H301-OLYMPUS-IX3-SVR / Issued
Contents
1. 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 www oko lab com sie H301 OLYMPUS IX3 SVR User Manual November 2014 BD Falcon 35x10 Willco 35x10 Corning 35x10 mm MatTek P35G X14 X eS Ibidi p Dish 50 mm low MatTek P50G X30 X Q BD Falcon 60x10 Grainer Petri 60x15 Corning 60x15 mm Q Willco 60x15 jol Figure 13 Ring selection scheme for 60 mm dish www oko lab com page LO H301 OLYMPUS IX3 SVR User Manual November 2014 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 NOTE Magnetic locks are included with sample holder Figure 15 Magnetic lock for 1 x 2 chamber slide To lock the slide push simultaneously the buttons indicated with 3 1 www oko lab com H301 OLYMPUS IX3 SVR User Manual November 2014 8 Working with MW Plates Magnetic locks and Chamber ris er Magnetic locks hold MW plates in place into sample holder illustrated in Figure 16 NOTE Magnetic locks must be ordered separately NOT INCLUDED Product codes MW LOCK 22 5 and MW LOCK 16 5 Magnetic lock MW 22 5 LOCK To be ordered separately Typically used for 6 12 24 48 MW 0 MW 16 5 LOCK To be ordered se2. Nunc and Greiner T25 flask For 3 1 x3 chamber slide 2 Lab Tek Il 1 x2 chambered cover glass For 2 35 mm petri dishes holder Figure 2 Available sample holders www oko lab com Page H301 OLYMPUS IX3 SVR User Manual November 2014 3 Available Lids The following heated glass Lids are available for H301 OLYMPUS IX3 SVR H301 SLIDING LID For easy sample loading and pipetting Included in code H301 OLYMPUS IX3 SVR H301 KOEHLER LID Reduces chamber height to 22 mm and allows imaging under Koehler illumination H301 INJECTION LID 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 Heated glass lid with safety switch Connects to laser controller and automati cally turns laser off when lid is lifted Compatible only with sample holder 1x35 OPTIONAL not included with H301 OLYMPUS IX3 SVR 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 with and without chamber riser Figure 4 illustrates how to remove Sliding Lid when a different 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
3. H301 OLYMPUS IX3 SVR User Manual November 2014 H301 OLYMPUS IX3 SVR Compatible with the following XY stage Olympus IX3 SVR www oko lab com Compatible with the following Okolab Controllers H301 T BL PLUS UNO COMBINED CONTROLLER H401 T DUAL Page 1 H301 OLYMPUS IX3 SVR User Manual November 2014 Index 1 Components and GIMENSIONS cecictsciesdeiateriaretesieees interes este eee aed 3 De Sat PleTIOVGeN Ss x tect ete coerce E E T tee ees eet ec eae ets 3 2 1 Available Sample HolderS essessessossossoesosossosesoesseosessesoesoesoesossoesoesoesoesossoesoesossoosossossossoosoo 3 de AVEDA E 5 J ouae Dee E NE E E E E EE E en E N E EE E EE EEE er maT 5 S22 Koehler Eidos a a a a 6 3o ecn O a E E E E are rsserateee ars Graiere Gee mrs Gears Gears Gers Geers cea cesees 6 34 i mei g lel eh sn er ee ee 7 4 Insertion of the Sample Feedback Temperature SenSor sesseceseecesoecescoeceseecesoecesoecesoecesoecesoececeeoo 8 5 INSertion Of Sample Holder into CNAME siciecios we scis esc arose esrareibrsiere Soars a aa 8 6 Working with 35 or 60 mm Petri Dish Spacing Rings and Magnetic LOCKS ccccceecccccccccscccesccceccs 9 7 Working with 1x3 and 1x2 chamber slides Magnetic lOCKS cece ccc ce nec cceccccecccceccceecccseccceeccs 11 8 Working with MW Plates Magnetic locks and Chamber riser cccccccccccccccccccccccccecccecccecccecccecees 12 9 Connect
4. are available NOTE Please contact info oko lab com if you cannot find the sample holder you are looking for We are constantly adding new inserts to the list 1 35mm Petri dish 1 1x3in chamber slide 1 Lab Tek 1in x2in chambered cover glass 1 Lab Tek II 1in x2in chambered cover glass 1 60mm Petri dish 1 Nunc and Greiner T25 flask 2 35mm Petri dish 2 1x3in chamber slides 2 Lab Tek 1in x2in chambered cover glass www oko lab com be H301 OLYMPUS IX3 SVR User Manual November 2014 2 Lab Tek II 1in x2in chambered cover glass 2 60mm Petri dish 4 35mm Petri dish 1 1x3in chamber slide and 2 35mm Petri dish 1 Lab Tek 1in x2in chambered cover glass and 2 35mm Petri dish 1 Lab Tek II 1in x2in chambered cover glass and 2 35mm Petri dish 1 Lab Tek 1in x2in chambered cover glass and 1 60mm Petri dish 1 Lab Tek II 1in x2in chambered cover glass and 1 60mm Petri dish 1 1x3in chamber slide and 1 60mm Petri dish 1 60mm Petri dish and 1 35mm Petri dish 6 well plates one MW LOCK required 12 well plates one MW LOCK required 24 well plates one MW LOCK required 24 well NUNC GREINER plates one MW LOCK required 48 well plates one MW LOCK required 96 well plates one MW LOCK required It includes the insert for oil immer sion imaging Magnetic lock to hold the standard MW plates in the chamber Magnetic lock to hold the low profile MW plates in the chamber gt Fo
