- Created by Nicolas Stifani, last modified on Aug 01, 2021
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Nikon Ti2-E fully motorized inverted microscope
Roger Gaudry R-619
Applications
Brightfield
Polarized light
- DIC
Fluorescence
- Live imaging
- Long timelapse imaging
Light sources
LED lamp for transmitted light
Lumencor SpectraX laser fluorescence
Filter name | Excitation Filter | Type | Working distance (mm) | Transmittance (% [nm]) |
---|---|---|---|---|
R | 640/30-25 | Plan Fluor | 2.1 | >75% [400-800] |
G | 470/24-25 | Excitation Filter | Dichroic mirror | Emission Filter |
B | 440/20-25 | 350/50x [325-375] | 400LP | 460/50m [435-485] |
NT | 510/25-25 | Plan Apo Lambda | 0.13 | |
V | 395/25-25 | |||
GY | 550\15-25 | Plan Apo Lambda | TBD | |
Storage GY | 575\25-25 |
Objectives
- 20x/0.5 Air Ph1
- 60x/1.4 Oil DIC WD 0.13
- 100x/1.45 Oil Ph3 WD 0.13
- 100x/1.45 Oil DIC WD 0.13
- Empty
- 20x\0.75 Air DIC
Position | Name | Brand | ID | Magnification | Numerical Aperture | Immersion | Type | Working distance (mm) | Transmittance (% [nm]) | Technique | Cover glass thickness (mm) |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | 20x/0.5 Air Ph1 | Nikon | 20x/0.5 Plan Fluor WD 2.1 Ph1? Air | 20x | 0.5 | Air | Plan Fluor | 2.1 | >75% [400-800] | BF, Pol, PhC, Fluo | 0.17 |
2 | 60x/1.4 Oil DIC | Nikon | 60x/1.4 Plan Apo Lambda DIC N2 | 60x | 1.4 | Oil | Plan Apo Lambda | 0.13 | >80% [400-750] | BF, Pol, DIC, Fluo | 0.17 |
3 | 100x/1.45 Oil Ph3 | Nikon | 100x/1.45 Plan Apo Lambda Ph3 | 100x | 1.45 | Oil | PPlan Apo Lambda | 0.13 | >75% [400-800] | BF, Pol, DIC, Fluo | 0.17 |
4 | 100x/1.45 Oil DIC | Nikon | 100x/1.45 Plan Apo Lambda DIC N2 | 100x | 1.45 | Oil | Plan Apo Lambda | 0.13 | BF, Pol, DIC, Fluo | ||
5 | Empty | ||||||||||
6 | 20x/0.75 Air DIC | Nikon | 20x/0.75 Plan Apo Lambda DIC N2 | 20x | 0.75 | Air | Plan Apo Lambda | TBD | BF, Pol, DIC, Fluo |
Filter cubes
DAPI/Hoechst/AMCA
FITC/EGFP
TRITC/Rhodamine
TexasRed
Position | Cube name | Brand | ID | Excitation Filter | Dichroic mirror | Emission Filter |
---|---|---|---|---|---|---|
1 | DAPI/Hoechst/AMCA | Chroma | Cube set 31000v2 | 350/50x [325-375] | 400LP | 460/50m [435-485] |
2 | FITC/EGFP | Chroma | Cube set 41001 | 480/40x [420-500] | 535/50m [510-560] | |
3 | TRITC/Rhodamine | Chroma | 545/30x [530-560] | 570LP | 620/60m [590-650] | |
4 | Texas Red | Chroma | 560/55x [532-588] | 595LP | 645/75m [607-682] | |
5 | Empty | |||||
6 | Empty |
Detector
Hamamatsu ORCA Flash V2 C11440-22CU CMOS Camera 2048 x 2048 pixels, 16-bit, 30fps at full frame
Camera complete specifications
- Turn on the computer
- Turn on the microscope power bar
If fluorescence is required, turn on the mercury lamp power supply unit (3A) and press ignition (3B)
Mercury lamp must be on for at least 30 min before being turned off and vice-versa
- Log in Windows using your UdM credentials
- Start-up NIS-Elements
The first time you log in the computer, you need to import the microscope settings. To do this follow the instructions Setting-up NIS-Elements
- Save your data
- Close NIS-Elements
- Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
- Get your samples from the microscope
- If used, clean the oil objective with lens cleaner and lens paper
- If fluorescence was used, turn off the mercury lamp power supply unit (3A)
- Turn off the microscope power bar (2)
- Wait until the lamps are cool and cover the microscope
Important Reminders
- Take back your samples including ones in the microscope
- Leave the microscope and the working area clean
- Mercury lamp must be on for at least 30 min before being turned off and vice-versa
- Files can be saved temporarily (during acquisition) on the local C: drive (desktop)
- At the end of each session, copy your data to your external drive and delete it from the local C: drive
- You can store your files on the D: drive (Data Storage). If you do, please create a folder per laboratory using the principal investigator last name. Within, create one folder per user (Firstname_Lastname).
