This shows you the differences between two versions of the page.
Both sides previous revision Previous revision Next revision | Previous revision Next revision Both sides next revision | ||
zeiss_axiovert_200m [2016/04/06 00:41] bioimaging |
zeiss_axiovert_200m [2016/06/08 17:31] bioimaging [Objectives] |
||
---|---|---|---|
Line 15: | Line 15: | ||
  <br> |   <br> | ||
  <br> |   <br> | ||
- | → <a href="https://imm.medicina.ulisboa.pt/intranet/booking/month.php?user=AXIOVERT&rid=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/3"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png"> Zeiss Axiovert 200M Booking</a> |
<br> | <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> | ||
Line 30: | Line 30: | ||
===== 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 [[bioimaging:bigguy|Big Guy]]. If you have FISH or other fixed fluorescence slides try the [[bioimaging:leica_dm5000b|Leica DM5000B]] instead. | + | 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. |
- | + | ||
- | {{:bioimaging:info.gif?|}} **If you are not going to use fluorescence please write 'TL' in the Event Field name** | + | |
===== Booking Rules ===== | ===== Booking Rules ===== | ||
The Axiovert 200M booking is divided in three daily slots: | 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| | + | | 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** | - A **user** can book at maximum **2 sessions per week** | ||
Line 66: | Line 64: | ||
^ Magnification ^ Model ^ Type ^ NA ^ WD (mm) ^ pixel size* (μm) ^ 10 μm = ^ (color camera) 10 μm = ^ | ^ 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 | | | 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=440341-9904-000|EC Plan-Neofluar Ph 2]] | Dry | 0.50 | 2 | 0.48 | 21 pixels | 46.7 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 | | | 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 | | | 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 | - | | | 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 | [[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:** | ||
Line 76: | Line 73: | ||
| 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 | [[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 | | | 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 | | ||
- | | 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 | | + | | 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=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 | [[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 | |
- | | 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 | | + | | 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** | *** pixel sizes for binning = 1** | ||
Line 89: | Line 86: | ||
===== System Turn On Procedures ===== | ===== System Turn On Procedures ===== | ||
* Turn the fluorescence light power source on. | * Turn the fluorescence light power source on. | ||
- | {{:bioimaging:halide_710.jpg?80|}} | + | {{halide_710.jpg?80|}} |
* Turn on the Uniblitz shutter controller (on the back). | * Turn on the Uniblitz shutter controller (on the back). | ||
- | {{:bioimaging:ax_uniblitz1.jpg?100|}}{{:bioimaging:ax_uniblitz2.jpg?100|}} | + | {{ax_uniblitz1.jpg?100|}}{{ax_uniblitz2.jpg?100|}} |
* Turn the Prior microscope stage controller on. | * Turn the Prior microscope stage controller on. | ||
- | {{:bioimaging:ax_prior_on.jpg?100|}} | + | {{ax_prior_on.jpg?100|}} |
* Wait 10s then turn the microscope on. | * Wait 10s then turn the microscope on. | ||
- | {{:bioimaging:ax_mic_on.jpg?100|}} | + | {{ax_mic_on.jpg?100|}} |
* Turn the camera on. | * Turn the camera on. | ||
- | {{:bioimaging:ax_cam_on.jpg?100|}} | + | {{ax_cam_on.jpg?100|}} |
* Turn the computer on. | * Turn the computer on. | ||
* Login to the computer (each group has it's own area and login). | * Login to the computer (each group has it's own area and login). | ||
* Start the Metamorph software. | * Start the Metamorph software. | ||
* If you need temperature control, turn on the control box | * If you need temperature control, turn on the control box | ||
- | {{:bioimaging:heating2.jpg?100|}} | + | {{heating2.jpg?100|}} |
* If you need CO2, open the valve and turn on CO2 supply on the control box | * If you need CO2, open the valve and turn on CO2 supply on the control box | ||
- | {{:bioimaging:co2.jpg?100|}} | + | {{co2.jpg?100|}} |
===== Microscope Turn Off procedures ===== | ===== Microscope Turn Off procedures ===== |