804A: Permeabilized Fiber ATPase Apparatus
The 804A ATPase apparatus was designed to enable physiology researchers to measure consumption of ATP along with contractile force and length changes of permeabilized fibers. The 804A works in conjunction with an Aurora Scientific model 315D or 322D high-speed length controller and an Aurora 400C series force transducer.
The 9-well bath plate is corrosion resistant and can be used for permeabilized fiber experiments such as force-pCa and kTr. Bath 1 is the largest bath which includes right angle prisms on either side, allowing simple fiber attachment and accurate measurement of cross-section. The first bath also has a glass bottom which allows the fiber to be imaged with an inverted microscope or to set the resting sarcomere length (SL) using our High-Speed Video Sarcomere (HVSL) system. A laser diffraction system is also included for setting SL of cardiac tissue. The 804A also includes an add-on ATPase cuvette that is used to measure ATP consumption. The ATPase cuvette includes optics to excite the fluorophores in solution with UV light (970A) and to quantify the absorbed light signal from the bath using a photodetector (971A).
Utilizing a microprocessor-based controller and our 600A software, the researcher can define which bath the fiber is placed in at specified time points, simplifying a force-pCa test. Position sensors ensure bath placement accuracy to better than +/- 0.05mm. A built-in syringe pump provides nL accuracy for ADP injection into the ATPase measurement cuvette. Each apparatus includes XYZ micrometer stages with built-in mounts for Aurora Scientific high-speed length controllers and force transducers.
Components
825A Thermocouple Meter / TEC Controller:
Thermometer and TEC Controller controls bath temperature from 4 to 38°C
826A Water Circulator:
Water circulator that provides heating/cooling for 804A apparatus. Includes water reservoir, pump, and radiator with fan
970A UV Light Source:
Dual-wavelength UV LED light source (340nm and 430nm) provides excitation for the fluorophores in the ATPase cuvette via an included fiber optic cable
971A ATPase Photodetector:
Dual-wavelength UV photodetector provides measurement of the intensity of the 340nm and 430nm wavelengths from the ATPase cuvette via an included fiber optic cable.
Stories of Success
804A – Permeabilized Fiber ATPase Apparatus
CHALLENGE
In 2016 Aurora Scientific was approached by Dr. Pieter de Tombe and Dr. Ramzi Khairallah to partner in the development and commercialization of a new apparatus for making measurements of ATP consumption in permeabilized tissue. This initiative was in response to a growing demand for these types of measurements after a long period of sparse publications on the subject. With no commercial ATPase systems available, and with few researchers able to construct their own systems, a need for a new product was apparent. This new product required motor-driven movement of the bath plate and sensitive, affordable, and easy-to-use, optics for measuring ATPase activity.
SOLUTION
After many prototypes and iterations, a version of the apparatus that could mount onto an inverted microscope was developed. Aurora Scientific was able to leverage their experience with precise motion control from the existing 802D fiber apparatus to develop a reliable and relatively inexpensive moving chamber. Together with Dr. de Tombe and Dr. Khairallah’s extensive knowledge of optics, this solution has opened the door to new technologies that will achieve much improved ATP resolution.
RESULTS
The 804A Permeabilized Fiber ATPase Apparatus represents a true collaborative effort between industry and academia in the spirit of scientific discovery. Aurora Scientific is proud of its development and is grateful for the efforts of all those involved.
Related Products
315D/322D: High-Speed Length Controllers
The 315D/322D High-Speed Length Controllers give physiologists the ability to control and measure length of single cells, fibers and whole muscle with ease.
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400C: Force Transducers
The 400C series of force transducers is the next generation of our widely used 400A and 400B series. The 400C series enables contractile measurements from a variety of muscle types and sizes and are designed to meet the needs of muscle researchers.
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600A: Real-Time Muscle Data Acquisition and Analysis System
The 600A Digital Controller serves to integrate components and provide the researcher control of system operations, data collection and signal analysis.
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825A: Thermocouple Meter / TEC Controller
Thermometer and TEC Controller controls bath temperature from 0 to 40°C
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901D: High-Speed Video Sarcomere Length
A simple, turn-key system for measuring sarcomere spacing in isolated muscle preparations.
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902A: Laser Diode Sarcomere Length
A simple, easy to use laser diode system designed for measuring sarcomere length in thicker muscle fibres
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1400A: Permeabilized Fiber System – Microscope Mountable
The 1400A and 1410A Permeabilized Fiber Systems are designed to enhance experimental throughput and simplify complex permeabilized fiber experiments.
Learn MoreSpecifications
| Bath Plate | |
|---|---|
| Number of Baths | 10 including: 1 Relax/Mounting, 8pCa, 1 ATPase |
| Bath Dimensions [mm] | Bath 1 – Relax: 16L x 6W x 3D Baths 2-9 – pCa: 16L x 3W x 3D Bath 10 – ATPase: 4L x 1W x 5.2D |
| Bath Positioning Accuracy [mm] | +/-0.05 |
| Bath Volumes [µL] | Bath 1 – Relax: 295 Baths 2-9 – pCa: 124 Bath 10 – ATPase: 21 |
| Inter-Bath Distance [mm] | Bath 1-2: 14 Baths 2-9: 7 Bath 10: 14 |
| Indexing Time [second] | Bath1–2: 1.3 Adjacent Baths 2-9: 1.1 Bath 9-10: 1.3 Bath 1-10: 2.7 |
| Z-Axis Excursion [mm] | High Sensor-Up Sensor: 5.75 Up Sensor-Down Sensor: 4.0 Down Sensor-Lowest Position: 3.75 Total Maximum Excursion: 13.5 |
| Right-Angle Prism Dimensions [mm] | 6L x 3D x 3H |
| Material | Apparatus: 6061-T6 Aluminum – Black Anodized Bath Plate: 6061-T6 Aluminum – Hard Anodized |
| ADP Injection Syringe | |
|---|---|
| Available Volumes [µL] | 10, 25, 50 |
| Type | Hamilton type RN1700, cemented needle |
| Syringe Dimensions [mm] | 1701 – 10µL: 0.46 ID x 60 Stroke 1702 – 25µL: 0.73 ID x 60 Stroke 1705 – 50µL: 1.03 ID x 60 Stroke |
| Flowrate Range [pL/sec] | Max: Min: |
| Pump Stroke [mm] | 52 |
| Minimum Injection Volume [pL] | 38 |
| Maximum Injection Volume [µL] | 1701 – 10µL: 8.7 1702 – 25µL: 21.7 1705 – 50µL: 43.3 |
| Laser SL Measurement | |
|---|---|
| LED Laser Power [mW] | 5 |
| Beam Dimensions [mm] | 3 x 1 |
| Wavelength [nm] | 635 |
| Laser Class – CDRH | IIIb |
| Measurement Range [µm] | 1.7 – 10.0 |
| Temperature Control | |
|---|---|
| Type | 2 x Peltier modules |
| Temperature Control Range [°C] | 4 – 40 |
| TEC Power [W] | 33 |
| Voltage [V] | 12 |
| Temperature Sensor | AD590 |
| General Specifications | |
|---|---|
| Operating Temperature [°C] | 10 – 30 |
| Power Required | 100, 120, 220, 240 VAC, 50/60 Hz. available |
| Power Consumption [W] | 20 |
| Weight [kg] | 2.5 |
| Dimensions [mm] | Microscope Mount Configuration: W x D x H***** Table Mount Configuration: W x D x H***** |