zeiss_axiovert_200m
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zeiss_axiovert_200m [2016/04/06 00:25] bioimaging created |
zeiss_axiovert_200m [2020/01/14 17:42] bioimaging [System Turn On Procedures] |
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| - | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_name.jpg"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_name_new.png"> |
| <table style="border:0px solid white;"> | <table style="border:0px solid white;"> | ||
| <tr style="border:0px solid white;"> | <tr style="border:0px solid white;"> | ||
| <td style="border:0px solid white;"> | <td style="border:0px solid white;"> | ||
| - | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wf_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wf_wiki.jpg" width=300></a> | + | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_new.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_new.png" width=300></a> |
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_cell_observer_wf_map.jpg">Room P3-A-10</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" width=15> 47311) <br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_map_new.png">Room P2-A-22</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47217) <br> |
| <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br> | <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">Carl Zeiss MicroImaging</a><br> | ||
| - | <b>Model</b>: Cell Observer<br> | + | <b>Model</b>: Axiovert 200M<br> |
| - | <b>Software</b>: ZEN 2 (blue edition)<br> | + | <b>Nickname</b>: "Axiovert"<br> |
| - | <b>Year</b>: 2015 <br> | + | <b>Software</b>: Metamorph 7.7.9.0<br> |
| - | <b>SN</b>: 1026052056<br> | + | <b>Year</b>: 2003 <br> |
| + | <b>SN</b>: ZCA522000494<br> | ||
|   <br> |   <br> | ||
|   <br> |   <br> | ||
|   <br> |   <br> | ||
| - |   <br> | + | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/3"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Axiovert 200M Booking</a> |
| - | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/month.php?user=ZEISSCELL1&rid=125"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Cell Observer WF Booking</a> | + | |
| <br> | <br> | ||
| - | → <a href="/facility/bioimaging/doku.php?id=zeiss_cell_observer_wf_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Cell Observer WF Quality Control</a> | + | → <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert200m_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss Axiovert 200M Quality Control</a> |
| <br> | <br> | ||
| - | → <a href="/facility/bioimaging/doku.php?id=zeiss_cell_observer_wf_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss Cell Observer WF Usage Statistics</a> | + | → <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert_200m_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss Axiovert 200M Usage Statistics</a> |
| - | </p> | + | </p> |
| </td> | </td> | ||
| </tr> | </tr> | ||
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| + | ===== 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. | ||
| + | |||
| + | * **Microscope**: Zeiss Axiovert 200 | ||
| + | * **Monochrome Camera**: [[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics Coolsnap HQ CCD]] | ||
| + | * **Color Camera**: [[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]] | ||
| + | |||
| + | {{:warning.png?25|}} Data files older than **1 month** will be automatically deleted on this system, please copy your data to the iMM server using the desktop link. | ||
| + | |||
| + | ===== Booking Rules ===== | ||
| + | The Axiovert 200M booking is divided in three daily slots: | ||
| + | | 9:00 to 13:00 | 13:00 to 17:00 | 17:00 to 21:00 | | ||
| + | |||
| + | - A **user** can book at maximum **2 sessions per week** | ||
| + | - Each **session must fit inside one of the three daily slots** (i.e. a session booked from 11:00 to 14:00 counts as 2 sessions) | ||
| + | - The above rules do not apply for **weekends** and for work days **before 9:00** and **after 21:00** | ||
| + | - The above rules do not apply for sessions booked on the very same day or **24 hours ahead** | ||
| + | - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**. | ||
| + | |||
| + | ===== System components ===== | ||
| + | |||
| + | ==== Filtersets ==== | ||
| + | |||
| + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | ||
