Nikon Ti2-E (en)

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]	505LP	535/50m [510-560]
3	TRITC/Rhodamine	Chroma	Cube set 41002c	545/30x [530-560]	570LP	620/60m [590-650]
4	Texas Red	Chroma	Cube set 41004	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.

Light path schematics.pdf

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
1. Empty
2. Empty
3. Ph1
4. Ph2
5. Shutter
6. Empty
7. 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]	505LP	535/50m [510-560]
3	TRITC/Rhodamine	Chroma	Cube set 41002c	545/30x [530-560]	570LP	620/60m [590-650]
4	Texas Red	Chroma	Cube set 41004	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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