5. hamber with its adapter 1 Follow the arrows shown in Figure 20 Images 1 and 2 for placing the captive screws in the proper housings 2 Place the chamber on the adapter as shown in Figure 20 Image 3 3 Usea 1 5mm metric Allen Wrench to tighten two of the four captive screws in place A in Image 4 while keeping the sliding lid closed See Figure 20 Image 4 www oko lab com bape 3 H301 OLYMPUS IX3 SVR User Manual November 2014 5 Image 6 of Figure 20 shows the chamber fixed on the adapter Captive Screw Figure 20 How to assemble the chamber to the adapter www oko lab com 4 Tighten the remaining two captive screws A in Image 5 while keeping the chamber sliding lid open page 14 H301 OLYMPUS IX3 SVR User Manual November 2014 12 Connection of the Chamber with XY stage Once you have assembled the chamber with adapter you can place it on the XY stage following the steps shown in Figure 21 1 Place the adapter on the stage See Image 1 of Figure 21 2 Follow the instructions shown in Image 2 of Figure 21 for placing socket head cap screws use the provided Allen wrench for tightening the socket head cap screws and fastening the adapter on the stage Screws housings are in dicated with letter A in Image 2 of Figure 21 Screw M3x6 T Figure 21 How to fix the H301 OLYMPUS IX3 SVR to the XY stage www oko lab com a j 5
6. ion of the Gas Supply swiss ssiacisccsesciecctacsseseesesiescvceseaseseasesdesesceecseaceeess ened veseaceseaseeeas tenes 13 10 Working with PENTUSION es vec cvinadvaseces esa esas wesswenswess wens wesa wens wese wens ia E a e ea araia 13 11 Connecting the Chamber to the adapter sssesesesesesesescesececececesescescesecececesesescscecececesesesesessecese 13 12 Connection of the Chamber with XY stage sesesceceseecesoecesoecesoecesoecesoecesoecesoeceseecesoeceseeceseeceo 15 www oko lab com es H301 OLYMPUS IX3 SVR User Manual November 2014 4 Components and dimensions H301 OLYMPUS 1IX3 SVR includes the following components Chamber main body with embedded temperature sensor It connects on the stage with screws Sliding heated glass lid with Indium Tin Oxide ITO conductive coating and embedded temperature sensor Chamber riser It is a removable frame increasing the height of the chamber from 27 to 33 mm often re quired when using multi well MW plates Distance between focal plane 33 and upper side of the glass gt lid with the chamber riser 0 27 istance between focal plane and upper side of the glass gt lid without the chamber riser 0 Sample Holder Order Separately Chamber Base Chamber Riser O Chamber top view gt 155 Figure 1 H301 OLYMPUS IX3 SVR Components and Dimensions 2 Sample Holders 2 1 Available Sample Holders The following sample holders
7. 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 Peep ie ei eg and upper side of the Peep ie ei lid with the chamber riser Distance between focal plane 27 and upper side of the glass lid without the chamber riser 0 Chamber top view gt gt Figure 3 H301 OLYMPUS IX3 SVR Sliding Lid comes as standard www oko lab com H301 OLYMPUS IX3 SVR User Manual November 2014 Sliding Lid 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 22 mm and allowing imaging under Koehler il lumination The Koehler Lid is placed onto the chamber NO SCREWS or TOOLS are necessary Figure 5 shows chamber dimensions with Koehler lid with and without chamber riser H301 KOEHLER LID Distance between focal Pepe aie aeta and upper side of the Pepe aie aeta lid with the chamber riser Distance between focal plane and upper side of the glass lid without the chamber riser 0 a m fo 3 Te j ge lt D y 109 70 Figure 5 H301 OLYMPUS IX3 SVR H301 KOEHLER LID 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 i
