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DH-40i User`s Manual

1. EEE 10 Warner Instruments Inc DH 40i Rev 04 03 02 3 The DH 40i Micro Incubation Chamber from Warner Instruments is designed to provide complete environmental control over RC 40 Series Chambers and 35 mm culture dishes while maintaining the ability to make both short and long term imaging studies This versatile system readily accommodates culture dishes from several manufacturers and will adapt to any microscope stage supported by Warner Instruments Features of the DH 40i include Y Designed for Warner s RC 40 Series glass bottom quick change imaging chambers Y Designed for 35 mm culture dishes Adapts to dishes made by Willco Wells Corning and Falcon v Permits imaging temperature and gas atmosphere control v Unique dish clamps permit either open or closed use v Heated top coverslip optional eliminates condensation when used as a closed chamber Yv Accommodates upright and inverted microscopes Warner Instruments Inc DH 40i Rev 04 03 02 4 INTRODUCTION The DH 40i Culture Dish Incubator is designed to facilitate the imaging of live cells in a RC 40 Series chamber or cell culture dish Cells can be imaged using either an open or closed dish format The open dish format readily accommodates the use of microelectrodes or rapid throughput tools while the closed dish format can be used to perform long term studies or to maintain pH CO2 or other environmental factors T
2. i e T2 can be placed directly in the bath to check solution temperature The temperature control unit regulates the heating current by sensing the temperature of the incubator via the T1 control thermistor in the CC 28 cable Heating of perfusate can be achieved using our SH 27B or SF 28 In Line solution heaters which are also controlled via our TC 324B and TC 344B Temperature Controllers If using 35 mm culture dishes in the open format Warner Instruments RC 33 perfusion chamber inserts are available for Corning dishes and can be used together with the DH 40i These inserts incorporate our diamond fluidics design to maintain laminar perfusate flow Closed Format Applications In the closed format the platform and windowed cover section are used together to maintain an enclosed environment Gas ports inlet and outlet in the cover enable the user to control the cells pH CO2 and O2 The windowed cover is easily removed allowing quick replacement of cell culture dishes during replicate trials The cover secures onto the platform via two magnetic posts Access ports are provided in the cover to permit insertion of accessory lines such as a monitor thermistor into the enclosed chamber In the closed format resistive heating is achieved in the same manner as described for the open format Warner Instruments Inc DH 40i Rev 04 03 02 10 APPENDIX Cleaning and maintenance The DH 40i is constructed of anodized aluminum
3. DH 40i Rev 04 03 02 Warner Instruments Inc Micro Incubation Platform For RC 40 Chambers Model DH 40i Warner Instruments Inc 1125 Dixwell Avenue Hamden CT 06514 800 599 4203 203 776 0664 203 776 1278 fax 7 Warner Instruments Inc DH 40i Rev 04 03 02 Table of Contents PESCRIFTION e 3 INTRODUCTION c 4 COMPONENTS 22 20 22 5 LEE EE NE NN AA 5 rS 5 FA CCOSS OF NOS c M 5 SETUL Ran NEE SEELE EEE EEE EEE EEE EEE IE EISEN 6 General C mn 6 Open Formal L EERRAEEe 6 NN 6 PN 6 TCG EEE RENNE EEE NE EE 6 Pu 7 Closed OT m E E E EE NN 7 100 2 SPREE NE EEE NE 7 Cover 7 Nr 8 TT SEERE EE ee 8 TINN 8 OPERATION c O 9 Open Format Appl CATIONS vvs 9 Closed Format Applications uu uue 9 ATTENDI a HH N 10 Cleaning and maintenance nn aaa 10 Warranty and service ee TERR STA SES AVE UTE ER ben en gehe Teen YVES ba UNS 10 Az avec 10 a TT
4. It can be easily cleaned with common laboratory detergents or with ethyl alcohol Warranty and service Service We recommend that all questions regarding service be referred to our Technical Support Department Normal business hours are 8 30 AM to 5 00 PM EST Monday through Friday Our offices are located at Warner Instruments 1125 Dixwell Avenue Hamden CT 06514 800 599 4203 203 776 0664 203 776 1278 fax support warneronline com http www warneronline com contact html Warranty The DH 40i is covered by our Warranty to be free from defects in materials and workmanship for a period of one year from the date of shipment If a failure occurs within this period we will either repair or replace the faulty component s This warranty does not cover failure or damage caused by physical abuse In the event that repairs are necessary shipping charges to the factory are the customer s responsibility Return charges will be paid by Warner Instruments 7 Warner Instruments Inc
5. closed and has a handle to simplify manipulation Two perfusion tubes input and aspiration are incorporated into the platform cover and represent the input and output flow lines for the chamber The input tube has a smaller diameter and is open at the tip The aspiration tube can be identified by its larger 7 Warner Instruments Inc DH 40i Rev 04 03 02 8 diameter and has a plugged tip Both tubes are fully adjustable removable are bent to aid in positioning and will maintain their orientation when the cover is removed and replaced Finally there are two magnets embedded into cover to provide a secure attachment between the cover and the platform base Cover assembly l Begin by placing a clean 40 mm round coverslip into the recess on the cover Secure the coverslip using the supplied o ring 2 Attach the gas line to the gas input port on the cover 3 Attach the inflow and outflow perfusion lines to their respective perfusion tubes 4 If needed carefully route any accessory lines through the miscellaneous entry ports on the cover Heating Follow the heating instructions for the open bath configuration detailed on page 6 Placement 1 Place your chamber into the platform and secure the cover into position over the chamber 2 With the cover in place adjust the position of the perfusion tubes into the chamber Begin solution flow Make adjustments to 1 the position of the perfusion tubes and 2 the
