Zeiss LSM-900 confocal Airyscan microscope
Desmarais Building, Room 2234
Advanced Microscope Tier 2 usage price
Instrument awarded to Dr. Jean-Philippe Gratton by the Canadian Foundation for Innovation (CFI # 34992) in 2015
Applications
- Inverted microscope
- Airyscan super-resolution with joint deconvolution
- Point scanning confocal
- Brightfield
- Phase contrast
- Interference Contrast (DIC)
- Fluorescence
- Targeted photo-modulation FRAP
- Incubation
- Timelapse imaging
- Deconvolution
- Extended Depth of Focus
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Objectives
5x/0.15 Ph1 Air WD 5.30 N-AchroPlan 420931-9911
20x/0.80 Air WD 0.61 Plan ApoChromat 420650-9901
63x/1.40 Oil WD 0.19 Plan ApoChromat 420782-9900
| title | Complete lens specifications |
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Position
Nom
Marque
Nom complet
Identifiant
Grossissement
Ouverture numérique
Immersion
Type
Distance de travail (mm)
Transmittance
(% [nm])
Technique
Épaisseur du couvre-objet (mm)
2
20x/0.80
Air
Zeiss
20x/0.8 DIC II
Plan-Apochromat
W0.8x1/36"
20x
0.8
Air
Plan Apochromat
0.55
BF, DIC, Fluo
0.17
4
63x/1.40
Huile
Zeiss
63x/1.4 DIC III
Plan-Apochromat
M27
63x
1.4
| Note |
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| Huile |
Plan Apochromat
0.19
BF, DIC, Fluo
0.17
BF: Bright-field
DIC: Interference contrast
- Filter cubes
| title | Complete filter specifications |
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Position
Nom
Marque
Identifiant
Filtre d'excitation
Miroir dichroïque
Filtre d'émission
Commentaire
1
DAPI
Zeiss
365/50
[340-390]
395LP
445/50
[420-470]
2
GFP
Zeiss
470/40
[450-490]
495LP
525/50
[500-550]
FT 495
BP 525/50
- 2 GaASP PMT
| id | User Guide |
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| title | User Guide |
| expanded | true |
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| title | Startup |
Turn on the System (#2) and Components (#3) switches in the rack on the left of the microscope
Turn on the laser key (#4) in the rack on the left of the microscope
| Note |
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| When using for the first time, it is necessary to import the microscope-specific parameters BEFORE starting the software. See the First Use section below. |
Start the Zen software
| title | First Use |
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| Note |
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Please note, this procedure will delete all your experiment protocols and restore the software to its original settings. |
- If open, close the Zen software and wait for it to close completely (up to 30 seconds)
- On the Desktop open the Documentation folder
- Double-click Settings for LSM800
- A script will run and a black window will appear briefly
- You can then reopen the Zen software
| title | Loading samples |
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This procedure puts the microscope in a safe configuration and performs a focus calibration. At the end of this procedure the microscope will be ready for acquisition.
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On the microscope touch screen:
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| title | First focus |
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| Warning | ||
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Make sure to calibrate the focus before performing the first focus. |
On the microscope touch screen:
- Press Home>Microscope>Turret>Objectives
- Press 5x to select the 5x lens
Info The 5x objective is the safest because it has the longest working distance (TBD mm). The sample will appear perfectly sharp long before the lens approaches it. It is recommended to always first focus with the safest lens. The objectives are para-focal, focusing with the safest objective will then allow you to easily find your sample with another objective.
- Press Home>Load Position to lower the stage to its lowest position
- Press Set Work Position to store this position
- If necessary, move the focus slightly up to remove the “Lower Z limit reached” message displayed on the touchscreen
- Place the test slide on the microscope stage with the coverslip toward the objective
Note title Important Always use the test slide to perform the first focus.
