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icon	build
title	Français
type	primary
url	https://wiki.umontreal.ca/display/Microscopie/Nikon + Ti2-E

Tabs Container

id	Instrument Tabs
title	Nikon Ti2-E
direction	horizontal

Tabs Page

id	Description
title	Description

Nikon Ti2-E inverted

Oko

Nikon Ti2-Oko microscope

Roger Gaudry Building, Room R-619
Advanced Microscope Tier 1 usage price
Instrument awarded to Dr Paradis-Bleau the Canadian Foundation for Innovation (CFI #34495)

Advanced Microscope Tier 1 usage price

in 2015

Applications

Inverted microscope
Widefield imaging
- Brightfield

Applications

Transmitted light, Bright-field

- Phase contrast
- Polarized light, DIC
- Fluorescence
Live imaging

Time-lapse imaging

Timelapse imaging
Incubation

Image Added

Tabs Container

direction	horizontal

Tabs Page

title	Description

Description

Light sources

LED lamp for transmitted light
Lumencor SpectraX for fluorescence

Expand

title

Lumencor SpectraX complete

Complete specifications

Source

ID

Filter

type

	Excitation

wavelengths

(nm)

Compatible f

Dluorophores	Nominal power (mW)	Measured Power at sample (mW)
Violet	395/25

Bandpass

[382-407]	DAPI, Hoechst	295	59.4
Blue	440/20

Bandpass

	[430-450]	CFP	256	90.1
Cyan	470/24

Bandpass

	[458-482]	FITC, GFP	196	49.8
Teal	510/25

Bandpass

[497-522]	YFP	62	15.8
Green/Yellow	550/15

Bandpass

	[542-557]	TRITC, Cy3	260	69.9
Green/Yellow (Storage)	575/25

Bandpass

	[562-587]	mCherry	310	78.2
Red	640/30

Bandpass

[625-655]

Cy5

231

Objectives

20x/0.5 Air Ph1 WD 2.1

60x/1.4 Oil DIC WD 0.13

100x/1.45 Oil Ph3 WD 0.13

44.8

View file

name	Lumencor_SpectraX_Brochure.pdf
height	250

View file

name	Lumencor_SpectraX_Filter recommandation.pdf
height	250

View file

name	Lumencor_SpectraX_Manual.pdf
height	250

Objectives

100x/1.45 Oil DIC WD 0.13

4x/0.2 Air

WD 20

20x/0.5 Air Ph1
20x/0.75 Air DIC

WD 1.0

60x/1.4 Oil DIC
100x/1.45 Oil Ph3
100x/1.45 Oil DIC

Expand

title

Objectives complete

Complete specifications

Position	Name	Brand	Full name	Identifier	Magnification	Numerical aperture	Immersion	Type	Working distance (mm)	Transmittance (% [nm])

Techniques

Applications	Cover glass thickness (mm)
1

20x

4x/0.

5

2
Air

Ph1

Nikon

20x

4x/0.

5 Air Plan Fluor Ph1MRH102012.1

2
Plan Apo Lambda

MRD00045

4x

0.2

Air

Plan Apo Lambda

20

>80% [400-

750

1000]

BF

, Pol, PhC

2
, Fluo	0.17

60x

20x/

1.4 Oil DIC

0.5
Air Ph1

Nikon

60x

20x/

1.4 Oil Plan Apo Lambda DIC N2MRD016050.13

0.5 Ph1
Plan Fluor

MRH10201

20x

0.5

Air

Plan Fluor

2.1

>80% [400-750]
Measured 72%

>80% [475-725]

BF, Pol,

DIC

3
PhC, Fluo	0.17

100x

20x/

1.45 Oil Ph3

0.75
Air DIC

Nikon

100x

20x/

1.45 Oil

0.75 DIC
Plan Apo Lambda

Ph3MRD31905

DIC N2

MRD00205

20x

0.75

Air

Plan Apo Lambda

1.0

0.13

>80% [

475

400-

750

950]
Measured 88%

BF, Pol,

PhC

4
DIC, Fluo	0.17

100x

60x/1.

