully motorized microscope, allowing touchscreen or software control of objectives; fluorescent cubes; transmitted and epi-fluorescent light features. Advanced features include tile scanning, positional memory, timelapse acquisition, live tissue temperature and CO2 control, spectral imaging mode, and AOTF-based rapid line switching acquisition.
Detailed Features:
Fluorescent excitation – Laser lines available on on this microscope include 405nm (25mW diode), 440nm (25 mW diode), 458nm, 488nm, 514nm (35mW Argon laser), 561nm (25mW diode), 594nm (2mWHeNe laser), and 633nm (5mW HeNe laser).
Widefield Excitation – The AxioObserver Z1 is equipped with four fluorescent filter cubes, Zeiss sets 49 (blue), 38 (green), 49 (red), and 76 HE (far red) which allows conventional fluorescent imaging of multi-channel emitting samples. Full details about these Zeiss filter sets, as well as a list of fluorophores visible with each cube, are available here.
Optics – The microscope is outfitted with a range of dry and immersion objectives, all with high numerical aperture and specifically designed for confocal imaging. These include EC Plan-Neofluar 10X (0.45NA), Plan-Apochromat 20X (0.80NA), and Plan-Apochromat 40X (0.95NA) dry objectives, Plan-Apochromat 40X (1.3NA) and Plan-Apochromat 63X (1.4NA) oil-immersion objectives, and a Plan-Apochromat 63X (1.3NA) water-immersion objective.
Live imaging – A full live-cell incubation system is also available. This includes a temperable stage capable of heating slides or dishes, modules to control temperature, carbon dioxide and humidity (Temp Module S1, CO2 module S1, O2 module S1, and heating device humidity S1),an enclosure to maintain the environment (Incubator PM S1) and a temperable objective ring.
Software – Image acquisition and analysis for the LSM 710 is controlled using Zen software. In addition to the basic image acquisition and analysis modules, we have four add-on modules for advanced imaging functions. Modules for FRET, FRAP, and Physiology allow the acquisition and analysis of these dynamic microscopic imaging techniques. A module called Multi-time allows for advanced control of multi-dimensional image capture sequences.
Funding Support
The purchase of this laser scanning confocal microscope and live cell imaging accessories was made possible by two NSF Major Research Instrumentation awards, in 2007 and 2010. These awards were the result of interdisciplinary efforts led by the PI, Dr. Anita McCauley, and faculty from Biology (Drs. Conner, Fahrbach, Johnson, and Muday), Chemistry (Dr. King), Physics (Drs. Bonin, Guthold and Williams), and faculty from WFUHS (Drs. Poole and Scarpinato). The university has renovated Winston Hall Room 009 in order to create a dedicated room for confocal microscopy and related sample preparation and teaching.
Applications
The LSM 710 is being used for a wide range of research and educational programs. Research uses include imaging of fluorescent protein expression in plant roots and stems, fluorescent protein and immunohistochemical label in Drosophila nervous system, morphological analysis of neuronal structure in both culture and brain sections, and FRAP and colocalization studies of DNA mismatch repair proteins in living cells. The microscope has also been utilized in undergraduate laboratory courses, Optics — taught by grant co-PI Richard Williams, Biology 214: Cell Biology, and Biology 369: The Cell Biological Basis of Disease, and a graduate course, Biology 775: Microscopy in the Biological Sciences.
Example Images
Earthworm epithelial sensory organs highlighted by anti-tubulin fluorescence
Honey Bee mushroom body labelled with anti-synapsin-I fluorescence