802D: Permeabilized Fiber Apparatus – Microscope Mountable
The Aurora Scientific 802D Permeabilized Fiber Test Apparatus is designed for ease-of-use, allowing researchers to automatically measure fiber dynamics at varying pCa with the push of a single button.
The eight well bath plate is corrosion resistant and allows the researcher to test multiple solutions without the need to empty and re-fill baths. Bath 1 is the largest bath to allow easy fiber attachment and includes right angle prisms on either side for accurate measurement of fiber cross section.
Each apparatus contains XYZ micrometer stages with built-in mounts for Aurora Scientific high-speed length controllers (315C, 322C) and force transducers (400A Series). Utilizing a microprocessor-based controller and our 600A software, the researcher can define which bath the fiber is placed in at specified time points.
Importantly, moving the bath plate rather than the fiber ensures a much more accurate measurement of force and reduces the chance of tissue damage. Position sensors ensure bath placement accuracy to better than +/- 0.025mm.
Components
825A Thermocouple Meter / TEC Controller:
Thermometer and TEC Controller controls bath temperature from 0 to 40°C
826A Water Circulator:
Water circulator that provides cooling for 801C, 802D, 803B apparatus. Includes water reservoir, pump and radiator with fan
Variants
Bath Size Variants:
802D-120 Permeabilized Fiber Test Apparatus, 120µL bath
802D-160 Permeabilized Fiber Test Apparatus, 160µL bath
Temperature Jump Variant:
802D-120-TJ Temperature Jump Permeabilized Fiber Test Apparatus, 120µL bath – (120µL bath with separate temperature control for baths 1 & 2 and for baths 3 through 8, includes 2 825A TEC Controllers)
Stories of Success
802D – Motorized Bath for Permeabilized Fibre Experiments
CHALLENGE
In 2003 Prof. John Faulkner’s Lab at the University of Michigan was about to undertake a multiyear test program of over a thousand permeabilized fiber samples from humans. They needed a method of automating calcium switching.
The sheer volume of fibers that needed testing required a system that was robust enough to be used by undergraduates on a daily basis, with no damage to force transducers or motors. No commercially available apparatus existed at the time and any home-made units suffered from reliability issues.
SOLUTION
After consultation with Faulkner’s lab, Aurora Scientific designed and built the 802A permeabilized fiber apparatus featuring 6 wells, temperature control and switching speeds of about 1 second. The chamber was integrated as a system with our force transducer and motor. The apparatus could repeatedly place the bath at the exact location every time, without damage to the fiber or test equipment.
RESULTS
Three complete systems were supplied to the Faulkner lab for the price of home-built units. These systems have been in almost continuous operation for the past 6 years and thousands of fibers have been tested by many different undergraduate workers. Updated versions of this system have been acquired by many of the leading fiber research labs worldwide.
Select References
- Larkin, Lisa M. et al “Weakness of whole muscles in mice deficient in Cu, Zn superoxide dismutase is not explained by defects at the level of the contractile apparatus.” Age (2013) DOI: 10.1007/s11357-012-9441-7
- van der Pijl et al. “Titin-based mechanosensing modulates muscle hypertrophy” Journal of Cachexia, Sarcopenia and Muscle (2018) DOI: 10.1002/jcsm.12319
- Fukutani et al. “Influence of residual force enhancement and elongation of attached cross-bridges on stretch-shortening cycle in skinned muscle fibers” Physiological Reports (2017) DOI: 10.14814/phy2.13477
- Zhang, Tan et al. “Human slow troponin T (TNNT1) pre-mRNA alternative splicing is an indicator of skeletal muscle response to resistance exercise in older adults” The Journals of Gerontology: Series A, Biological Sciences and Medical Sciences (2014) DOI: 10.1093/gerona/glt204
- Manders, Emmy et al. “Diaphragm weakness in pulmonary arterial hypertension: role of sarcomeric dysfunction.” American Journal of Physiology-Lung Cellular and Molecular Physiology (2012) DOI: 10.1152/ajplung.00135.2012
- Klaiman, Jordan M. “Cold acclimation increases cardiac myofilament function and ventricular pressure generation in trout.” The Journal of Experimental Biology (2014) DOI: 10.1242/jeb.109041
- Lee, Eun-Jeong, et al. “Calcium sensitivity and myofilament lattice structure in titin N2B KO mice.” Archives of Biochemistry and Biophysics (2013) DOI: 10.1016/j.abb.2012.12.004
- Gineste, Charlotte, et al. “Alterations at the Cross-Bridge Level Are Associated with a Paradoxical Gain of Muscle Function In Vivo in a Mouse Model of Nemaline Myopathy” PLoS ONE (2014) DOI: 10.1371/journal.pone.0109066
- Bezold, Kristina L. et al. “A gain-of-function mutation in the M-domain of cardiac myosin-binding protein-C increases binding to actin” Journal of Biological Chemistry (2013) DOI: 10.1074/jbc.M113.474346
- DeWinter, Josine M. et al. “Troponin activator augments muscle force in nemaline myopathy patients with nebulin mutations.” Journal of Medical Genetics (2013) DOI: 10.1136/jmedgenet-2012-101470
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Learn MoreSpecifications
| Bath | 802D-120 | 802D-160 |
|---|---|---|
| Number of Baths | 8 | |
| Bath Volume [µL] | Bath 1: 198 Baths 2-8: 120 |
Bath 1: 198 Baths 2-8: 158 |
| Bath Dimensions [mm] | Bath 1: 14 L x 5 W x 3 D Baths 2-8: 14 L x 3 W x 3 D |
Bath 1: 14 L x 5 W x 3 D Baths 2-8: 14 L x 4 W x 3 D |
| Right Angle Prisms [mm] | 3 W x 3 H x 6 L (located in Bath 1) | |
| Bath Angular Rotation, Z-axis [°] | 3.0 | |
| Bath Excursion, Z-axis [mm] | Maximum1: 4.72 Minimum2: 3.35 |
|
| Indexing Time [s] | Bath 1-2: 1.2 Adjacent Baths in range 2-8: 1.1 Bath 1-8: 2.5 |
1 = Force Transducer End of Bath; 2 = Length Controller End of Bath
| Apparatus | 802D-120 / 802D-160 |
|---|---|
| Number of Stepper Motors | 2 |
| Motor Voltage [V] | 12 |
| Stepper Movement [µm/step] | 25 |
| Thermoelectric Coolers | 802D-120 / 802D-160 |
|---|---|
| Number of TECs | 2 |
| Control Temperature [°C] | 0 – 40 |
| Power [W] | 33 |
| Voltage [V] | 12 |
| Thermocouple Microprobe | 802D-120 / 802D-160 |
|---|---|
| Model | Physitemp IT-18 |
| Type | T Thermocouple Copper-Constantan |
| Time Constant [s] | 0.1 |
| Construction | 0.064 mm diameter Teflon tube with sealed tip |
| General Specifications | 802D-120 / 802D-160 |
|---|---|
| Power Required | In: 100-240 VAC, 50/60 Hz., 1.8A Out: 12VDC @ 2A |
| Power Consumption [W] | 25 |
| Weight [kg] | 2.4 (with stand), 1.5 (without stand) |
| Dimensions3 [cm] | 18.1 W x 25 L x 6.9 H |
3 = Apparatus without stand