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zeiss_axiovert_200m [2016/05/30 17:43] bioimaging |
zeiss_axiovert_200m [2024/02/12 22:50] bioimaging |
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- | <img src="/facility/bioimaging/lib/exe/fetch.php?media=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_axiovert_200m_wiki.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki.jpg" width=300></a> | + | <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_2022.png"><img src="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_wiki_2022.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_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>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=zeiss_axiovert_200m_map_new_p2a24.png">Room P2-A-24</a> (<img src="/facility/bioimaging/lib/exe/fetch.php?media=phone_neg.png" width=15> 47219) <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">ZEISS Microscopy</a><br> |
<b>Model</b>: Axiovert 200M<br> | <b>Model</b>: Axiovert 200M<br> | ||
- | <b>Software</b>: Metamorph 7.7.9.0<br> | + | <b>Nickname</b>: "Axiovert"<br> |
+ | <b>Software</b>: Metamorph 7.8.0.0<br> | ||
<b>Year</b>: 2003 <br> | <b>Year</b>: 2003 <br> | ||
<b>SN</b>: ZCA522000494<br> | <b>SN</b>: ZCA522000494<br> | ||
+ |   <br> | ||
+ | Data will be deleted after: <b>1 month</b> | ||
  <br> |   <br> | ||
  <br> |   <br> | ||
  <br> |   <br> | ||
- | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/new/3"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Axiovert 200M Booking</a> | ||
- | <br> | ||
→ <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> | → <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_axiovert_200m_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png"> Zeiss Axiovert 200M 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> | + | </p> |
- | → <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> | + | |
</td> | </td> | ||
</tr> | </tr> | ||
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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 (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. | + | {{ :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 ([[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics CoolSNAP HQ CCD]]) allows you to capture weak fluorescent signals and minimize photobleaching/photodamage in light sensitive samples. It is also equipped with a color camera ([[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]]) for acquisition of histological stains with tile and stitching imaging. |
- | ===== Booking Rules ===== | + | * **Microscope**: Zeiss Axiovert 200 |
- | The Axiovert 200M booking is divided in three daily slots: | + | * **Monochrome Camera**: [[https://labs.pbrc.edu/cellbiology/documents/CoolSNAPHQ.pdf|Photometrics Coolsnap HQ CCD]] |
- | | 9:00 to 13:00 | 13:00 to 17:00 | 17:00 to 21:00 | | + | * **Color Camera**: [[https://www.leica-microsystems.com/products/microscope-cameras/p/leica-dfc450-c/|Leica DFC450]] |
- | - A **user** can book at maximum **2 sessions per week** | + | {{: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. |
- | - 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 ===== | ===== System components ===== | ||
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==== Filtersets ==== | ==== Filtersets ==== | ||
- | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ 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 | | + | | 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 | 395 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 | | + | | 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 | 510 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 | | + | | 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 | 560 nm | 575-640 nm | |
- | | 4 | Far Red | [[http://www.semrock.com/SetDetails.aspx?id=2983|Cy5-4040C-000]] | 608-648 nm | 672-712 nm | | + | | 4 | Far Red | [[http://www.semrock.com/SetDetails.aspx?id=2983|Cy5-4040C-000]] | 608-648 nm | 660 nm | 672-712 nm | |
- | | 5 | A | none | none | none | | + | | 5 | A | none |||| |
**Upon request:** | **Upon request:** | ||
- | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Emission ^ | + | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ 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 | | + | | 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 | 510 nm | > 515 nm | |
==== Objectives ==== | ==== Objectives ==== | ||
- | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ (color camera) 10 μm = ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ (color) 10 μm = ^ Reference ^ |
- | | 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 | | + | | 10x | EC Plan-Neofluar | Air | 0.30 | 5.20 | 1.00 | 10 pixels | 23.3 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]] | |
- | | 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 | 46.7 pixels | | + | | 20x | EC Plan-Neofluar | Air | 0.50 | 2.00 | 0.48 | 21 pixels | 46.7 pixels | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440340-9904-000/Objective-EC-Plan-Neofluar-20x-0.5|440340-9904-000]] | |
- | | 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 | 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]] | |
- | | 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 | 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]] | |
- | | 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 | - | | + | | 100x | α Plan-Apochromat DIC | Oil | 1.46 | 0.11 | 0.09 | 105 pixels | - | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420792-9800-000/Objective-alpha-Plan-Apochromat-100x-1.46-Oil-DIC-M27|420792-9800-000]] | |
- | | 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 | - | | + | |
**Upon request:** | **Upon request:** | ||
- | ^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ Reference ^ |
- | | 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 | | + | | 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 | [[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 | | + | | 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]] | |
- | | 25x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420852-9972-000|LCI Plan-NeoFluar]] | Oil/Glyc/W | 0.8 | 0.21 | 0.40 | 25 pixels | | + | | 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]] | |
- | | 40x | [[https://www.micro-shop.zeiss.com/?s=129353259d52f12&l=en&p=uk&f=o&a=v&m=s&id=420660-9970-000|Plan-Apochromat]] | Dry | 0.95 | 0.25 | 0.25 | 40 pixels | | + | | 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]] | |
- | | 63x (Ph 3) | [[https://www.micro-shop.zeiss.com/?s=571858182d6d9d&l=en&p=uk&f=o&a=v&m=s&id=420781-9910-000|Plan-Apochromat]] | Oil | 1.4 | 0.19 | 0.15 | 65 pixels | | + | |
*** 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 (if used) |
{{halide_710.jpg?80|}} | {{halide_710.jpg?80|}} | ||
- | * Turn on the Uniblitz shutter controller (on the back). | + | * Turn on the Prior stage controller |
- | {{ax_uniblitz1.jpg?100|}}{{ax_uniblitz2.jpg?100|}} | + | |
- | * Turn the Prior microscope stage controller on. | + | |
{{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 (each group has it's own area and login). | + | * Log in to Windows (Bioimaging User) |
- | * Start the Metamorph software. | + | * Start the Metamorph software |
- | * If you need temperature control, turn on the control box | + | * If the software doesn't detect the camera, power cycle the camera and try again |
- | {{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 ===== | ===== 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 (if used) |
- | * 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 (if used) | ||
---- | ---- | ||
- | [[resources|Back to the Resources page]] | + | [[resources|Back to the Equipment page]] |