Okolab H301-OLYMPUS-IX-SUSP User manual

H301-OLYMPUS-IX-SUSP User Manual
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H301-OLYMPUS-IX-SUSP
Compatible with the following XY stages
Compatible with the following Okolab Controllers
•Olympus: IX-SUSP, GX-SVR, GX-SFR, BX3-SSU,
IX2-SFR, IX2-SVL-2
•H301-T-BL-PLUS
•UNO-COMBINED-CONTROLLER
•H401-T-DUAL

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Index
1. Components and dimensions................................................................................................... 3
2. Sample Holders................................................................................................................... 3
2.1 Available Sample Holders....................................................................................................... 3
3. Available Lids ..................................................................................................................... 4
3.1 Sliding lid .......................................................................................................................... 5
3.2 Koehler Lid........................................................................................................................ 5
3.3 Injection Lid ...................................................................................................................... 6
3.4 Laser Interlock Lid............................................................................................................... 7
4. Insertion of the Sample Feedback Temperature Sensor.................................................................. 8
5. Insertion of Sample Holder into Chamber ................................................................................... 8
6. Working with 35 or 60 mm Petri Dish –Spacing Rings and Magnetic Locks ........................................... 9
7. Working with 1x3’’ and 1x2’’ chamber slides - magnetic locks........................................................10
8. Connecting the Gas Supply and Filling the Humidity Module ...........................................................12
9. Working with Perfusion ........................................................................................................12
10. Connection of the Chamber with XY stage .................................................................................13
11. Cleaning ..........................................................................................................................14

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1. Components and dimensions
H301-OLYMPUS-IX-SUSP includes the following components:
•Chamber main body with embedded temperature sensor
•Sliding heated glass lid with Indium Tin Oxide (ITO) conductive coating and embedded temperature sensor
•Integrated humidity module. It is not necessary to select an additional humidity module
Distance between focal plane
and upper side of the glass lid
Chamber top view
Sliding Lid
Chamber Base
Sample Holder-Order Separately
Integrated Humidty Module
Figure 1. H301-OLYMPUS-IX-SUSP - Components and Dimensions.
2. Sample Holders
2.1 Available Sample Holders
The following sample holders are available.
NOTE: Please contact [email protected] if you cannot find the sample holder you are looking for. We are constantly
adding new inserts to the list.
1x35-M single
#1 35mm Petri-dish
1xGS-M single
#1 1x3in. chamber slide
1xLABTEK-M single
#1 Lab-Tek 1in.x2in. chambered cover glass
1xLABTEK-II-M single
#1 Lab-Tek II 1in.x2in. chambered cover glass
1x60-M single
#1 60mm Petri-dish

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1x35-M single
For #1 35mm Petri-dish
1x60-M single
For #1 60mm Petri-dish
1xGS-M single
For #1 1’’x3’’ chamber slide
1xLABTEK-M single
For #1 Lab-Tek 1’’x2’’ chambered cover glass
1xLABTEK-II-M single
For #1 Lab-Tek II 1’’x2’’ chambered cover glass
Figure 2.Available sample holders.
3. Available Lids
The following heated glass Lids are available for H301-OLYMPUS-IX-SUSP:
•H301-SLIDING-LID: For easy sample loading and pipetting –Included in code H301-OLYMPUS-IX-SUSP
•H301-KOEHLER-LID-2*: Reduces chamber height to 23 mm and allows imaging under Koehler illumination
•H301-INJECTION-LID-2*: Heated glass lid with two small openings (sealed with flexible plastic) allowing injec-
tion or permanent access to the sample. Compatible only with sample holder 1x35
•H301-LASER-INTERLOCK-LID-2*: Heated glass lid with safety switch. Connects to laser controller and auto-
matically turns laser off when lid is lifted. Compatible only with sample holder 1x35
* OPTIONAL –not included with H301-OLYMPUS-IX-SUSP

