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--- zeiss_lsm_880 [2016/04/04 16:43]
bioimaging
+++ zeiss_lsm_880 [2017/04/13 18:31]
bioimaging [Microscope overview]
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 &nbsp <br>
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-&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/resource_admin.php?resourcepage=0&rid=128"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 880 Booking</a>
+&nbsp <br>
+&rarr; <a href="https://imm.medicina.ulisboa.pt/intranet/booking/128"><img src="/facility/bioimaging/lib/exe/fetch.php?media=date.png">  Zeiss LSM 880 Booking</a>
  <br>
 &rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Zeiss LSM 880 Quality Control</a>
  <br>
 &rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Zeiss LSM 880 Usage Statistics</a>
-&nbsp </br>
-&rarr; <a href="/facility/bioimaging/doku.php?id=zeiss_lsm_880_repair"><img src="/facility/bioimaging/lib/exe/fetch.php?media=repair-icon.gif" width=16> Zeiss LSM 880 Repair History</a>
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 ===== Microscope overview =====
-The Zeiss LSM 7 Live //confocal line-scanning microscope// is a fast imaging system able to analyze high-speed processes with a time resolution of just a few microseconds. It is an upright microscope equipped with water dipping objectives specially suited for live samples in aqueous media. The scanning unit uses a laser beam with a rectangular cross-section to [[http://www.nature.com/nmeth/journal/v3/n11/thumbs/nmeth971-F2.jpg|illuminate a whole line in the sample]], instead of a single point. This makes image acquisition by its array CCD detectors faster than conventional PMT light detection but it also lowers the Z-resolution due to the usage of slits instead of conventional pinholes. Note that the system is equipped with two lasers (488 and 561 nm) but dual color imaging is not advised since there are no bandpass filters in the system (channel crosstalk is likely to occur). A UV laser controlled by galvanometric mirrors is available for precise ablation in user-defined regions of interest. The spatial resolution of this system is not as high as a //point-scanning confocal system// such as the [[zeiss_lsm_880|Zeiss LSM 880]] or [[zeiss_lsm_710|Zeiss LSM 710]] but it is much faster than these two systems, achieving scanning speeds which may be even higher than the [[3i_marianas_sdc|3i Marianas SDC]] spinning disk if the acquisition is performed only in a few lines. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].
+{{  :airyscan.jpg?0x380}} The Zeiss LSM 880 with Airyscan is a //confocal point-scanning microscope// able to generate high-resolution three-dimensional images of thick specimens with high sensitivity and low photodamage. It is an inverted microscope specially suitable for live cell imaging and photobleaching experiments, equipped with a large size incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. Its [[http://www.zeiss.com/content/dam/Microscopy/Products/laser-scan/LSM%20880/zeiss-lsm880-airyscan-principle-672.jpg|Airyscan]] 32 channel area detector is able to collect all light from an Airy pattern simultaneously, allowing for increased resolution (140 nm laterally and 400 nm axially, at 488 nm). Its scanning unit further includes two PMT detectors (one of them cooled) as well as a GaAsP detector for increased sensitivity (45% QE compared to ~25% QE for conventional PMT).  It is equipped with lasers from violet to far red (405, 458, 488, 514, 561, 594 and 633 nm excitation wavelengths). With this system you can perform optical sectioning of fluorescent samples which are too thick for a widefield system such as the [[zeiss_cell_observer|Zeiss Cell Observer]] or the [[zeiss_axiovert_200m|Zeiss Axiovert 200M]]. Image resolution with Airyscan is higher than the [[zeiss_lsm_710|Zeiss LSM 710]] and detection sensitivity is also higher than faster confocal systems such as the spinning disk [[3i_marianas_sdc|3i Marianas SDC]] or the //line scanning// [[zeiss_lsm_7_live|Zeiss LSM 7 Live]]. If your personal computer cannot handle all the data you collected, check out the [[big_guy|Big Guy]] or [[colossus|Colossus]].