45

4
Oil DIC

Nikon

100x

60x/1.

45

4 Oil

Plan

Plan Apo Lambda
DIC N2

MRD01905

MRD01605

0.

60x

1.4

Oil

Plan Apo Lambda

0.13

>80% [475-

750

5
725] Measured 28%	BF, Pol, DIC, Fluo	0.17

4x

100x/

0.2 Air

1.45
Oil Ph3

Nikon

4x

100x/

0.2 Air

1.45 Oil
Plan Apo Lambda

MRD00045

Ph3

MRD31905

100x

1.45

Oil

Plan Apo Lambda

0.13

20

>80% [

400

475-

1000

6
750] Measured 9%	BF, Pol, PhC, Fluo	0.17

20x

100x/

0.75 Air

1.45
Oil DIC

Nikon

20x

100x/

0.75 Air

1.45 Oil
Plan Apo Lambda
DIC N2

MRD00205

MRD01905

100x

1.45

Oil

Plan Apo Lambda

0.13

>80% [

400

475-

950

750]
Measured 11%

BF, Pol, DIC, Fluo

0.17

Filters

DAPI
GFP/FITC (CFP)
Cy5
DAPI/GFP/Cy3/Cy5 (requires Cy3 filter in Lumencor SpectraX Green/Yellow position)
DIC Analyzer
CFP/YFP/mCherry (requires mCherry filter in Lumencor SpectraX Green/Yellow position)

Expand

title

Filters complete

Complete specifications

Position	Name	Brand	ID	Excitation

Filter

	Dichroic

mirror

	Emission

Filter

	Comments
1	DAPI	Nikon	DAPI-U HQ	395/25x [383-408] 303258	425LP	460/50m [435-485] 307914	C-FL-C DAPI-U HQ
2	GFP	Semrock	GFP-4050B-000	466/40x [446-486]	495LP	525/50m [500-550]	Nikon ID 96372
3	Cy5	Semrock	Cy5-5070A

617

618/

55x

50x
[

590

5-

645

6]

652LP

697

698/

77m

70
[

659

6-

736

6]

Nikon

ID 96376

ID 96376 M351081
4	DAPI/GFP/Cy3/Cy5	Semrock	C182279	None	409/493/573/652	432/515/595/681	77074160 Custom Quad C182279. Excitation filters are in the Lumencor SpectraX light source
5	DIC Analyzer	Nikon	Ti2-C-DICACL	Not Applicable	Not Applicable	Not Applicable	Used for DIC imaging
6	CFP\YFP\mCherry	Semrock	C1997767	None	459/526/596	475/543/702	77074656 Excitation filters are in the Lumencor SpectraX light source

Detector

Hamamatsu ORCA Flash 4.0 V2 C11440-22CU

CMOS Monochrome Camera 2048 x 2048 pixels, 16-bit, 30 images/s at full frame
Tabs Page

id	User Guide
title	User Guide

UI Expand

expanded	true
title	Startup

Turn on the computer (#1)

Remove the cover from the microscope

Turn on the microscope power bar (#2)

If incubation is required, turn on the Okolab incubation module (#3A), the Lauda water bath (#3B) and open the CO₂ (#3C) and N₂ (#3D) tanks

Warning
Make sure the humidifier and the water bath are clean and properly filled with distilled water.

Use your UdeM credentials to log in to Windows

Start the software NIS-Elements

Info
The first time you use the instrument, you need to import the microscope settings into the software. To do this follow the instructions First use protocol.