- If necessary, move the stage so that the sample is centered on the objective
On the computer:
| Note |
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Once calibrated, the focus can be found at Z = 1.6 mm). The Z value can be found on the microscope touch screen Home>Z-Position |
| title | Seconday focus |
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| Warning | ||
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First focus with the safest lens before selecting another lens and continuing with secondary focus. |
| title | Focusing with air objectives |
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After performing the first focus, on the microscope touch screen:
| Info |
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| The 40x objective is the best Air objective because it has the greatest number of optical corrections (Plan Apochromat) and the largest numerical aperture (0.95). It offers a lateral resolution of 420nm at a wavelength of 550nm. |
| Note |
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There are two (2) 40x objectives, make sure you select the Air one |
| title | Focusing with oil lenses |
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After performing the first focus, on the microscope touch screen:
Press 63x Oil, 40x Oil to select the desired lens. The microscope will automatically lower the stage so that the sample is accessible.
| Info |
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The 40x objective is the best oil objective because it has the most optical corrections (Plan Apochromat) and the largest numerical aperture (1.4). It offers a lateral resolution of 240nm at a wavelength of 550nm. |
| Note |
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There are two (2) 40x objectives, make sure you select the OIL one |
In Zen software:
- In the Locate tab, select BF or the desired fluorescence (DAPI, GFP, mPlum) to activate the configuration
- Adjust the focus with the precision dial while looking through the eyepieces until the image is perfectly sharp
- In the Locate tab, select Off to turn the illumination off
- Your sample is ready for acquisition!
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| id | Light Path |
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| title | Light Path |
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| id | Log |
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| title | Log |
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| id | Technical Datasheet |
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| title | Technical Datasheet |
Stand
Zeiss AxioObserver LSM900 Serial: 2633000272
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- Slide combo
- 6-well plate
- 35 mm dish
- Multi-well plat
Filters
6-Position motorized Filter Turret 424947
- GFP Zeiss Filter Set 38 cube 424931 000000-1031-346 BP 470/40 450-490 FT 495 BP525/50 500-550
- DAPI Zeiss Filter Set 49 cube 424931 489064-0000-000 G365 FT 395 BP 445/50 335-383 395 420-470
- mPlum Zeiss Filter Set 64 HE cube 424931 489064-0000-000 BP587/25 FT605 BP647/70 574-599 605 612-682
- DIC Analyzer Zeiss 424937-9901
- Empty
- Empty
Scanner
LSM900 Scan head
- 8x Motorized Filter wheel 1
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- 8x Motorized Filter wheel 2
- Empty
- LBF 640 0000000-2090-299 410-617 93% 630-644 0.001%
- LP 575 0000000-2090-294 576-750 93% 410-565 0.001%
- LP 655 0000000-2090-295 656-750 93% 395-643 0.001%
- SP 620 000000-2090-298 410-617 93% 630-750 0.001%
- Empty
- Empty
- Empty
Main Beam splitter
MBS 405+488+561+640 T10/R90
400102-9600-000
Transmission
- 420-471 93%
- 502-546 93%
- 575-618 93%
- 656-800 93%
- 631-642 10%
Reflection
- 399-410 99%
- 481-492 99%
- 558-564 99%
- 631-642 90%
Detector
- 2 GaASp PMT
- 1 Airyscan 2 63x Serial 2657000116
- 1 transmitted ESID Motorized Left TL ESID LED, Right ESID Detector
Workstation
- HP Z2 G9 Serial CZC4027PF7 829W6EC#ABB
I155440-CIB
- Motherboard HP 895C
- BIOS version 2024-09-03
- 1 x Intel Core i7-12700K 3.6 GHz
- RAM 128GB DDR5 4 x 32GB no ECC
- OS 500 GB NVMe 6 GB/s
- 7 TB HD Data Storage 213 MB/s
- VNvidia RTX A4000 16 GB GDDR6
- Monitor HP Z32x
- Software Zen 3.10
Incubation
Consumables
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| id | FAQ |
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| title | Troubleshooting & FAQ |
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- Open the light path file
- Starting from the light source and moving towards the detector, verify that there is indeed light after each component of the microscope
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| id | Demo Image |
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| title | Zeiss LSM-800 |
| direction | horizontal |
| id | Demo Image |
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| title | Demo Image |
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