Bioptechs Delta TC3 Culture Dish System
Caution: Read carefully before use!

The system is comprised of three main components a temperature controller, a stage adapter and the Delta TC3 dish. It utilizes a new and improved technology which is much faster and more accurate than traditional peripheral stage warmers to simulate host conditions on the stage of your microscope and provide an optimal optical environment for microscopy. The controller has a fast learning curve to anticipate changes in the dish temperature due to perfusion while regulating temperature and compensating for cooling due to surface evaporation.

• Check to make sure there is no visible damage in shipment and save all packing materials.
• Check the packing list and call Bioptechs, Inc. at 724-282-7145 if there are any discrepancies.
• Follow instructions carefully to ensure a thorough understanding of this unique system.

1. Turn the unit on using the main on-off switch located on the rear panel. All further adjustments are made on the front panel.
2. Turn the display switch to the "DISH" position.The temperature display will read a number between 3.7 and -4.2. Press the black button labeled "CALIBRATE" and make sure the display reads 25.0°C. If adjustment is necessary, turn the corresponding screw in the adjustment hole with the enclosed screwdriver. CW=increase, CCW=decrease. Repeat this procedure with the display switch set to REF.
3. Move the display switch to the "SET" position. Insert the enclosed screwdriver into the opening marked "SETPOINT" and set the controller to read a value 2.0°C less than the desired setpoint. Note: The surface probe reads faster than the cell surface because it is not immersed in liquid and is in direct contact with the heating surface. This is to your advantage.
4. Plug in the stage adapter. With the display switch in the DISH position it will read room temperature.
5. You now have a choice. You can either turn the controller off for a minute before loading a dish with cells or leave the controller on and remember to place the MODE switch into the confocal position for a second. Then return it to the AUTO position immediately before the dish is inserted into the stage adapter. This will re-enter the preset value into the control circuit and give the learning curve a head start.
6. Wet the metallic surface of the thermistor located in the aperture of the stage adapter with a light film of immersion oil. Place a dish into the stage adapter and secure it by turning the dish 15° CW.
7. If a dish containing cells is still warm from prior incubation the Delta TC3 will reestablish temperature in about 15 seconds. If the contents of the dish are at room temperature Delta TC3 will require 2 minutes to reach setpoint. The dish should contain about 2ml of media for proper operation. Cells can be then plated directly on the microscope by pipetting into the instantaneously warmed dish. Do not power up with a dry dish.
8. Adjust the SETPOINT by immersing the reference thermistor into the dish and read its temperature by turning the display switch to the "REF"position. During this time we recommend floating some mineral oil on top of the media to eliminate the effects of surface evaporation. Allow the thermistor time to equilibrate before reading its value. Compare the reference temperature to the regulated temperature and make the appropriate adjustments to the SETPOINT value. Alternate method; Place a Bioptechs liquid crystal temperature indicator dot into the dish and adjust the SETPOINT until it turns green.
9. Turn the controller off prior to removing the dish from the stage adapter to prevent the controller from attempting to regulate a nonexistent load. This will cause the learning curve to go beyond reasonable values. If this happens, you can always place the controller into the Confocal mode for a second to reset the learning curve.

Special Notes:
 If you are perfusing* cells and the dish temperature deviates from setpoint more than the 0.9° C error window, an alarm will sound, current will be shut off to the dish and a red light will come on next to the red reset button. To silence the alarm and resume heating, press the red button. This will return the controller to cold startup mode and it will ignore the error window until the temperature again reaches setpoint minus 0.9°C. You can use the confocal switch at any time to reset the the controllers learning curve.