Expand

title	Complete specifications

Camera	Hamamatsu ORCA Flash 4.0 V2
Sensor Type	CMOS
Sensor Category	Monochrome
Nb Pixels	4.2 M
Pixel Layout	2048 x 2048
Pixel size	6.5 um
Sensor size	13.312 mm x 13.312 mm
Sensor diameter	mm
Bit depth	16-bit
Speed at full resolution	30 images/s
Max QE	83 %
Readout noise	1.6 e⁻
Cooling	-10 C Air
Dark Current	0.06 e⁻/pixel/sec
Full well capacity	30 000 e-
Dynamic Range	1: 37 000
Interface	USB 3.0
Mount	C-mount

Image Added

View file

name	Hamamatsu_Orca Flash 4.0_V2_C11440-22CU_Instruction Manual.pdf
height	250

View file

name	Hamamatsu_Orca Flash 4.0_V2_C11440-22CU_Technical Note.pdf
height	250

Tabs Page

title	User Guide

User Guide

UI Expand

expanded	true
title	Startup

If not already done, turn on the computer (#1) and log in to Windows using your UdeM credentials
Remove the dust cover from the microscope
Turn on the microscope power bar (#2) on the desk between the microscope and the computer
if incubation is required, turn on Okolab incubation module (#3A), the Lauda water bath (#3B) and open the CO₂ (#3C) and N₂ (#3D) cylinders near the door
Warning
Make sure the humidifier and water bath are properly filled with distilled water.
When using the instrument for the first time, it is necessary to import the microscope configuration into the software. See the First Use section below before starting the software
Start NIS-Elements

UI Expand

title	First Use

When using the instrument for the first time, it is necessary to import the microscope configuration into the software. This procedure is usually carried out during the training session. However, it can also be used to reset the software if it does not display correctly, for example.

Warning
Running this procedure will erase all your experiment protocols and reset the software to its original settings. If you are not sure, ask for support.

If NIS-Elements is open, close it and wait until it has completely shut down (this may take up to 30 seconds)
On the Desktop, open the Softwares folder
Open NIS Settings Utility
Click on the Import tab
Click on Browse
Navigate to your C:\Users\Public\Documents
Select the file Nikon_Ti2-Oko_NIS Settings.bin
Click Select
Select all items
Click Import
Click OK
Close the NIS Settings Utility
You can now reopen NIS-Elements

UI Expand

title	Loading samples

During this procedure, you will:

Set the microscope in a safe configuration
Load your sample
Find and adjust the focus

Once completed, your sample will be ready for acquisition.

UI Expand

title	Initial Focus

On the microscope, gently push the transmitted light arm backward.
In NIS-Elements, click Escape to move the objectives to a safe position.

Click on the lowest magnification objective.

Info

The lowest magnification is the safest to use due to its long working distance: the sample will appear in focus well before the objective lens gets close to it. It is recommended to always perform the initial focusing with the lowest magnification objective. Since the objectives are parfocal, focusing with these objectives will make it easier to locate the sample when switching to higher magnification objectives.

Place the test slide on the microscope stage, with the coverslip toward the objective.
Note
Using a test slide will significantly reduce the time needed to set up the instrument.
If necessary, use the joystick to move the stage so the sample is centered under the objective.
Gently return the transmitted light arm to the vertical position.
In NIS-Elements, click Escape again to return the objectives to their normal position.
Click on the desired optical configuration (BF, DAPI, GFP, etc.).
Click Live (») to activate the light and display the camera image.
Adjust the light intensity and exposure time to obtain a well-exposed image.
Adjust the focus until the image is perfectly sharp.
Click Stop to stop the Live or on Off to turn off the light

Tip
The focus is around Z = 2100 µm. The Z-position value is displayed on the microscope remote control screen.

UI Expand

title	Secondary focus

Warning
First perform the initial focusing with the safest objective before selecting a higher-magnification objective.

UI Expand

title	Focusing with air objectives

After completing the initial focus:

In NIS-Elements, click on the desired objective (20x PhC or 20x DIC)
Info
The 20x DIC objective is the best air objective because it has the highest numerical aperture (0.75).
Click on the desired optical configuration (BF, DAPI, GFP, etc.).
Click Live (») to activate the illumination and display the camera image on the screen.
Adjust the light intensity and exposure time to obtain a properly exposed image.
Adjust the focus using the precision knob until the image is perfectly sharp.
Click Stop to stop the Live mode or click on the optical configuration Off to turn off the illumination.
Click Escape to move the objectives to a safe position.
On the microscope, you can remove the test slide and install your sample.
In NIS-Elements, click Escape once again to return the objectives to their normal position.