6. e side of the platform See figure on top of next page for thermistor placement A small amount of mineral oil can facilitate good thermal contact between the feedback thermistor and the metal of the platform 7 Warner Instruments Inc DH 40i Rev 04 03 02 7 Slip the blued colored leads over the two resistive heating elements and then plug the CC 28 cable into the temperature controller Consult the temperature controller user s manual for methods of maintaining the temperature of the clamped culture dish Placement If imaging place the fully assembled DH 40i into your microscope s DH 40i Stage Adapter and mount onto the microscope Closed Format Chambers Chambers are assembled into the platform as described on page 6 Cover components The DH 40i cover has a gas entry port notches for the dish clamps and heater blocks two access ports and two perfusion tubes for solution exchange An inverted cover is shown to the right The gas entry port connects to your atmospheric control system Gas vents into the chamber around the window which provides good distribution while eliminating window condensation The dish clamp and heater block notches are provided for the obvious reasons They also serve to orient the cover onto the platform The access ports can be used to provide a vent for waste gasses or to provide easy entry for special lines of your choosing Each entry port swivels open and
7. emperature control is available in both open and closed formats RC 40 Series Glass Bottom Quick Change Chambers are small volume imaging chambers featuring rapid solution exchange short working distances and an open bath Since bath volumes are generally small exchange times are measured in seconds even when flow rates are less than 1 ml min The DH 40i incubator is primarily designed to work with the RC 40 Series chambers but will work equally well with 35 mm glass bottom cell culture dishes from Willco Wells Corning or Falcon The incubator is machined in two interlocking sections and metal adapters are supplied to insure a secure fit for supported 35 mm culture dishes 7 Warner Instruments Inc DH 40i Rev 04 03 02 o COMPONENTS The completed DH 40i assembly is comprised of a platform a Series 40 chamber or 35 mm culture dish with adapter ring if necessary and a cover The assembly is mounted onto a microscope using a Warner microscope stage adapter Thermal control is provided using a CC 28 cable and temperature controller Platform The platform directly accepts a Series 40 chamber Supported culture dishes are positioned using one of four supplied adapter rings see Accessories The platform also includes two resistive heating elements a 19 mm diameter aperture dish clamps and a single hole for attaching a control thermistor e g via Warner s CC 28 cable The periphery of the platform is framed wit
8. h stainless steel plates for mounting magnetic holders when the DH 40i is used in the open configuration Cover The cover shown inverted is used to form an enclosed micro incubation system The cover window is sealed using a 40 mm round cover glass The window cover glass is positioned via a recessed annular lip and is held in place using a rubber o ring When in place the cover glass seals a 36 mm widow useful for illumination purposes Accessories Four colored metal adapter rings are included to insure a close fit between the platform and a cell culture dish Rings are tapered for a precise fit and should be mounted with the notch facing down A special tool i e a modified 18G hypodermic needle is provided to assist in removing adapter rings from the Bottom Section Color codes for the different adapter rings are shown below Adapter ring color codes Willco Wells Models D3512 and D3522 Red Falcon 37 mm Falcon 38 mm Corning 25000 series 7 Warner Instruments Inc DH 40i Rev 04 03 02 6 SETUP General Comments The DH 40i is quickly and easily assembled The platform may be used alone for open format applications while the cover is required for closed format applications The cover contains several ports allowing entry of atmospheric and perfusion lines as well as electrode and thermistor access We recommend the use of the RC 40 Series quick change chambers when using the DH 40i in clo
9. perfusion flow to achieve the desired flow characteristics Stop solution flow until the DH 40i is placed onto the microscope 3 Ifimaging place the assembled DH 40i into your microscope s DH 40i Stage Adapter and mount onto the microscope 4 Begin gas and solution delivery 5 Adjust the entry ports for gas venting if necessary Chamber with cover removed Chamber assembled 7 Warner Instruments Inc DH 40i Rev 04 03 02 9 OPERATION Open Format Applications Only the platform together with one of the four colored adapter rings and the dish clamps is required for open format use The open format enables easy access for microelectrodes perfusion lines and aspiration lines Cell culture dishes can be used with and without covers and dish perfusion can be simplified by use of Warner s RC 40 Series Perfusion Chambers The open format permits imaging using either upright or inverted microscopes Warner Series 40 Stage Adapters are used to mount the DH 40i onto most commonly used microscope stages Contact our Tech Support Department for custom stage adapters Resistive heating of the DH 40i is achieved using Warner s TC 324B or TC 344B Temperature Controller and a CC 28 cable The two blue CC 28 connectors attach directly to each resistive heating element Feedback control is provided by thermistor i e Tl which is inserted into the platform s metal chassis via a special hole The CC 28 cable s bath thermistor
10. sed format Open Format Chambers If using an RC 40 Series chamber begin by assembling the chamber Use of an adapter ring is not necessary If using a cell culture dish begin by selecting the appropriate adapter ring for your dish See adapter guide page 5 Place the adapter ring notch side down on a firm surface and insert the cell culture dish into the ring by gently pushing it into place The chamber is placed into the recess in the heater block platform and is clamped into place Dish clamps are provided and are placed into two small holes located on dish clamp either side of the platform These holes can be found directly adjacent to the heater blocks See the figure to en Y magnetic the right for clamp positioning c E holder Perfusion An input tube and an aspiration tube are provided with the DH 40i to facilitate perfusion within the chamber These tubes can be found as part of the cover assembly The aspiration tube can be identified by its larger diameter It also has a plug at its tip The input and aspiration tubes can be easily removed from the cover and inserted into the provided magnetic holders for placement onto the steel shelves of the platform Attach perfusion lines to the input and aspiration tubes and place into position Heating If heating with Warner s TC 324B or TC 344B temperature controllers slip the white colored T1 feedback thermistor on the CC 28 cable into the milled hole in th

DH-40i User`s Manual

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