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zeiss_axiovert_200m [2016/04/06 00:25] bioimaging created |
zeiss_axiovert_200m [2016/06/08 17:36] bioimaging [Objectives] |
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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=axiovert_200m_name.jpg"> |
<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.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki.jpg" 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.jpg">Room P2-A-22</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=telephone_icon.gif" 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>Software</b>: Metamorph 7.7.9.0<br> |
- | <b>Year</b>: 2015 <br> | + | <b>Year</b>: 2003 <br> |
- | <b>SN</b>: 1026052056<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> |
+ | <br> | ||
+ | → <a href="/facility/bioimaging/doku.php?id=zeiss_axiovert_200m_repair"><img src="/facility/bioimaging/lib/exe/fetch.php?media=repair-icon.gif" width=16> Zeiss Axiovert 200M Repair History</a> | ||
</p> | </p> | ||
</td> | </td> | ||
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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 (Roper Scientific 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 our [[http://micro.magnet.fsu.edu/primer/digitalimaging/deconvolution/deconvolutionhome.html|deconvolution]] server [[brainstorm|Brainstorm]] 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_wf|Zeiss Cell Observer WF]] with Definite Focus. If don't want to use deconvolution, check out one of the confocal systems such as the [[zeiss_lsm_710|Zeiss LSM 710]], [[zeiss_lsm_880|Zeiss LSM 880]] or the [[3i_marianas_sdc|3i Marianas SDC]] spinning disk. 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. | ||
+ | |||
+ | ===== 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]]**. | ||
+ | - Users must indicate which lasers they are going to use in the booking Description field | ||
+ | - Users must indicate if they will use the incubator in the booking Description field (write down 37C INCUBATOR) | ||
+ | - Users must write their internal extension phone number in the booking Description field | ||
+ | |||
+ | ===== 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 ^ Type ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ (color camera) 10 μm = ^ | ||
+ | | 10x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420340-9901-000|EC Plan-Neofluar]] | Dry | 0.30 | 5.2 | 1.00 | 10 pixels | 23.3 pixels | | ||
+ | | 20x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|EC Plan-Neofluar]] | Dry | 0.50 | 2 | 0.48 | 21 pixels | 46.7 pixels | | ||
+ | | 40x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440350-9903-000|EC Plan-NeoFluar]] | Dry | 0.75 | 0.71 | 0.24 | 41 pixels | 93.7 pixels | | ||
+ | | 63x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420782-9900-000|Plan-Apochromat]] | Oil | 1.40 | 0.19 | 0.15 | 65 pixels | - | | ||
+ | |||
+ | |||
+ | **Upon request:** | ||
+ | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ | ||
+ | | 1.25x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=422310-9900-000|EC Epiplan-Neofluar]] | Dry | 0.035 | 4 | 7.58 | 1.3 pixels | | ||
+ | | 5x | [[https://www.micro-shop.zeiss.com/?s=571974962233f6&l=en&p=uk&f=o&a=v&m=s&id=420330-9901-000|EC Plan-NeoFluar]] | Dry | 0.16 | 18.5 | 2.00 | 5 pixels | | ||
+ | | 20x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440341-9904-000|EC Plan-Neofluar Ph 2]] | Dry | 0.50 | 2 | 0.48 | 21 pixels | | ||
+ | | 40x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440451-9903-000|Plan-Neofluar Ph3]] | Oil | 1.30 | 0.20 | - | - | | ||
+ | | 63x | [[https://www.micro-shop.zeiss.com/?s=571858182d6d9d&l=en&p=uk&f=o&a=v&m=s&id=420781-9910-000|Plan-Apochromat Ph3]] | Oil | 1.4 | 0.19 | 0.15 | 65 pixels | | ||
+ | | 100x | [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440782-9902-000|Plan-Apochromat]] | Oil | 1.40 | 0.17 | 0.10 | 102 pixels | | ||
+ | |||
+ | *** pixel sizes for binning = 1** | ||
+ | |||
+ | ==== Stage ==== | ||
+ | For automated acquisition of 96 well plates use: | ||
+ | X = -9014 μm | ||
+ | Y = -8995 μm | ||
+ | |||
+ | ===== System Turn On Procedures ===== | ||
+ | * Turn the fluorescence light power source on. | ||
+ | {{halide_710.jpg?80|}} | ||
+ | * Turn on the Uniblitz shutter controller (on the back). | ||
+ | {{ax_uniblitz1.jpg?100|}}{{ax_uniblitz2.jpg?100|}} | ||
+ | * Turn the Prior microscope stage controller on. | ||
+ | {{ax_prior_on.jpg?100|}} | ||
+ | * Wait 10s then turn the microscope on. | ||
+ | {{ax_mic_on.jpg?100|}} | ||
+ | * Turn the camera on. | ||
+ | {{ax_cam_on.jpg?100|}} | ||
+ | * Turn the computer on. | ||
+ | * Login to the computer (each group has it's own area and login). | ||
+ | * Start the Metamorph software. | ||
+ | * If you need temperature control, turn on the control box | ||
+ | {{heating2.jpg?100|}} | ||
+ | * If you need CO2, open the valve and turn on CO2 supply on the control box | ||
+ | {{co2.jpg?100|}} | ||
+ | |||
+ | ===== 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 shutter 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]] |