Your sample is now ready for acquisition!

UI Expand

title	Focusing with oil objectives

After completing the initial focusing:

In NIS-Elements, click Escape to move the objectives to a safe position.
Click on the 63x, 100x DIC, or 100x PhC objective to select the desired lens.
Info
The 100x DIC objective is the best oil immersion lens because it has the highest numerical aperture (1.45).

Place a single drop of oil on the objective.
Click Escape again to return the objectives to their normal position.
Click on the desired optical configuration (BF, DAPI, GFP, etc.).
Click Live (») to activate the illumination and display the camera image on the screen.
Adjust the light intensity and exposure time to obtain a properly exposed image.
Adjust the focus using the precision knob until the image is perfectly sharp.
Click Stop to stop the Live mode or click on the optical configuration Off to turn off the illumination.

Your sample is now ready for acquisition!

UI Expand

title	Shutdown

Save your data

Close NIS-Elements software

Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive

Turn off the computer

If oil objectives were used, clean it with lens cleaner and lens paper (not Kimwipes)

If incubation was used, turn off the Okolab incubation module (#3A),Lauda water bath (#3B) and close the CO₂ (#3C) and N₂ (#3D) tanks

Wait until the SpectaX has cooled down and turn off the microscope power bar (#2)

Cover the microscope

Note

title	Important Reminders

Collect your samples, especially those in the microscope

Leave the microscope and workspace clean

UI Expand

title	Storage management

Files can be saved temporarily (during acquisition)

to elete

on the local C: drive (desktop)
At the end of each session, copy your data to your external drive and

d

delete it from the local C: drive
You can store your files on the D: drive (Data Storage). If you do, please create

one

a folder per laboratory using the principal investigator

's

last name.

Inside

Within, create

a

one folder per user

using the following nomenclature (First Name_Last Name

(Firstname_Lastname).

Note

title

Important

In any case,

do not store

your files

on

should be removed from the C: drive.

UI Expand

title

First use AnchorFirstUseFirstUseWhen using the microscope for the first time, you need to import the microscope settings into the software. You will usually do this during the training session.
This procedure can also be performed if something is not working properly and if you want to reset the software to its original settings.

Note
This process will delete all experiment protocols and reset the software to the original settings for this specific microscope.

If open, close NIS-Elements and wait until it is completely closed (up to 30 seconds)
On your Desktop open the Softwares folder
Open NIS Settings Utility
Click on the Import tab
Click on Browse
Navigate to your Desktop
Select the file Nikon-Ti2 Settings.bin
Click Select
Select all items
Click Import
Click OK
Close the NIS Settings
You can now reopen NIS-Elements

Tabs Page

id	Light path
title	Light path

The following schematics depict the light path for transmitted (bright-field and Phase Contrast) and reflected (fluorescence) lights.

PDF

name	LightPath_Nikon Ti2.pdf

Tabs Page

id	Manuals
title	Manuals

Available manuals

Tabs Page

id	Log
title	Log

UI Expand

expanded	true
title	To do

Check condenser filters if labels match

Check why 20x/0.75 DIC objective has no DIC prism

Condenser DIC prism is DIC N1 while objective requires N2

Check why at 4x Condenser gives a black shadow on the righ side of the image

Adjust Objective XYZ offset

Check plane because FOV is not flat

Shutdown

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
If used, clean oil objectives with lens cleaner and paper
Select the lowest magnification objective and press escape to place the objectives in a safe position
If used, turn off the Okolab incubation module (#3A),Lauda water bath (#3B) and close the CO₂ (#3C) and N₂ (#3D) cylinders
Wait until the SpectaX fan is off and then turn off the microscope power bar (#2)
Turn off the computer
Cover the instrument with the protective dust cover

Note
Take back your samples including ones in the microscope Leave the microscope and the working area clean

