Delta T4 Culture Dish System Instructions
Caution: Read Carefully Before Use!
The System is comprised of three main
components: a temperature controller, a stage adapter, and the Delta T
Dish. It utilizes a new and
improved technology that is much faster and more accurate than
traditional peripheral stage warmers.
The controller has a fast learning curve to anticipate changes in
the dish temperature due to perfusion or surface evaporation while
regulating temperature and compensating for entropy.
The fast response capability of this system enables control
features previously not possible with conventional peripheral systems.
Therefore, it is highly recommended that you read and fully
understand these instructions before use.
Before You Begin:
Check to make sure there is no visible damage in shipment and save
all packing materials.
The Controller box should contain; controller, power cord, 2.5mm
screwdriver, 6’ BNC cable, 1/8” jack, and 3 pin mini DIN connector.
Check the packing list and call your supplier if there are any
Control Interface Descriptions
be illuminated when the controller is turned on.
If it does not light, check the main power source, then the fuse
block located in the power input module where the AC power cord plugs
is the black button adjacent to the display.
It selects the source of the information displayed. When the controller is turned on, the default condition is to
display the setpoint value indicated by the light adjacent to the word Setpoint. The button selects one of a sequence of display options.
This button can be pressed at any time and has no effect on the
temperature regulation functions of the controller.
indicate the source of the value displayed on the numeric display.
is adjusted by a potentiometer accessible with the 2.5mm screwdriver
provided. To adjust the
setpoint make sure the numeric display is set to Setpoint,
then adjust the setpoint to the desired value.
See adjusting setpoint value.
If you need to change the setpoint value during your experiment
on a routine basis there is a remote setpoint jack on the back of the
controller. See Remote
the location where the calibration of the dish thermistor reading
circuit can be adjusted.
the location where the calibration of the thermistor reading circuit can
is the location where the non-dynamic voltage adjustment is made. This is the energy value sent to the dish when the controller
is in Imaging Mode. Adjust Volts must be selected for the display and the Mode
switch must be pressed for this value to appear.
The display will show a number that is 10 times the actual
voltage value being sent to the dish.
Adjustments can be made to this value by turning the small screw
located in the hole corresponding to Imaging Volts.
button changes the mode of operation of the controller from dynamic
feedback control to a fixed voltage set by the Adjust
Volts setting. When the
controller is in Dynamic mode, the amber LED will be lit next to the
mode switch. When the
controller is in Imaging mode, the green LED will light.
If the controller is in Imaging mode as a result of pressing the
mode switch, only the green LED will light.
If the controller is in Dynamic mode and a TTL voltage is applied
to the BNC jack on the back of the controller, both the amber and the
green LEDs will light. Caution!
Make sure that the controller is not left in the Imaging mode for longer
than 30 seconds. This will
not damage the controller but if the temperature in the dish deviates
more than the error margin during this time, the alarm will sound and
the controller will ignore all the safety interrupts.
The controller will continue to supply the constant voltage to
the dish until the controller is returned to the Dynamic mode.
(Red Button) resets the controller to the cold start condition and
enters the energy value used in Adjust Volts as a starting regulation
It is to be used when any of the following conditions occur:
substitutes precision resistors into the circuit that reads the
thermistors so that calibration adjustments can be made to a known
is a six pin mini DIN connector where the stage adapter is plugged in.
Panel of the Delta T4 Controller
Power line input, power switch, voltage selector, and fuses are all
incorporated in the AC Power Input Module.
The controller can operate between 90 - 260V 50-60Hz, or a 24V
battery can be used as an alternate power supply.
The fuse carrier contains two 0.5A slow blow fuses.
The fuses can be accessed by prying the fuse block out of its
socket with a small screwdriver wedged under the small slot in the
opening of the AC line receptacle.
When replacing the fuse block, make sure to align the arrow on
the outside of the fuse block with the closest voltage value to your
supply line voltage:
For 90-130 supply line voltage place the fuse block with the arrow
pointing to 110-120V.
For 180-260 supply line voltage place the fuse block with the arrow
pointing to 220-240V.
Lid Adjustment is the adjustment for the voltage sent to the Heated
Lid. Its range is from 0 to 5 V DC.
The nominal voltage is 2.5 V.
This voltage should be measured at the Heated Lid with an external meter.
It should be adjusted to a value just above the condensation
is the location to plug in the Bioptechs Heated Lid.
is a voltage output direct from two instrument amplifiers reading the
thermistors from channel A and B.
The output from this socket is temperature /10.
37.0°C = 3.70 Volt DC. Left pin is channel A, center pin is
channel B, and right pin is ground.
an analog input that allows the user to set the controllers setpoint
from an external DC source. The sleeve is earth ground.
