Okolab H301-KEYENCE-BZX User manual

User Manual
H301-KEYENCE-BZX
IST 2792_REV01

Index
1CHAMBER COMPATIBILITY 2
2COMPONENTS AND DIMENSIONS 2
1.1 AVAILABLE SAMPLE HOLDER............................................................................................................................. 3
3INSTALLATION 4
1.2 INSTALLATION OF THE CHAMBER INTO THE XY STAGE ......................................................................................... 4
1.3 INSERTION OF THE SAMPLE HOLDER INTO CHAMBER BASE................................................................................... 5
1.4 WORKING WITH 35 OR 60 MM PETRI DISH –SPACING RINGS AND MAGNETIC LOCKS............................................. 6
1.5 WORKING WITH 1X3’’ AND 1X2’’ CHAMBER SLIDES -MAGNETIC LOCKS................................................................... 8
1.6 WORKING WITH MW PLATES -MAGNETIC LOCKS AND CHAMBER RISER................................................................. 9
4GAS CONNECTION 10
5WORKING WITH PERFUSION 11
6INSERTION OF THE SAMPLE FEEDBACK TEMPERATURE SENSOR 12
7CLEANING 13
8FIGURE LIST 13
9DOCUMENT REVISION TABLE 13
1Chamber Compatibility
H301-KEYENCE-BZX is a heated top stage chamber designed for Keyence BZX systems with XY stage
having 160x110mm opening. It can be configured with both BOLD LINE and UNO controllers
2Components and Dimensions
H301-KEYENCE-BZX includes the following components:
●Lid with embedded temperature sensors, to place on the chamber body (No screws or tools are
necessary).
Riser. It is a removable frame increasing the height of the chamber from 22,65 to 28,65 mm, often
required when using multi-well (MW) plates.
●Sample Holder.
●Chamber Base.

Figure 1. H301-KEYENCE-BZX –Components and Dimensions.
1.1 Available Sample Holder
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
#1 35mm Petri-dish
1xGS-M
#1 1x3in. chamber slide
1xLABTEK-M
#1 Lab-Tek 1in.x2in. chambered cover glass
1xLABTEK-II-M
#1 Lab-Tek II 1in.x2in. chambered cover glass
1x60-M
#1 60mm Petri-dish
1xT25-M
#1 Nunc and Greiner T25 flask
2x35-M
#2 35mm Petri-dish
2xGS-M
#2 1x3in. chamber slides
2xLABTEK-M
#2 Lab-Tek 1in.x2in. chambered cover glass
2xLABTEK-II-M
#2 Lab-Tek II 1in.x2in. chambered cover glass
2x60-M
#2 60mm Petri-dish
4x35-M
#4 35mm Petri-dish
GS35-M
#1 1x3in. chamber slide and #2 35mm Petri-dish
LABTEK-35-M
#1 Lab-Tek 1in.x2in. chambered cover glass and #2 35mm Petri-dish
LABTEK-II-35-M
#1 Lab-Tek II 1in.x2in. chambered cover glass and #2 35mm Petri-dish
LABTEK-60-M
#1 Lab-Tek 1in.x2in. chambered cover glass and #1 60mm Petri-dish
LABTEK-II-60-M
#1 Lab-Tek II 1in.x2in. chambered cover glass and #1 60mm Petri-dish

GS60-M
#1 1x3in. chamber slide and #1 60mm Petri-dish
6035-M
#1 60mm Petri-dish and #1 35mm Petri-dish
6MW
6-well plates
12MW
12-well plates
24MW
24-well plates
24MW-NUNC
24-well NUNC/GREINER plates
48MW
48-well plates
96MW
Holder to image 96-well plates with LD objectives.
MW-OIL
Holder to image 6/12/24/48/96-well plates with immersion objectives.
Figure 2. Available Sample Holders.
3Installation
1.2 Installation of the Chamber into the XY Stage
H301-KEYENCE-BZX fits into the XY stage of the Keyence BZX systems in the same way as Keyence
stage holders.

Figure 3. Connection of the Chamber with XY Stage
1.3 Insertion of the Sample Holder into Chamber Base
Sample holders fit into the chamber base and are held in place by magnets embedded within both the
chamber and the holder. 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 4.

Figure 4. Introduction of the Sample Holder inside the Chamber Base.
1.4 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 5 illustrates the available magnetic locks for 35 and 60 mm dishes. Threaded magnetic posts
allow adjusting the holder's height.
Magnetic Locks for 35 and 60 mm petri dishes.
Included in the corresponding sample holder
Figure 5. Magnetic Locks for 35 and 60mm dish.
Note ►Magnetic locks are included with sample holders.
Spacing rings accounting for variability in dish diameter from one manufacturer to another are also
included. Figure 6 shows how to position the ring. Ring selection guide diagrams: rings for 35 and 60 mm dish
-Figure 7 and Figure 8, respectively.

