|Olympus VivaView Incubator/Fluorescence Microscope|
Just as its name implies, the Olympus VivaView FL incubator/fluorescence microscope consists of an inverted fluorescent microscope that is fully contained in a moderate-sized CO2 incubator. Such a configuration allows for extremely stable time lapse imaging in an environment that is identical to the most common in vitro cell culture conditions. The system also features a motorized stage for monitoring multiple positions in a dish, as well as an automated tray with space for up to 8 dishes. Consequently, whole experiments and a full set of controls can be precisely imaged in parallel for as long as biologically feasible - the final image number/file size is generally the only practical consideration.
Such long-term stability does come at an optical expense. The VivaView is limited to a single 40x objective (40x/0.95 NA), which can be further adjusted (with no gain in resolution) to either 20x or 80x via additional lenses. The objective is, however, well-suited for DIC (differential interference contrast) imaging, and produces transmitted light images of excellent quality. Basic fluorescent capture is also possible through a 200W mercury arc lamp X-Cite eXacte illumination system and a set of FITC (green) and TRITC (red) filter sets.
Transmitted light can become birefringent when passed through polystyrene (plastic) cultures dishes/lids. As a result, effective DIC imaging in the VivaView requires that cells be plated in certain round 35 mm glass-bottom dishes, which are available for purchase from the facility. In addition, the plastic culture dish lids must be replaced with specialized glass covers that are maintained by the facility staff. (Note that these lids are not disposable, and must be kept with the instrument at all times - they are merely swapped with incoming samples' lids immediately prior to imaging.)
The VivaView is controlled via a PC workstation running an aquisition version of the MetaMorph software. The microscope control interface is straightforward and intuitive, but the software itself does hide a number of complex tools that may not be required by most users. Capture is performed with an ORCA-R2 Hamamatsu cooled CCD camera (1344 x 1024 pixels).
In all, the VivaView has the potential to streamline several common imaging approaches, including wound healing/scratch assays, timed drug delivery, migration studies, and observations of fluorescent protein dynamics.
Currently illumination is possible only through DIC transmitted light imaging, as well as FITC (green) and TRITC (red) filter sets for fluorescent imaging. Additional fluorescent filters can be purchased and added if there is appropriate demand.
|Magnification||Model Name||Immersion Type||NA||Working Distance|
|40x (w/ 0.5x, 2x)||UPLSAPO 40x||dry||0.95||0.18 mm|
As mentioned above, the VivaView requires glass-bottom dishes for properly visualizing DIC modalities (see image, top left). The facility keeps several types of these dishes in stock; please inquire with a staff member if there is a need to purchase some prior to beginning experiments. These glass-bottom dishes come with a plastic lid, which must be exchanged with the reuseable glass lids (bottom left) when loading dishes in the incubator. These lids are sterilized on a regular basis as a service to users, but they must be left with the instrument at all times.
Note that there are actually two types of glass lids; one set includes a hinge on the back that clicks into place during sample loading. This lid allows for a motorized open/close control of a designated dish, and therefore enables pharmacological interventions via manual pipetting through the small opening on the internal door of the incubator (see image, right).
1. Place sample dishes in the incubator to begin the pre-warming process. The incubator itself should always be left on.
2. If using fluorescence, turn on the X-Cite eXacte lamp. (Otherwise, skip this step and ignore any warnings concerning the lamp when starting the software.)
3. Turn on the Hamamatsu camera controller by pushing and holding the power button for 2 seconds.
4. Log on to the PC username and password that was created during microscope training.
5. Double-click the VivaView FL software icon and begin loading sample dishes/lids into the imaging tray.
1. When finished with imaging, carefully remove samples from the incubator imaging tray. Use the software controls to rotate the tray, if necessary. Save all files, exit the VivaView FL program, and log off the computer.
2. Turn off the Hamamatsu camera controller by briefly pushing the power button.
3. If fluorescence was used, turn off the X-Cite eXacte lamp.
4. Leave the metal/glass dish covers on a paper towel above the incubator. A staff member will sterilize these non-disposable lids at the earliest convenience.
5. Do NOT power off the incubator.