In any case, your files should be removed from the C: drive.
This process is required the first time you are using the instrument. You will usually do it during the training session. It can also be performed if something is not working properly right or if you want to refresh the software interface.
This process will delete all experiment protocols and restore the parameters for the microscope.
- Close NIS-Elements
- Wait until the complete closure of NIS-Elements
- Open the folder Desktop\Logiciels
- Open the software NIS Settings Utility
- Click on the Import tab
- Click on Browse
- Navigate to your desktop
- Select the file Nikon-E600 Settings.bin
- Click Select
- Select all items
- Click Import
- Click OK
- Close the NIS Settings
- Open NIS-Elements
Light path
Download the schematics to follow Transmitted light (Bright field, Phase Contrast, Polarized light) and Reflected light (fluorescence) paths.
Manuals
- Adjust focus drive jitter
- Add a holder for polarized light analyzer
- Nikon Preventive Maintenance
- Adjustment and cleaning Stage Nosepiece, condenser, filter turret, focus drive, shutters
- 100x slightly damaged but not the lens
- Fluorescence cubes damaged: FITC TexasRed
- Focus drive has a jitter
- Replacement mercury lamp bulb HBO 1003W/2
- 256 GB SSD added in workstation for OS
- Windows 10 Installation
- BIOS updated to v2.47
- Camera Firmware updated to v2.11
- NIS-Elements Basic Research v4.6 64-bits installed
- Creation NIS Elements Settings
- FITC and TxRed cubes excitation and emission filters slightly damaged. Dichroic OK
Stand
- Nikon Ti2-E inverted Serial 540156 System 170110-Sys-006287
Light sources
- Transmitted LED light
- ND32 filter
- Manual Polarizer
- Lumencor SpectraX 6-NII-SE Serial 9409 Filters B G R NT V
R 640\30-25
- G 470\24-25
- B 440\20-25
- NT 510\25-25
- V 395\25-25
- GY : 550\15-25
- Storage GY 575\25-25
Condenser
- Motorized condenser
Lens LWD NA 0.52
Filter turret 7 motorized positions
Empty
- Empty
Ph1
Ph2
Shutter
Empty
- DIC N1
Objectives
- 10x/0.3 Plan Fluor Ph1 DLL WD 16 Air
- 40x/0.75 Plan Fluor Ph2 DLL WD 0.72 Air with PF40 Wollaston prism
- 100x/1.3 Plan Fluor Ph3 DLL WD 0.2 Oil with PF/PA 100 Oil Wollaston prism
- Empty
- Empty
- Empty
Position | Name | Brand | ID | Magnification | Numerical Aperture | Immersion | Type | Working distance (mm) | Transmittance (% [nm]) | Technique | Cover glass thickness (mm) |
---|---|---|---|---|---|---|---|---|---|---|---|
1 | 10x/0.3 Air | Nikon | 10x/0.3 Air Plan Fluor Ph1 DLL | 10x | 0.3 | Air | Plan Fluor | 16 | >75% [400-800] | BF, Pol, PhC, Fluo | 0.17 |
2 | 40x/0.75 Air | Nikon | 40x/0.75 Air Plan Fluor Ph2 DLL | 40x | 0.75 | Air | Plan Fluor | 0.72 | >80% [400-750] | BF, Pol, PhC, Fluo | 0.17 |