1
3
2
Figure 6. 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 holders.
Note ►Spacing rings do not block the petri inside its accommodation, but they are designed to center the petri with
respect to the observation circular window.
BD Falcon-35x10 Willco-35x10
Grainer-Petri35x10 Ibidi μ-Dish 35 mm-low
MatTek-P35G-X14-XCorning-35x10 mm
Figure 7. Ring selection scheme for 35 mm dish.

Corning-60x15 mm
BD Falcon-60x10
Ibidi μ-Dish 50 mm-low
Grainer - Petri 60x15
MatTek-P50G-X30-X
Willco 60x15
Figure 8. Ring selection scheme for 60 mm dish.
1.5 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.
Note ►Magnetic locks are included with sample holders.
1
2
Figure 9. Magnetic lock for 1'' x 3'' chamber slide.

1
2
3
Figure 10. Magnetic lock for 1'' x 2'' chamber slide. To lock the slide, push simultaneously the buttons indicated with (3).
1.6 Working with MW Plates - Magnetic locks and Chamber riser
Magnetic locks hold MW plates in place into sample holder illustrated in Figure 11
Figure 11. Magnetic locks for MW plates. Ships with 6, 12, 24, 48 MW plates.
The Chamber Riser increases the chamber's height and it is required to accommodate 6, 12, 24, 48
MW plates.
Figure 12 shows how to mount and screw the chamber riser. Screws location is indicated by letter A.

Figure 12. Riser Assembly.
4Gas Connection
A single silicon tubing carries output gas from the Okolab Gas Controller to the H301-KEYENCE-BZX.
Silicon tubing connects to a gas input - brass opening - located on a side of the H301-KEYENCE-BZX. See
Figure 13.
Figure 13. Gas Input Port.

Figure 14. Output Gas Connection to H301-KEYENCE-BZX.
5Working with perfusion
The H301-KEYENCE-BZX features 23 perfusion holes (10 in the chamber riser, 13 in the chamber base)
on the sides for the insertion of perfusion tubing up to 2.5 mm in outer diameter. Small screws plug the
perfusion holes when not in use (screws plug M3x0.5 THD). Remove small screws plug as necessary before
introducing perfusion tubing.
Figure 15 shows location of perfusion holes.

Figure 15. Perfusion.
6Insertion of the Sample Feedback Temperature Sensor
Insert the Sample Feedback Temperature Sensor through the dedicated opening located in the H301-
KEYENCE-BZX (see Figure 16).
To read the temperature of the culture medium and realize a self-calibration of the chamber, fix the
Temperature Sensor tip with adhesive tape on the bottom of the dish inside the H301-KEYENCE-BZX. For more
information about the self-calibration, please refer to the User Manual of the Okolab Controller in use.
Figure 16. Insertion of the temperature sensor inside the chamber.

7Cleaning
●Turn the system off and pull the mains plug out the socket
●Wait 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 directly 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.
8Figure List
Figure 1. H301-KEYENCE-BZX –Components and Dimensions. .......................................................................... 3
Figure 2. Available Sample Holders......................................................................................................................... 4
Figure 3. Connection of the Chamber with XY Stage ............................................................................................ 5
Figure 4. Introduction of the Sample Holder inside the Chamber Base................................................................ 6
Figure 5. Magnetic Locks for 35 and 60mm dish................................................................................................... 6
Figure 6. 1) Insertion of the ring, 2) insertion of the 35 mm dish, 3) insertion of the magnetic lock. ................. 7
Figure 7. Ring selection scheme for 35 mm dish. .................................................................................................. 7
Figure 8. Ring selection scheme for 60 mm dish. .................................................................................................. 8
Figure 9. Magnetic lock for 1'' x 3'' chamber slide. ................................................................................................ 8
Figure 10. Magnetic lock for 1'' x 2'' chamber slide. To lock the slide, push simultaneously the buttons
indicated with (3)...................................................................................................................................................... 9
Figure 11. Magnetic locks for MW plates. Ships with 6, 12, 24, 48 MW plates. ................................................... 9
Figure 12. Riser Assembly. .................................................................................................................................... 10
Figure 13. Gas Input Port. ...................................................................................................................................... 10
Figure 14. Output Gas Connection to H301-KEYENCE-BZX................................................................................. 11
Figure 15. Perfusion. .............................................................................................................................................. 12
Figure 16. Insertion of the temperature sensor inside the chamber. .................................................................. 12
9Document Revision Table
Revision Number
Additions or changes
Date
01
First edited
April 2022
IST 2792_REV01