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3.1 Sliding lid
The Sliding Lid is a heated glass lid allowing for easy sample loading and pipetting. The Sliding Lid is screwed onto the
chamber.
Figure 3 shows chamber dimensions with the Sliding Lid. Figure 4 illustrates how to remove Sliding Lid, when a differ-
ent Lid is necessary.
NOTE: Sliding Lid MUST BE REMOVED when using any other Lid. The Sliding Lid is fixed onto the chamber with 4
screws. Screws location is indicated by letter A in Figure 4 (image 2 and 3). Keep Sliding Lid fully closed to access
screws labeled A in image 2. Open Sliding Lid to access screws labeled A in image 3.
Distance between focal plane
and upper side of the glass lid
Chamber top view
Sliding Lid
Figure 3. H301-OLYMPUS-IX-SUSP (Sliding Lid comes as standard)
1 2 3
Sliding Lid
A
A
Screws M2x6
Figure 4. Assembly of the sliding lid
3.2 Koehler Lid
The Koehler Lid is a heated glass lid reducing the chamber’s height to 23 mm and allowing imaging under Koehler il-
lumination. The Koehler Lid is placed onto the chamber (NO SCREWS or TOOLS are necessary).

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Figure 5 shows chamber dimensions with Koehler lid.
Distance between focal plane
and upper side of the glass lid
Chamber top view
H301-KOHLER LID-2
Figure 5.H301-OLYMPUS-IX-SUSP + H301-KOEHLER-LID-2
3.3 Injection Lid
The Injection Lid is a heated glass lid with two small openings sealed with flexible plastic allowing injection or perma-
nent access to the sample. NOTE: Compatible with sample holder 1x35 ONLY. The Injection Lid is placed onto the
chamber (NO SCREWS or TOOLS are necessary).
Figure 6 shows chamber dimensions with the Injection Lid .

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Distance between focal plane
and upper side of the glass lid
Chamber top view
H301-INJECTION LID-2
Figure 6. H301-OLYMPUS-IX-SUSP + H301-INJECTION-LID
3.4 Laser Interlock Lid
The Laser Interlock Lid is a heated glass lid with a safety switch connecting to the laser controller. It automatically
turns the laser off when the lid is lifted. NOTE: Compatible with sample holder 1x35 ONLY. The Laser Interlock Lid is
placed onto the chamber (NO SCREWS or TOOLS are necessary).
Figure 7 shows chamber dimensions with the Laser Interlock Lid.
Distance between focal plane
and upper side of the glass lid
Chamber top view
H301-LASER-INTERLOCK LID-2
Figure 7. H301-OLYMPUS-IX-SUSP + H301-LASER-INTERLOCK-LID

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4. Insertion of the Sample Feedback Temperature Sensor
Insert the Sample Feedback Temperature Sensor through the dedicated opening located in the H301-OLYMPUS-IX-SUSP
(see Figure 8, Frontal and 3D views).
1.Frontal view 2.3D view
Temperature Sensor dedicated opening
Temperature Sensor dedicated opening
Figure 8. Insertion of the temperature sensor inside the chamber.
5. Insertion of Sample Holder into Chamber
Sample holders fit into the chamber base and are held in place by magnets embedded within both chamber and hold-
er. To introduce a sample holder with the proper orientation, match the red dot on the holder to the one on the
chamber base, as illustrated in Figure 9.
Insert the adapter by
superimposing the red points
Figure 9. Introduction of the Specimen Holder inside the Chamber Base.