Tabs Page

title	Log

Log

UI Expand

expanded	true
title	To do

Condenser DIC prism is DIC N1 while objective requires N2
Check why at 4x Condenser gives a black shadow on the righ side of the image
NIS Objective Z offset
Nitrogen Tank replacement Praxair 1066 106723504
CO2 Tank replacement Praxair 1013 106722901

UI Expand

title	2025-10-01 Installation Win 11 23H2

Migrated to Windows 11

UI Expand

title	2025-09-03 Fixing bent stage insert

Fixed Stage insert bent
Cleaning all objective and filter cubes
Added XY Offset

UI Expand

title	2025-08-30 Incubation maintenance

Fixed incubator tubing misplaced
Added better attachment for tubing on the incubation chamber
Checked incubation chamber for obstruction

UI Expand

title	2024-03-20 Incubation maintenance

Fixed incubator tubing misplaced
Added better attachment for tubing on the incubation chamber
Checked incubation chamber for obstruction

UI Expand

expandedtrue

title	2023-11-29 Incubation maintenance

Fixed condenser holder loose
20x/0.5 Ph1 dirty or damaged likely dirty with varnished

UI Expand

title	2022-09-22 Direct connection with Nikon processing workstation

Added network cable between Acquisition and Processing workstations
Added shortcuts in Desktop\Documentation

UI Expand

title	2022-03-16 Cleaining objective and fixing stage insert

Clean 60x objective (nail polish on it !)
Fix damaged stage insert
Stage insert tilt adjustment
Objective XY offset adjustments

UI Expand

title	2022-03-16 Added 4x objective

Installed new objective 4x/0.2 Plan Apo Lambda
Updated configuration file

UI Expand

title	2022-02-21 Incubation complete maintenance

Computer maintenance
Imaging test slides
Re-install DCAM API drivers v22.2.6391
Okolab incubation complete maintenance
- Disassemble Okolab stage insert
- Disconnect the pipes
- Disconnect the Lauda water-bath
- Clean everything with 10% acetic acid (excepted the plastic pipes and washers)
- Rinse with regular tap water 2 times
- Rinse with distilled water once
- Dry, reassemble and reconnect
- Refill with the Lauda water-bath with 3L of milliQ water
- Set the Okolab temperature Control Mode to Chamber
- Set the Okolab temperature to 65°C
- Let run for 1h. This will sterilize the water.
- Check for any leakage
- Set the Okolab temperature Control Mode back to Sample
- Set the Okolab temperature back to 37°C

UI Expand

title	2021-12-16 Maintenance

Cleaning objectives
Control for incubation chamber liquid intrusion

UI Expand

title	2021-11-30 CO2 Calibration

CO₂ Calibration for Okolab: Offset of 1%

UI Expand

expanded	true
title	2021-11-05 QC

Control for illumination quality

See Illumination Report_Nikon Ti2_2021-11-05.pdf

See
Liquid light guide adjustment

UI Expand

title	2021-09-16 Objectives setup

20x/0.75 DIC objective was dirty and has been cleaned
Printed and displayed a Memo about available air and oil objectives
Adjustment of camera angle
Objective calibration
Objective XY offset
Updated NIS settings
Added light path schematics to

wikipedia

Wiki

UI Expand

title	2021-09-15 Fix Okolab Temp Probe

Okolab display a NAN message for the free thermal sensor
The probe is made of two wires that need to be in contact to measure the temperature properly
- Turn off the Okolab module
- Disconnect the free sensor probe
- Cut out the damaged part
- Carefully expose the two wires
- Connect the two wires together

UI Expand

title	2021-09-13 Incubation setup

Added label on gas bottles
Added line mark on humidifier

UI Expand

title	2021-07-20 Complete reinstallation

512 GB SSD installed for OS
Windows 10 Installation
BIOS updated to v2.47

Tabs Page

id

title	Technical Datasheet

title

Technical Datasheet

Stand

Nikon Ti2-E inverted Serial 540156 System 170110-Sys-006287

Light sources

Transmitted LED light
- ND32 filter
- IR filter
- Manual Polarizer
Lumencor SpectraX 6-NII-SE Serial 9409

Condenser

Motorized condenser Ti2-C-TC-E Serial 519097
Lens LWD NA 0.52
Filter turret 7 motorized positions
1. Empty
2. Empty
3. Ph1
4. Ph3
5. Shutter
6. Empty
7. DIC N1

Objectives

20x

4x/0.