-{{info.gif?|}} **If you are going to use the 561 nm laser please write '561' in the Event Field name**
+{{info.gif?|}} Click on the image on the left to see the system beam path in higher detail
+===== Booking Rules =====
+The Zeiss LSM 710 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)
 ===== System components =====
 ==== LASERs ====
 ^  Laser Unit  ^  Wavelength  ^  Maximum Power  ^  Current Status  ^
-|  Sapphire 488-100  |  488 nm  |  100 mW  |  ok  |
+|  Diode 405-30  |  405 nm  |  30 mW  |  ok  |
-|  DPSS 561-40  |  561 nm  |  40 mW  |  ok  |
+|  Argon  |  458, 488 and 514 nm  |  25 mW  |  ok  |
+|  DPSS 561-20  |  561 nm  |  20 mW  |  ok  |
+|  HeNe594  |  594 nm  |  2 mW  |  ok  |
+|  HeNe633  |  633 nm  |  5 mW  |  ok  |
 ==== Objectives ====
-^  Magnification  ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
+^  Magnification   ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
-|  10x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440039-0000-000|W Achroplan Ph1]]  |  Water dipping  |  0.30  |  3.1  |
+|  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  |
-|  10x  |  [[https://www.micro-shop.zeiss.com/?s=2065696c9e13e&l=en&p=uk&f=o&a=v&m=s&id=440330-0000-000|Plan-Neofluar]]  |  Dry  |  0.30  |  5.5  |
+|  20x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=420650-9901-000|Plan-Apochromat]]  |  Dry  |  0.80  |  0.55  |
-|  20x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=421452-9900-000|W Plan-Apochromat]]  |  Water  |  1.00  |  1.8  |
+|  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  |  0.80  |  0.21  |
-|  40x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440090-9901-000|W Achroplan]]  |  Water dipping  |  0.80  |  3.6  |
+|  40x  |  [[https://www.micro-shop.zeiss.com/?s=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=421767-9970-000|C-Apochromat]]  |  Water  |  1.20  |  0.28  |
-|  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=3407786617d462&l=en&p=uk&f=o&a=v&m=s&id=440762-9904-000|Plan-Apochromat]]  |  Oil  |  1.40  |  0.19  |
+|  100x  |  [[https://www.micro-shop.zeiss.com/?s=7138707e1a1de&l=en&p=uk&f=o&a=v&m=s&id=420792-9800-000|alpha Plan-Apochromat]]  |  Oil  |  1.46  |  0.11  |
+**Upon request:**
+^  Magnification   ^  Model  ^  Type  ^  NA  ^  WD (mm)  ^
+|  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  |
 ==== Filtersets (Ocular) ====
 ^  Position  ^  Filterset  ^  Reference  ^  Excitation  ^  Emission  ^
-|  2  |  Green  |  [[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  |
+|  1  |  Green  |  [[https://www.micro-shop.zeiss.com/?s=7139069fdc79c&l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&o=|FS38HE]]  |  450-490 nm  |  500-550 nm  |
-|  3  |  Red  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488015-0000-000&o=|FS15]]  |  540-552 nm  |  > 590 nm  |
+|  2  |  Red  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=000000-1114-101&o=|FS43]]  |  533-558 nm  |  570-640 nm  |
-|  4  |  Blue (for ablation*)  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]]  |  none  |  420-470 nm  |
+|  3  |  Blue  |  [[https://www.micro-shop.zeiss.com/?s=163176628fcf844&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]]  |  G 365 nm  |  420-470 nm  |
-* Excitation filter has been removed
-==== Filter and Dichroic sets ====
+===== Microscope Turn On Procedure =====
-|  {{zeiss5livefilters0.jpg?100}}  |  {{zeiss5livefilters1.jpg?100}}  |
+  * Turn on the ''MAIN SWITCH''
-|  Main configuration  |  Emission filters  |
+  * Turn on the two switches: ''SYSTEMS/PC'' and ''COMPONENTS''
+{{zeiss_lsm_880_1.jpg?100}}
+  * Turn the computer on
+  * Login with your unit account
+  * Wait for network icon to change to ''connected'' state
+  * Start the **ZEN** software
-===== Microscope Turn On Procedures =====
+===== Microscope Turn Off Procedure =====
+If there's another user for this microscope in the next hour:
-==== Normal Operation ====
+  * Close the software, leave the lasers on and log off the computer.
+  * Clean up any objective where you used immersion oil.
-  * The system must be turned on **30 min** before usage.
-    * To start it turn on the two main switches: ''SYSTEM/PC'' and ''COMPONENTS''
-{{7live_switches.jpg?100|}}
-  * Login to the computer, wait for the network icon to change to ''connected'' state
-  * Start the **ZEN 2009** software.
-**After the 30 min:**
-  * Turn on the metal halide lamp (white box to the left of the microscope).
-{{7live_excite.jpg?100|}}
-  * You may now start using the system.
-==== Live Imaging / Time Lapse Operation ====
-  * The system must be turned on **2 hours** before usage (in order to properly warm up scanning mirrors).
-    * To start it turn on the two main switches: **SYSTEM/PC** and **COMPONENTS**
-  * Login to the computer, wait for the network icon to change to ''connected'' state
-  * Start the **ZEN 2009** software.
-**After the 2 hours:**
-  * Turn on the metal halide lamp (white box to the left of the microscope).
-  * You may now start using the system.
-===== Microscope Turn Off procedures =====
-If there's another user for this microscope in the next two hours:
-  * Exit the **ZEN 2009** software, leave the lasers on and log off the computer.
-  * Clean up immersion objectives.
   * **Make sure that there really is another user going to use the microscope.**
 Else:
-  * Turn off the lasers, wait 5 min for them to cooldown.
+  * Turn off the lasers on **ZEN**
-  * Exit the **ZEN 2009** software and shut down the computer.
+  * Clean up any objective were you used immersion oil.
-  * Clean up immersion objectives.
+  * Exit the software, shut down the computer.
-  * Turn off the metal halide lamp.
+  * Turn off the main switches ''SYSTEMS/PC'' and ''COMPONENTS''
-  * Turn off the microscope.
+  * Wait 5 min. for Ar laser cooldown
-  * Turn off the two main switches: **SYSTEM/PC** and **COMPONENTS**
+  * Turn off the ''MAIN SWITCH''.
-===== If you want to use the UV laser ablation system: =====
-  * Please contact the [[about|Bioimaging Facility]] (Ext: 47316). The ablation system requires specific training even for users that are already trained on the LSM 7 Live itself.
 ----
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