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6. Working with 35 or 60 mm Petri Dish –Spacing Rings and
Magnetic Locks
Magnetic locks prevent movement of 35 and 60 mm dishes inside the sample holder.
Figure 10 illustrates the available magnetic locks for 35 and 60 mm dishes. Threaded magnetic posts allow adjusting
holder’s height.
Magnetic Locks for 35 and 60 mm petri dishes.
Included in the corresponding sample holder
Figure 10.Magnetic locks for 35 and 60 mm dish
NOTE: Magnetic locks are included with sample holder.
Spacing rings with variable diameter are also included in according to the needs of the user. Figure 11 shows how to
position the ring inside the sample holder. Ring selection guide diagrams: rings for 35 and 60 mm dish -Figure 12 and
Figure 13, respectively.
1
3
2
Figure 11. 1) Insertion of the ring, 2) insertion of the 35 mm dish, 3) insertion of the magnetic lock.
NOTE: Spacing rings are included with sample holder.

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Grainer-Petri35x10 Ibidi μ-Dish35mm-low
Corning-35x10mm MatTek-P35G-X-14-X
Willco-35x10BD Falcon-35x10
Figure 12. Ring selection scheme for 35 mm dish.
MatTek-P50G-X-30-X
Grainer - Petri 60x15
Willco60x15
BD Falcon 60x15
Ibidi μ-dish 50mm-low
Corning-60x15mm
Figure 13. Ring selection scheme for 60 mm dish.
7. Working with 1x3’’ and 1x2’’ chamber slides - magnetic
locks
Magnetic locks prevent movement of 1’’x 3’’ and 1’’x 2’’ chamber slides inside of the sample holder.

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NOTE: Magnetic locks are included with sample holder.
2
1
Figure 14. Magnetic lock for 1'' x 3'' chamber slide.
1
3
2
Figure 15. Magnetic lock for 1'' x 2'' chamber slide. To lock the slide, push simultaneously the buttons indicated with (3).

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8. Connecting the Gas Supply and Filling the Humidity Module
A single silicon tubing carries output gas from the Okolab Gas Controller to the H301-OLYMPUS-IX-SUSP. Silicon tubing
connects to a gas input - brass opening - located on a side of the H301-OLYMPUS-IX-SUSP. See Figure 16. Connect by
gently pushing silicon tubing onto brass opening.
H301-OLYMPUS-IX-SUSP includes an integrated humidity module in order to maintain a high humidity percentage inside
the chamber. Fill the humidity module inside the chamber through the water input - brass opening - located on a side
of the H301-OLYMPUS-IX-SUSP (See Figure 16) using a silicon tubing 2 mm ID and a syringe*.
*NOTE: Do not exceed 0.05 l
Gas Input
Water Input
Figure 16. Connection with gas supply.
9. Working with Perfusion
H301-OLYMPUS-IX-SUSP features 8 perfusion holes for the insertion of perfusion tubing up to 2.5 mm in outer diameter.
Small screws plug the perfusion holes when not in use. (Grub screws M3x3). Remove small screws as necessary before
introducing perfusion tubing.
Figure 17 shows location of perfusion holes.
1. Frontal view 2. 3D view
Perfusion dedicated opening
Figure 17. Perfusion

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10. Connection of the Chamber with XY stage
Follow the steps shown in the images Figure and listed below in order to correctly connect the chamber with XY stage.
1. Place the chamber on the stage and tighten 4 captive screws (See Figure Image 1 and Image 2). Captive screws
housings are indicated with letter B in Figure Image 2
2. Use a 3mm metric Allen Wrench to tighten the four captive screws while keeping the chamber sliding lid open
(See Figure Image 2)
Captive Screws
A
B
1 2
3
Figure 18. Connection of the Chamber with XY Stage

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11. Cleaning
•Turn the system off and pull the mains plug out the socket
•Remove the lid from the chamber main body, and keep it separate from the chamber main body while the
chamber cools down.
•To clean the body and the glass lid of the chamber, wipe with a soft micro-fiber cloth. For stubborn smudges,
you can damp the soft micro-fiber cloth with ethyl alcohol (product code UN1170). Do not put any liquid di-
rectly on the chamber. While cleaning the glass lid, do not apply strong force to the surface of the glass lid
because it can be damaged.
H301-OLYMPUS-IX-SUSP_User Manual Rev 02_Published in January 2020