5 Air Ph1 WD 2.1

60x/1.4 Oil DIC WD 0.13

100x/1.45 Oil Ph3 WD 0.13

100x/1.45 Oil DIC WD 0.13

4x/0.2 Air WD 20

2 Air MRD00045
20x/0.5 Air Ph1 MRH10201
20x/0.75 Air DIC MRD00205
60x/1.4 Oil DIC MRD01605
100x/1.45 Oil Ph3 MRD31905
100x/1.45 Oil DIC MRD01905

20x/0.75 Air DIC WD 1.0

Stage

Motorized stage Ti2 SHU compatible Serial 127808
Remote control joystick Ti2-S-JS Serial 127976
Inserts
- Multi-well plate Ti2-S-HW with tilt adjustment (no incubation)
- Combo slide

3cm

- 3 cm dish Ti2-S-HU with tilt adjustment (no incubation)
- Okolab H101-CellASIC Frame with perfusion ports
  - 1 x 35mm petri dish 1x35-M + cover
  - 2 Chamber Slide 2xGS-M+ cover
  - 1 Multi-well for oil objectives MW-OIL
  - 6-well plate 6MW+ cover
  - 1 CellASIC + cover

Filters

DAPI Cube Ex 383-408 DAPI-U DM 425 BA 435-485
GFP Cube Semrock 96372 M349727 17
Cy5 Cube 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 Ti2-C-DICACL
C197767 7707\4656 CFP\YFP\mCherry XT

Detector

Hamamatsu ORCA Flash V2 C11440-22CU

CMOS Monochrome Camera 2048 x 2048 pixels, 16-bit, 30fps at full resolution

Serial 101081

Workstation

HP Z440 Workstation
I155431-CIB
Intel Xeon E5-1620 v4 @ 3.5GHz
RAM 32

GB DDR4

GB DDR4-2400 1200 MHz ECC (4 x 8 GB)
OS 500 GB

SSD 530

NVMe via PCIe to M2 Adapter 2 500 MB/s
4 TB HD Data Storage (2 x 2 TB spanned volume) 130 MB/s
Video Card nVidia GTX 1080 8GB

DDR5

GDDR5 dedicated memory
Monitor HP Z24i display 24' 1920x1200
Software NIS-Elements AR v5.02

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
Lauda water-bath Model Eco RE415 S LCK 4910 Serial LCK-4910-16-0006 Okolab 1322-1006 2017

-05-30 Tabs PageidFAQ

-05-30

Anti-vibration Table

TMC #63463542-03 Serial 205235

Consumables

CO₂ Tank
N

₂ Tank

Liquid Light Guide

₂ Tank
Liquid Light Guide

Manuals

Tabs Page

title	Troubleshooting & FAQ

Troubleshooting

UI Expand

title	Chamber does not reach the desired temperature (or is very slow to warm up)

This can happen when the liquid running through the chamber is not flowing properly.

Pause your experiment
Turn off the Okolab environment controller 3A and 3B
Disconnect the blue end of the connection pipe (at the junction with the spring shape objective warmer)
Place the open end into a dish to collect liquid
Turn the Lauda water bath back ON (3B)
The liquid should flow quite rapidly, if not proceed as follow
- Turn OFF the Lauda water bath (3B)
- Take the 20 mL syringe located inside the drawer labelled Tubing
- Connect it to the open end of the blue pipe
- Use the syringe to blow pressurized air into the pipes to clear it out
- Remove the syringe (and store it back into the Tubing drawer)
- Test if the liquid is now flowing properly by turning the Lauda water bath back ON (3B)
  - If not repeat the procedure
  - If so, turn OFF the Lauda water-bath (3B)
  - Reconnect the blue and green pipes together
  - Turn the Okolab environment controller 3A and 3B back ON