3 | 100x/1.3 Oil | Nikon | 100x/1.3 Oil Plan Fluor Ph3 DLL | 100x | 1.3 | Oil | Plan Fluor | 0.2 | >75% [400-800] | BF, Pol, PhC, Fluo | 0.17 |
4 | Empty | ||||||||||
5 | Empty | ||||||||||
6 | Empty |
Stage
- Motorized stage Ti2 SHU compatible Serial 127808
- Remote control joystick Ti2-S-JS Serial 127976
- Inserts
- Combo slide and 3cm dish with tilt adjustment insert
- Multiwell plate insert
Filters
- Ex 383-408 DAPI-U DM 425 BA 435-485
- Semrock 96372 M349727 17
- Semrock 96376 M351081 8
- 77074160 Custom Quad C182279 Polychroic and quad bandpass emitter for use with the following single bandpass filters: ET395/25x, ET470/24x, ET550/15x, ET640/30x
- DIC Analyser Ti2CDICACL
- 7707\4656 CFP\YFP\mCherry XT C197767
Position | Cube name | Brand | ID | Excitation Filter | Dichroic mirror | Emission Filter |
---|---|---|---|---|---|---|
1 | DAPI/Hoechst/AMCA | Chroma | Cube set 31000v2 | 350/50x [325-375] | 400LP | 460/50m [435-485] |
2 | FITC/EGFP | Chroma | Cube set 41001 | 480/40x [420-500] | 535/50m [510-560] | |
3 | TRITC/Rhodamine | Chroma | 545/30x [530-560] | 570LP | 620/60m [590-650] | |
4 | Texas Red | Chroma | 560/55x [532-588] | 595LP | 645/75m [607-682] | |
5 | Empty | |||||
6 | Empty |
Detector
Hamamatsu ORCA Flash V2 C11440-22CU Serial 101 081
Workstation
- HP Z440 Workstation
- Intel Xeon E5-1630 v3 @ 3.7GHz
- RAM 32 GB DDR4 2133 MHz (4 x 8 GB)
- OS 256GB SSD 550 MBs
- 2TB HD Data Storage 150 MBs
- Video Card NVIDIA Quadro K620 2 GB DDR3 dedicated memory
- Monitor HP Z24i display 24' 1920x1200
Incubation
- Okolab BoldLine Temperature unit Serial 284-1058 H101 T Unit BL
- Okolab BoldLine CO2/O2 Unit 0-10/1-18 Serial 088-1102
- Okolab OkoTouch Serial 118-224
Consumables
- CO2 Tank
- N2 Tank
Liquid forming in the incubation chamber
This happens when the mixed-gas humidifier is overfilled. Bubbles created by the gas going through the humidification bring liquid into the gas feed line.
- Turn off the Okolab module
- Remove your sample and store it properly
- Dry the incubation chamber with a clean tissue
- Carefully remove the cap of the humidifier glass bottle
The humidifier bottle is made of glass and is very fragile. Pay extra care when manipulating the humidifier bottle
- Remove distilled water from the humidifier
Humidifier shouldn't be more than 2/3rd filled
- Close the humidifier by replacing the cap
- Turn on the Okolab module
Can I use this microscope to look at cell in a dish?
- Yes. This is an inverted microscope designed to look at specimen in a dish or a multi-well plate as well as slides between a glass slide and a 0.17mm thick cover slip.
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