A DC voltage can be applied to the tip of this jack equal to
temperature /10. i.e.: 28.5°C
= 2.85 Volt. When the jack
is inserted into the controller it will automatically switch to the
jacks value. When removed it will default to the setting on the front of
the controller. Use for
programming or cycling operating temperatures.
allows the user to adjust the voltage sent to the dish when the Fixing Switch is depressed. Range
from operating temp to 97°C.
is provided so that cells can be fixed on the stage of the microscope.
It can also be used for heat shock experiments.
The switch is recessed so that it is not actuated by mistake.
Caution! This switch should be used when you intentionally want to
overheat your cells. It
provides a nearly instantaneous heat shock to the dish.
is a BNC connection used to change the mode of operation of the
controller from a remote source such as a computer or slave from a
shutter trigger. The
default condition of the controller is dynamic control.
To change to Imaging mode, apply a TTL logic high and hold to
this BNC. The controller will remain in Imaging mode as long as the TTL
condition remains high as indicated by the LED on the front panel.
Delta T4 Controller Operation
Without any peripherals plugged into the controller, turn the unit
on using the main on-off switch located on the rear panel adjacent to
the power cord.
Set the display to "DISH".
The temperature display will read a number between -3.7 and -4.2.
Press and hold 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 the
Set the display to "SETPOINT".
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.
Plug in the stage adapter without a dish.
With the display set to DISH
it will read room temperature.
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 press the RESET
button immediately before placing a dish into the stage adapter.
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.
If a dish containing cells is still warm from prior incubation,
the Delta T4 will reestablish temperature in about 15 seconds.
If the contents of the dish are at room temperature, the Delta T4
will require »2 minutes to reach setpoint.
The dish should contain approximately 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.
Adjust the SETPOINT by
immersing the reference thermistor into the dish and read its
temperature by setting the display to "REFERENCE".
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 if
you are working at 37°C;
place a Bioptechs liquid crystal temperature indicator dot into the dish
and adjust the SETPOINT until it turns green.
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 you forget to turn off the controller, you can always press
the RESET button to reset the learning curve.
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 RESET
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 RESET button at
any time to reset the controller’s learning curve.
Use of the Delta T4 in Narrow Depth of Field
Dynamic control characteristics of the Delta T4 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 narrow depth of field imaging.
The controller is equipped with a circuit that bypasses the
dynamic temperature control mode and permits you to temporarily send a
constant DC current to the dish to eliminate the possibility of an
abrupt change of energy occurring as might happen in dynamic mode.
This prevents Z-axis displacement of the specimen during imaging.
The controller can be switched from its
default Dynamic control mode
to Imaging mode by several
Pressing the mode button on the front of the controller
Pressing the optional footswitch
TTL communication from an external trigger such as a acquisition
host or slave from a TTL shutter trigger
the Delta T4 in the auto (dynamic) mode when you are not scanning or
When you intend to acquire an image
activate the Imaging mode.
The value of the DC current can be adjusted with the
potentiometer labeled Adjust Volts.
A good starting point is 2.7V shown as 27.0 on the display.
After adjusting the Adjust Volts setting, return the display to the Dish
position and check to see that the value you have entered is adequate to
maintain the setpoint value for at least 15 seconds.
Make sure the controller is returned to the auto mode
immediately after the image is acquired.
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, make sure the two thumbscrews are loosened two turns. Place the dish into the stage adapter by matching the tabs on the dish with the offset openings in the stage adapter. Then, rotate the dish 15° CW until it comes to rest under the opposing metal spring tabs. Tighten the thumb screws* finger tight to secure the dish from moving. Dishes are removed by loosening the thumbscrews then rotating the dish 15° CCW prior to lifting the dish out of the retainer.
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 to the "REFERENCE" position.
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 the stage adapters is preventable and not covered under warranty.
Care and Cleaning:
insert and controller can be cleaned by hand wiping with mild soap and
water or alcohol but do not immerse or autoclave.
The stage insert should remain dry at all times.
If liquids are spilled on or into the stage adapter they must be
cleaned and the stage adapter dried immediately!
Delta T Dishes
Delta T 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 and 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
The thickness of the dish can also be
determined from the stock number on the dish package.
coverslip thickness / black
coverslip thickness / clear
*Custom configurations available upon
High Numeric Aperture Use:
If you are using high numeric aperture
(immersion) objectives 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,
glycerin 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
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.
Delta T Perfusion with the Delta T Micro-Perfusion Pump:
The Delta T Micro-Perfusion Pump is shipped with dual
channel tubing. When the
tubing pair is stretched around the rollers on the pump, the tube with
the black band will have a slightly smaller ID yielding a slower fluid
transport capability. Attach
the supply line from your media to the barb on the input side of the
black band pump tubing and another tube from the output side of this
tube to the selected irrigation port of the dish.
Attach tubing from the aspiration port of 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
the pump speed as needed for suitable perfusion rate.
©Bioptechs, Inc. 4/19/2001