Use of the Delta TC3 in confocal applications:
The control characteristics of the Delta TC3 enable it to almost instantaneously compensate for temperature changes that may occur in the dish due to perfusion, surface evaporation or other ambient conditions. The sensitivity of the controller, 0.1°C, coupled with the speed and intensity of the controller's response, may cause the coverslip to flex a detectable amount during confocal imaging. Therefore, we have added a supplemental circuit which bypasses the dynamic temperature control mode and permits you to temporarily send a constant DC current into the dish to eliminate Z axis displacement of the specimen.
1. Operate the Delta TC3 in the auto (dynamic) mode when you are not scanning or acquiring images.
2. When you intend to acquire an image, place the mode switch (toggle) on the front of the controller in the down position labeled Confocal. This will disable the dynamic operation of the controller and send a stable DC current into the dish. The value of the DC current can be adjusted with the potentiometer labeled Confocal. To adjust the DC value, turn the display switch to "Volts Out" with the mode switch in the "Confocal" position and adjust the Confocal potentiometer. A good starting point is 2.7V shown as 27.0 on the display. Return the display switch to the dish position and check to see that the DC value maintains the setpoint value. Make sure the controller is returned to the auto mode immediately after the image is acquired.
• An optional TTL actuated interface is available to enable a computer controlled transition between automatic and confocal modes. Thus eliminating the need to "babysit" the system.

Continual Delta TC3 Perfusion with the Micro-Perfusion Pump:
The Micro-Perfusion Pump Starter Set is shipped with a dual channel tubing. One of the tubes has a black band and smaller ID. It is made this way so that one of the tubes will have a slower fluid transport rate. Attach the supply line from your media to the the barb on the input side of the black band pump tubing and another tube from the output side of this tube to the input of the dish. Attach tubing from the aspiration tube in the dish to the input side of the other pump tube and a line from the output side of this pump tube to a waste container. Adjust both the ,height of the aspiration tube, in the dish for level control, and pump speed for continuous perfusion. *Bioptechs recommends the Micro-Perfusion Pump.

Stage Adapter
The stage adapter provides support for the dish, electrical contacts and the surface temperature probe. Make sure the adapter is firmly attached to the scope. When mounting a dish make sure there is a film of immersion oil on the surface probe to enhance thermal conductivity. To mount a dish place the dish into the stage adapter by matching the tabs on the dish with the openings in the stage adapter. Then, while applying sufficient downward force so as to compress the contacts in the base of the adapter, rotate the dish 15 degrees CW until it comes to rest under the opposing metal tabs.
• A pair of mounting holes on either side of the dish aperture are provided to attach perfusion devices.
• There is a thermistor on the top of the terminal cover which can be read by setting the display switch
on the controller to the "REF" position.
Note: If corrosion is present on the electrical contacts, it should be cleaned immediately. The contacts can corrode if exposed to media in the presence of an electrical current. Corrosion on stage adapters is preventable and not covered under warranty.
Dishes are removed by rotating the dish 15° CCW then lifting out of the retainer.

Delta TC3 Dishes

Delta TC3 dishes are made of a polystyrene ring which has a specially coated clear glass substrate bonded onto the bottom surface forming a 2ml liquid containment structure having a 35mm OD which tapers down to a 23mm optical aperture. The glass substrate is available in two thicknesses; 0.5mm for low N.A. and #1.5 coverslip for high N.A. applications. In standard production, the 0.5mm thick substrate is bonded to a clear dish ring and shipped with clear lids. The #1.5 coverglass is available bonded to black or clear dish rings and shipped with corresponding lids. All other characteristics of the dishes are identical. We recommend the use of black dishes for fluorescence applications. The thickness of the dish can also be determined from the stock number on the dish package.
04200405 = 0.5mm thick clear
04200415B = #1.5 coverslip thickness black
04200415C = #1.5 coverslip thickness clear
Custom configurations available.

High Numeric Aperture Use:
If you are are using high numeric aperture objectives (immersion) typically with high N.A. dishes, it will be necessary to regulate the temperature of the objective as well. This is due to the fact that the optical coupling medium, oil, glycerine or water, also has a thermal coupling effect. In this case a Bioptechs, Inc. Objective Heater and an Objective Heater Controller will be necessary for uniform temperature across the field.

Objective Heater Description:
The Bioptechs Objective Heater is an adjustable, thin film, size and shape compliant, heating band and temperature sensor assembly attached to a linear translator. This device attaches to the objective and plugs into a separate Temperature Controller which regulates the objective temperature within 0.1°C of SETPOINT.

©12/23/98