| + | | 1 | Blue | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488001-9901-000&o=|FS01]] | 359-371 nm | > 397 nm | | ||
| + | | 2 | Green BP | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488010-9901-000&o=|FS10]] | 450-490 nm | 515-565 nm | | ||
| + | | 3 | Red | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488020-9901-000&o=|FS20]] | 540-552 nm | 575-640 nm | | ||
| + | | 4 | Far Red | [[http://www.semrock.com/SetDetails.aspx?id=2983|Cy5-4040C-000]] | 608-648 nm | 672-712 nm | | ||
| + | | 5 | A | none | none | none | | ||
| + | |||
| + | **Upon request:** | ||
| + | |||
| + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | ||
| + | | 2 | Green LP | [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488009-9901-000&o=|FS09]] | 450-490 nm | > 515 nm | | ||
| + | |||
| + | ==== Objectives ==== | ||
| + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ (color) 10 μm = ^ Reference ^ | ||
| + | | 10x | EC Plan-Neofluar | Air | 0.30 | 5.2 | 1.00 | 10 pixels | 23.3 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9901-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] | | ||
| + | | 20x | EC Plan-Neofluar | Air | 0.50 | 2 | 0.48 | 21 pixels | 46.7 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420350-9900-000/Objective-EC-Plan-Neofluar-20x-0.50-M27|420350-9900-000]] | | ||
| + | | 40x | EC Plan-NeoFluar | Air | 0.75 | 0.71 | 0.24 | 41 pixels | 93.7 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440350-9903-000/Objective-EC-Plan-Neofluar-40x-0.75|440350-9903-000]] | | ||
| + | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | 0.15 | 65 pixels | - | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-000/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-000]] | | ||
| + | |||
| + | |||
| + | **Upon request:** | ||
| + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ Reference ^ | ||
| + | | 1.25x | EC Epiplan-Neofluar | Air | 0.035 | 4 | 7.58 | 1.3 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/422310-9900-000/Objective-EC-Epiplan-Neofluar-1.25x-0.03-M27|422310-9900-000]] | | ||
| + | | 5x | EC Plan-NeoFluar | Air | 0.16 | 18.5 | 2.00 | 5 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420330-9901-000/Objective-EC-Plan-Neofluar-5x-0.16-M27|420330-9901-000]] | | ||
| + | | 20x | EC Plan-Neofluar Ph 2 | Air | 0.50 | 2 | 0.48 | 21 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440341-9904-000/Objective-EC-Plan-Neofluar-20x-0.5-Ph2|440341-9904-000]] | | ||
| + | | 40x | Plan-Neofluar Ph3 | Oil | 1.30 | 0.20 | 0.24 | 41 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440451-9903-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-Ph3|440451-9903-000]] | | ||
| + | | 63x | Plan-Apochromat Ph3 | Oil | 1.4 | 0.19 | 0.15 | 65 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420781-9910-000/Objective-Plan-Apochromat-63-1.4-Oil-Ph3-M27|420781-9910-000]] | | ||
| + | |||
| + | *** 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 ==== | ||
| + | For automated acquisition of 96 well plates use: | ||
| + | X = -9014 μm | ||
| + | Y = -8995 μm | ||
| + | |||
| + | ===== System Turn On Procedures ===== | ||
| + | * Turn on the fluorescent light power source | ||
| + | {{halide_710.jpg?80|}} | ||
| + | * Turn on the Prior stage controller | ||
| + | {{ax_prior_on.jpg?100|}} | ||
| + | * Turn on the microscope | ||
| + | {{ax_mic_on.jpg?100|}} | ||
| + | * Turn on the CCD camera | ||
| + | {{ax_cam_on.jpg?100|}} | ||
| + | * Turn on the computer | ||
| + | * Log in to Windows (Bioimaging User) | ||
| + | * Start the Metamorph software | ||
| + | * If the software doesn't detect the camera, power cycle the camera and try again | ||
| + | ===== Microscope Turn Off procedures ===== | ||
| + | * Turn off the computer. | ||
| + | * Turn off the camera. | ||
| + | * If you used any __immersion oil in the objectives, clean them up__. | ||
| + | * Turn off the microscope stage controller. | ||
| + | * Turn off the microscope. | ||
| + | * If there's __another user__ for this microscope in the __next hour__ leave the mercury __lamp power source on, else turn it off__. | ||
| + | * 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. | ||
| + | |||
| + | ---- | ||
| + | [[resources|Back to the Resources page]] | ||
zeiss_axiovert_200m.txt · Last modified: 2024/02/12 22:50 by bioimaging
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