Warning
Be careful not to touch your sample when performing these actions as it may displace your current acquisition

Info

title	How the Okolab chamber works

The liquid from the water bath enters the lead first via the red pipe
Then it goes through the lead circuitry to keep it warm
It exists via the

unlabeled

unlabelled tube and enters the incubation chamber main body
It goes through the circuitry of the main body and exits via the green labelled tube

One must maintain a decent amount of liquid in the Lauda water bath to avoid bubble formation in the circuitry.
Because the circuitry inside the top lead and the incubation chamber main body is thin it can get clogged. The procedure above can solve this issue.

UI Expand

title	Liquid forming inside the incubation chamber

This happens when the mixed-gas humidifier is overfilled. Bubbles created by the gas going through the humidifier can bring liquid into the gas feed line.

Turn off the Okolab module and the water bath
Remove your sample and store it properly
Dry the incubation chamber with a clean tissue
Carefully remove the cap of the humidifier glass bottle

Warning
Pay extra care when manipulating the humidifier bottle as it is made of glass and is very fragile

Remove distilled water from the humidifier

Info
Humidifier shouldn't be more than 2/3^rd filled

Close the humidifier by replacing the cap
Turn on the Okolab module and the water bath

UI Expand

title	Liquid forming inside the incubation chamber but the humidifier is not overfilled

This can happen during long experiments. The high humidity of the gas mixture condensates in the pipe between the humidifier and the incubation chamber.

Pause your experiment
Disconnect both ends of the yellow tube from the humidifier
Drain the tube from any liquid
Reconnect the yellow tube to the humidifier
You can use a syringe or gas pressure to blow out any liquid from the incubation chamber cover

Warning
Be careful not to touch your sample when performing this action

Wipe any liquid with a tissue
Reconnect the yellow tube to the incubation chamber

FAQ

UI Expand

title	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
The objectives are optimized to image through thin glass bottom multi-well plates
You may also image specimen mounted between a slide and a 0.17mm thick coverslip
For long timelapse, be aware of photo-toxicity

UI Expand

title	How can I warm up the incubation chamber faster?

You may use a trick to

warmup

warm-up the incubation chamber faster. Using this method. you should reach a stable temperature within 30 minutes.

Warning
Do NOT proceed with your sample but with a blank control (dish with distilled water for example)

Place your blank control in the incubation chamber

Immerge

Immerse the tip of the sample temperature probe in the blank
Set the Okolab temperature Control Mode to Chamber (Settings>Temperature>Control Mode>Chamber>Save)
Set the Okolab temperature to 50°C (Home>Temperature>50°C>Set)
Check the temperature of the sample (Menu Magnifier>Sample Temperature). It should take between 10 to 15 minutes for the blank to reach 30°C.

Info
The Lauda water bath is faster to warm up water than to cool it down. Make sure to anticipate and not pass beyond the desired temperature. If your desired temperature is 37°C, you can stop the procedure when the blank has reached 30°C.

When the blank has reach the desired temperature

Set the Okolab temperature back to 37°C (Home>Temperature>37°C>Set)
Set the Okolab temperature Control Mode to Sample (Settings>Temperature>Control Mode>Sample>Save)
Wait 10 to 20 minutes until the temperature stabilizes
Then you can safely replace the blank with your sample

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Nikon Ti2-Oko microscope

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Versions Compared

Old Version 90

New Version Current

Key

Nikon Ti2-Oko microscope

Applications

Description

Light sources

Objectives

4x/0.2 Air

Filters

Detector

User Guide

Log

Technical Datasheet

Stand

Light sources

Condenser

Objectives

Stage

Detector

Workstation

Incubation

Anti-vibration Table

Consumables

Manuals

Troubleshooting

FAQ