H301-K-FRAME / Issued
Contents
1. 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 Psp ei es and upper side of the Psp 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 K FRAME Sliding Lid comes as standard www oko lab com H301 K FRAME User Manual September 2014 Screws M2x4 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 ie ei and upper side of the Pepe ie ei lid with the chamber riser Distance between focal plane and upper side of the glass lid without the chamber riser 0 a gt w 3 om oO ae lt oO y 109 70 Figure 5 H301 K FRAME H301 KOEHLER LID 3 3 Injection Lid The Injection Lid is a heated glass lid with two small openings sealed with fle2. MW LOCK 16 5 MW 22 5 LOCK To be ordered separately Typically used for 6 12 24 48 MW j MW 16 5 LOCK To be ordered separately 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 mount and screw the chamber riser Screws location is indicated by letter A Chamber Riser Figure 17 Riser Assembly www oko lab com el 2 H301 K FRAME User Manual September 2014 9 Connection of the Gas Supply A single silicon tubing carries output gas from the Okolab Gas Controller to the H301 K FRAME Silicon tubing connects to a gas input brass opening located on a corner of the H301 K FRAME 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 K FRAME 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
3. diagrams rings for 35 and 60 mm dish Figure 12 and Figure 13 respectively 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 K FRAME User Manual September 2014 2 BD Falcon 35x10 Willco 35x10 2 Figure 12 Ring selection scheme for 35 mm dish Grainer Petri35x10 Ibidi Dish 35 mm low e e Corning 35x10 mm MatTek P35G X14 X Corning 60x15 mm www oko lab com Figure 13 Ring selection scheme for 60 mm dish Ibidi p Dish 50 mm low MatTek P50G X30 X jol Grainer Petri 60x15 Willco 60x15 page LO H301 K FRAME User Manual September 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 K FRAME User Manual September 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
4. 5 Chamber Riser Chamber Base Figure 1 H301 K FRAME Components and Dimensions 2 Sample Holders 2 1 Available Sample Holders The following sample holders 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 K FRAME User Manual September 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
5. Figure 19 Perfusion www oko lab com bape 3 H301 K FRAME User Manual September 2014 e 11 Connection of the Chamber with XY stage H301 K FRAME fits on the XY Stage without any screws To fix the chamber on the stage follow the steps shown in Figure 20 1 Place the chamber on the stage See Image 1 of Figure 20 2 Incline the chamber and slide it in direction of the springs until it snaps into the housing See Image 2 of Figure 20 3 Image 3 of Figure 20 shows the chamber fitted on the XY Stage Figure 20 Connection of the Chamber with XY Stage www oko lab com page 14
6. H301 K FRAME User Manual September 2014 H301 K FRAME Compatible with the following XY stages ASI all models including z piezo Leica all models with 160x110 opening Ludl all models with 160x110 opening Marzhauser all models Olympus all models with 160x110 opening Prior H107 Zeiss all models with 160x110 opening Compatible with the following Okolab Controllers H301 T BL PLUS UNO COMBINED CONTROLLER H401 T DUAL www oko lab com Page 1 H301 K FRAME User Manual September 2014 Index Me Component and CNS SIONS eranen E E donee cence edesencueneueueueneucdseecausscucueshuseedeue soe 3 L Same RTE ga ere Pee ee ane EP ene E Pee tT eT eC ee ee en ere 3 2 1 Available Sample HolderS ssossossossossoesosoososessesseosessesoesoesoesossoesoesoesoesossoesoosossoosossoosoosoosoo 3 SB MVANADIC Lids ssssssiriseri rrira a S seca seas secanearsccaeaee sas 5 Li SANS UC en ee eee 5 Cy KOENE CIT eee ere er Mere rare er en ORT Ore ve ee eee ee er ee err tren 6 Io MSC CIO agai aetitte acs vee ate vette vse ters setae da EE eta then seta bins eenseens eens 6 sa Eiser INGCTIOC L1G 4 inawcadanniuna dann iune a tunaneus toneneen a 7 4 Insertion of the Sample Feedback Temperature SeENsor ccccccccccccccccccccccccccccecccecccecccecccecceeccsecs 8 5 Insertion of Sample Holder into Chamber sesesessesesececesesesescecececececesesescoesecececesesesceces
7. esececeseseseoo 8 6 Working with 35 or 60 mm Petri Dish Spacing Rings and Magnetic LOCKS ccccceecccccccccscccesccceeccs 9 7 Working with 1x3 and 1x2 chamber slides Magnetic lOCKS cc cce ccc ceecccceccccncccceccceecccescccesces 11 8 Working with MW Plates Magnetic locks and Chamber riser cccccccccccccccccccccecccecccecccecccecccecees 12 8 Connection OF the Gas SUDDUWY sacie seco cess poe ace nee bw e nee eee auw ie EEEE A EEEE EEA EERE 13 10 Working with Pel ISlOM ie cece coc cacencuwcnuecsuccaseceuacerece vec vacevecevaucweneceoneweneuions seca ceneuesacessuesacossoocasioes 13 11 Connection of the Chamber with XY stage ssesescesesoecesoecesoecesoecesoecesoecesoecesoeceseecesoeceseeceseeceo 14 es www oko lab com H301 K FRAME User Manual September 2014 1 Components and dimensions H301 K FRAME 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 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 Sliding Lid Distance between focal plane 33 and upper side of the glass gt lid with the chamber riser 0 Distance between focal plane 27 and upper side of the glass lid without the chamber riser 0 Sample Holder Order Separately 200 8
8. 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 GS35 M For 1 1 x3 chamber slide and 2 35mm petri dishes 4x35 M 2x60 M GS60 M 1x60 M For 4 35 mm petri dishes For 2 60 mm petri dishes For 1 60mm petri dish and Sa petiidsh 1 1 x3 chamber slide MW OIL For oil immersion 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 60mm petri dish With all types of well plates 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 Il 1 x2 chambered For 1 Lab Tek Il For 2 Lab Tek 2 Lab Tek 1 x2 chambered cover glass cover glass and 2 35mm petri dishes cover glass and 1 60mm pe
9. on of the Sample Feedback Temperature Sensor Insert the Sample Feedback Temperature Sensor through the dedicated opening located in the H301 K FRAME see Figure 8 Frontal 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 Wa 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 K FRAME User Manual September 2014 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 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
10. tri 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 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 K FRAME User Manual September 2014 3 Available Lids The following heated glass Lids are available for H301 K FRAME H301 SLIDING LID For easy sample loading and pipetting Included in code H301 K FRAME 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 K FRAME 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
11. xible 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 with and without chamber riser www oko lab com s H301 K FRAME User Manual September 2014 Distance between focal Pepe nie ae te and upper side of the Pa nie ae te 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 K FRAME 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 K FRAME H301 LASER INTERLOCK LID www oko lab com ew H301 K FRAME User Manual September 2014 4 Inserti
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