8. parately Typically used for 96 MW Figure 16 Magnetic locks for MW plates To be ordered separately Chamber Riser increases chamber s height and is REQUIRED with 6 12 24 48 MW plates regardless of the Lid em ployed Figure 17 shows how to install and fasten the chamber riser Screws location is indicated by letter A Chamber Riser Figure 17 Riser Assembly www oko lab com el 2 H301 OLYMPUS IX3 SVR User Manual November 2014 9 Connection of the Gas Supply A single silicon tubing carries output gas from the Okolab Gas Controller to the H301 OLYMPUS IX3 SVR Silicon tubing connects to a gas input brass opening located on a corner of the H301 OLYMPUS IX3 SVR See Figure 18 Connect by gently pushing silicon tubing onto brass opening Figure 18 Connection with gas supply 10 Working with Perfusion The Chamber Riser included with H301 OLYMPUS IX3 SVR features 12 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 M3x6 Remove small screws as necessary before introducing perfusion tubing Figure 19 shows location of perfusion holes Perfusion dedicated opening 1 Frontal view 2 3D view Figure 19 Perfusion 11 Connecting the Chamber to the adapter Follow the steps shown in the images of Figure 20 and listed below in order to correctly connect the c
9. r 1 1 Pr Se lid d 4x35 M 2x60 M GS60 M 1x60 M or 22 enanos aiceann For 4 35 mm petri dishes For 2 60 mm petri dishes For 1 60mm petri dish and EOLA SO EN petiidish 2 35mm petri dishes 1x35 M 6035 M 1xGS M 2xGS M For 1 35 mm petri dish For 2 35 mm petri dishes and For 1 1 x3 chamber slide For 2 1 x3 chamber slides 1 1 x3 chamber slide gt MW OIL For oil immersion 1 60mm petri dish With all types of well plates z A 6MW 12MW 24MW 24 NUNC MW 48MW For 1 6 well plates holder For 1 12 well plates holder For 1 24 well plates holder For 1 24NUNC GREINER well plates For 1 48 well plates holder holder 96MW LABTEK 35 M LABTEK 60 M 1xLABTEK M 2xLABTEK M For 1 96 well plates holder For 1 Lab Tek 1 x2 chambered For 1 Lab Tek 1 x2 chambered For 1 Lab Tek 1 x2 chambered For 2 Lab Tek 1 x2 chambered cover glass and 2 35mm petri dishes cover glass and 1 60mm petri cover glass cover glass 2 dish lt 1xGS 2xLABTEK M LABTEK II 35 M LABTEK II 60 M 1xLABTEK II M 2xLABTEK II M For 1 1 x3 chamber slide and For 1 Lab Tek Il 1 x2 chambered For 1 Lab Tek II 1 x2 chambered For 1 Lab Tek II For 2 Lab Tek 2 Lab Tek 1 x2 chambered cover glass cover glass and 2 35mm petri dishes cover glass and 1 60mm petri dish 1 x2 chambered cover glass _1 x2 chambered cover glass gt lt gt 1xGS 2xLABTEKII M 1xT25 M 3xGS M For 1 1 x3 chamber slide and 2x35 M For 1
10. s placed onto the chamber NO SCREWS or TOOLS are necessary Figure 6 shows chamber dimensions with the Injection Lid with and without chamber riser www oko lab com s H301 OLYMPUS IX3 SVR User Manual November 2014 Distance between focal Pepe nie ae t and upper side of the Pepe nie ae t H301 INJECTION LID lid with the chamber riser Distance between focal plane and upper side of the glass lid without the chamber riser Chamber Chamber top view _ view gt Figure 6 H301 OLYMPUS IX3 SVR 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 with and without chamber riser H301 LASER INTERLOCK LID Distance between focal plane and upper side of the glass lid with the chamber riser Distance between focal plane and upper side of the glass lid without the chamber riser Chamber top view gt 109 70 Figure 7 H301 OLYMPUS IX3 SVR H301 LASER INTERLOCK LID www oko lab com ew H301 OLYMPUS IX3 SVR User Manual November 2014 4 Insertion of the Sample Feedback Temperature Sensor Insert the Sample Feedback Temperature Sensor through the dedica
11. ted opening located in the H301 OLYMPUS IX3 SVR see Figure 8 Front and 3D views Temperature Sensor dedicated opening Temperature Sensor dedicated opening 1 Frontal view 2 3D view 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 www oko lab com pase H301 OLYMPUS IX3 SVR User Manual November 2014 6 Working with 35 or 60 mm Petri D Magnetic Locks Magnetic locks prevent movement of 35 and 60 mm dishes inside the sample holder ish Spacing Rings and Figure 10 illustrates the available magnetic locks for 35 and 60 mm dishes Threaded magnetic posts allow adjusting holder s height Figure 10 Magnetic locks for 35 and 60 mm dish NOTE Magnetic locks are included with sample holder Spacing rings accounting for variability in dish diameter from one manufacturer to another are also included Figure 11 shows how to position the ring Ring selection guide diagrams rings for 35 and 60 mm dish Figure 12 and Figure 13 respectively
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