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zeiss_axiovert_200m [2020/01/01 22:08] bioimaging [System Turn On Procedures] |
zeiss_axiovert_200m [2020/01/20 23:39] bioimaging |
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===== Microscope overview ===== | ===== Microscope overview ===== | ||
- | The Zeiss Axiovert 200M is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging applications. It is equipped with a small stage incubator for temperature control and CO2 supply. Using the Metamorph software you can control the motorized filters, shutters and stage to simultaneously set up multi-color time-lapse imaging experiments in multiple stage positions. Its sensitive cooled CCD camera (Photometrics Coolsnap HQ CCD) allows you to capture weak fluorescent signals and minimize photobleaching/photodamage in light sensitive samples. You can also perform Z-stack acquisition and use [[https://huygens.i3s.up.pt/login.php|PPBI Huygens RM]] [[http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html|deconvolution]] server to significantly improve image resolution even in thick samples. If you are performing long time-lapse experiments in live samples (aqueous media) and need hardware focus control, check the [[zeiss_cell_observer|Zeiss Cell Observer]] with Definite Focus. If don't want to use deconvolution, check out one of the confocal systems such as the point scanners [[zeiss_lsm_710|Zeiss LSM 710]] and [[zeiss_lsm_880|Zeiss LSM 880]] or the spinning disks [[3i_marianas_sdc|3i Marianas SDC]] and [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead. | + | {{ :widefield_lightpath.png?0x300}}The Zeiss Axiovert 200M is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging applications. Using the Metamorph software you can control the motorized filters, shutters and stage to simultaneously set up multi-color time-lapse imaging experiments in multiple stage positions. Its sensitive cooled CCD camera (Photometrics Coolsnap HQ CCD) allows you to capture weak fluorescent signals and minimize photobleaching/photodamage in light sensitive samples. You can also perform Z-stack acquisition and use [[https://huygens.i3s.up.pt/login.php|PPBI Huygens RM]] [[http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html|deconvolution]] server to significantly improve image resolution even in thick samples. If you are performing long time-lapse experiments in live samples (aqueous media) and need hardware focus control, check the [[zeiss_cell_observer|Zeiss Cell Observer]] with Definite Focus or the [[nikon_eclipse_ti|Nikon Eclipse Ti]]. If don't want to use deconvolution but need optical sectioning, check out one of the confocal systems such as the point scanners [[zeiss_lsm_710|Zeiss LSM 710]] and [[zeiss_lsm_880|Zeiss LSM 880]] or the spinning disks [[3i_marianas_sdc|3i Marianas SDC]] and [[zeiss_cell_observer_sd|Zeiss Cell Observer SD]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]]. If you have FISH or other fixed fluorescence slides try the [[leica_dm5000b|Leica DM5000B]] instead. |
* **Microscope**: Zeiss Axiovert 200 | * **Microscope**: Zeiss Axiovert 200 | ||
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*** pixel sizes for binning = 1** | *** pixel sizes for binning = 1** | ||
+ | |||
+ | ==== Cameras ==== | ||
+ | ^ Model ^ Type ^ Frame Size ^ Pixel Size (µm) ^ | ||
+ | | [[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics Coolsnap HQ]] | CCD | 1392 x 1040 | 6.45 x 6.45 | | ||
+ | | [[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]] | color CCD | 2560 × 1920 | 3.4 x 3.4 | | ||
==== Stage ==== | ==== Stage ==== | ||
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===== System Turn On Procedures ===== | ===== System Turn On Procedures ===== | ||
- | * Turn the fluorescence light power source on. | + | * Turn on the fluorescent light power source |
{{halide_710.jpg?80|}} | {{halide_710.jpg?80|}} | ||
- | * Turn the Prior microscope stage controller on. | + | * Turn on the Prior stage controller |
{{ax_prior_on.jpg?100|}} | {{ax_prior_on.jpg?100|}} | ||
- | * Wait 10s then turn the microscope on. | + | * Turn on the microscope |
{{ax_mic_on.jpg?100|}} | {{ax_mic_on.jpg?100|}} | ||
- | * Turn the camera on. | + | * Turn on the CCD camera |
{{ax_cam_on.jpg?100|}} | {{ax_cam_on.jpg?100|}} | ||
- | * Turn the computer on. | + | * Turn on the computer |
- | * Login to the computer (Bioimaging user). | + | * Log in to Windows (Bioimaging User) |
- | * Start the Metamorph software. | + | * Start the Metamorph software |
+ | * If the software doesn't detect the camera, power cycle the camera and try again | ||
===== Microscope Turn Off procedures ===== | ===== Microscope Turn Off procedures ===== | ||
- | * Turn off the computer. | + | |
- | * Turn off the camera. | + | If there is another user for this microscope in the __next hour__: |
- | * If you used any __immersion oil in the objectives, clean them up__. | + | * Leave the fluorescent lamp on |
- | * Turn off the microscope stage controller. | + | * Clean up immersion objectives |
- | * Turn off the shutter controller. | + | Else: |
- | * Turn off the microscope. | + | * Clean up immersion objectives |
- | * If there's __another user__ for this microscope in the __next hour__ leave the mercury __lamp power source on, else turn it off__. | + | * Shutdown the computer |
- | * Be extra careful in weekends and at the end of the day to make sure that the mercury lamp is not going to be turned on for an excessive time. | + | * Turn off the camera |
+ | * Turn off the Prior stage controller | ||
+ | * Turn off the microscope | ||
+ | * Turn off the fluorescent lamp | ||
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[[resources|Back to the Resources page]] | [[resources|Back to the Resources page]] |