1) A digital microfluidic (DμF) platform was developed to automate the liquid handling steps for differentiating embryonic stem cells into cardiomyocytes and assaying the resulting tissue.
2) The DμF platform improved upon existing manual methods by automating embryoid body growth, differentiation medium exchanges, and non-invasive assays of beating cardiomyocyte tissue using electric fields.
3) Beating cardiomyocyte embryoid bodies were cultured on the DμF platform for up to 8 weeks and responses to known chronotropic agents were measured non-invasively through impedance changes, demonstrating the potential of the system for screening cardiotoxicity.
A presentation on paper published in Biomaterials, this study evidences using plant material for developing vascular system and applications in organ regeneration with 3D printing
August 2012
You will hear about a patented LLNL optical diagnostic microscope design that can provide real-time imaging for tissue pathology and many other market applications.
ABSTRACT- The present study was conducted to investigate the effect of cadmium chloride on Histoarchiteceture of head kidney of fresh water fish Heteropneustes fossilis. The fishes were exposed to 0.5 ppm of cadmium chloride for 21 days. The most remarkable changes in head kidney, due to cadmium chloride were lysed condition of interrenal and chromaffin cells. The traces of cytoplasm had dark brown to black coloured cytoplasm. Most of cells are deformed and necrotic condition. Their size was significant at (P< 0.01 and 0.001) increased after cadmium chloride. All these changes will be recovered by herbal compound i.e. Ashwagandha. The damaged tissues were recovered in already treated group.
Key-words- Ashwagandha, Cadmium chloride, Chromaffin cells, Heteropneustes fossilis, Histopathology, Interrenal cells
A presentation on paper published in Biomaterials, this study evidences using plant material for developing vascular system and applications in organ regeneration with 3D printing
August 2012
You will hear about a patented LLNL optical diagnostic microscope design that can provide real-time imaging for tissue pathology and many other market applications.
ABSTRACT- The present study was conducted to investigate the effect of cadmium chloride on Histoarchiteceture of head kidney of fresh water fish Heteropneustes fossilis. The fishes were exposed to 0.5 ppm of cadmium chloride for 21 days. The most remarkable changes in head kidney, due to cadmium chloride were lysed condition of interrenal and chromaffin cells. The traces of cytoplasm had dark brown to black coloured cytoplasm. Most of cells are deformed and necrotic condition. Their size was significant at (P< 0.01 and 0.001) increased after cadmium chloride. All these changes will be recovered by herbal compound i.e. Ashwagandha. The damaged tissues were recovered in already treated group.
Key-words- Ashwagandha, Cadmium chloride, Chromaffin cells, Heteropneustes fossilis, Histopathology, Interrenal cells
SeedEZ 3D cell culture application notes - gel and drug embeddingLena Biosciences
SeedEZ 3D cell culture application notes - gel and drug embedding. Many inert polymers used as scaffolds for 3D cell cultures and colony formation are also used in drug delivery systems both in vitro and in vivo. Read this practical guide to learn how SeedEZ lets you merge these two worlds in order to integrate 3D cell cultures into standard drug delivery and testing applications.
By incorporating or adding drugs to SeedEZ, or in polymer matrices embedded in SeedEZ, dosage forms which release a drug over a period of time may be prepared in a desired shape and size. More importantly, all SeedEZ-based dosage forms may be tested in situ, with cells in a 3D cell culture. SeedEZ wicks most sol-state hydrogels, hydrogel precursors, semisolid media, excipient formulations, pharmaceuticals and test compounds. As a result, SeedEZ offers a novel 3D framework for (A) development of sustained release drug delivery systems that are simple to make and convenient to use in vitro; (B) localized or distributed drug delivery into 3D cell cultures using spot-a-culture and spot-a-drug approach, wick, dip or SeedEZ-stack method; (C) gradient formation and testing of drug combination strategies; (D) quality control testing and assurance; and (E) development of test platforms for quasi-steady drug release.
Notably, in most diffusion-driven drug delivery systems, a drug release rate declines in time. A degradable polymer matrix embedded in SeedEZ may enable quasi-steady drug release from a defined volume, defined by SeedEZ, when the matrix degradation rate is adjusted to compensate for this decline via increased drug permeability from the SeedEZ/polymer matrix system.
The application note covers use of common biomaterials, including extracellular matrix hydrogels (Collagen and Matrigel), gels from natural sources for spheroid cultures and controlled drug release (Agarose, Alginate, Methylcellulose, Gelatin), and synthetic materials such as Poloxamers (Pluronic - used for cell encapsulation, drug delivery and pharmaceutical formulations), and Carbomers used in ocular, transdermal, oral and nasal delivery systems.
Doris Taylor Building New Hearts: Regenerative Medicine Becomes a RealityKim Solez ,
Dr. Doris Taylor presents "Building New Hearts: Regenerative Medicine Becomes a Reality" at the Banff Transplant Pathology meeting in Vancouver October 5, 2015.
3D-Bioprinting coming of age-from cells to organsDaniel Thomas
Over the past decade, annual spending on pharmaceutical development to treat many endocrinological systems has increased exponentially.
Currently, preclinical studies to test the safety and efficiency of new drugs, use laboratory animals and traditional 2D cell culture models. Neither of these methods are completely accurate reflections of how a drug will react in a human patient.
A solution has emerged in the form of 3D-Bioprinting technology, developed for the scalable, accurate and repeatable deposition of biologically active materials. With advances in this biomanufacturing technology, durable biological tissues for use in testing new pharmaceutical products are now being harnessed and refined.
SeedEZ 3D cell culture methods and protocols - cell seedingLena Biosciences
SeedEZ 3D cell culture methods and protocols – cell seeding. User guidelines and protocols for cell seeding into the SeedEZ three-dimensional (3D) cell culture system. Learn how to make consistent 3D cell culture and co-culture models with the SeedEZ substrate using spot-a-culture, dip-and-culture and stack-and-culture methods. Learn how to use SeedEZ for contact co-culture and non-contact co-culture. Next, learn how to make 3D sandwich cultures and 3D tissue reconstructions modeling multi-layered tissues such as cerebral cortex in a SeedEZ stack. Detailed protocols for generation of brain 3D co-cultures using primary cortical neurons and mixed population of glia seeded with and without Matrigel extracellular matrix into the SeedEZ are provided.
To evaluate the Interaction of Mn(II), Fe(II), Co(II), Ni(II),Cu(II), Zn(II) And Cd(II) Mixed- Ligand Complexes of
cephalexin mono hydrate (antibiotics) And Furan-2-Carboxylic Acid To The Different DNA Sources. All the metal
complexes were observed to cleave the DNA. A difference in the bands of complexes .The cleavage efficiency of the
complexes compared with that of the control is due to their efficient DNA-binding ability and the other factors like
solubility and bond length between the metal and ligand may also increase the DNA-binding ability. The ligands
(Cephalexin mono hydrate (antibiotics) and Furan-2-Carboxylic acid and there newly synthesized metal complexes
shows good antimicrobial activities and Binding DNA , thus, can be used as a new drug of choice in the field of
pharmacy. And for formulating novel medicinal agents.
Multiorgan Microdevices for ADME Evaluatio and Drug Design:-
Multi-organ micro-devices are microfluidic devices that gives the information of human metabolism by connecting the fluidic streams from several on-chip in vitro tissue cultures with each other in a physiologically relevant manner. Multi-organ micro-devices can predict tissue-tissue interactions that occur as a result of metabolite travel from one tissue to other tissues in vitro. These systems are capable of simulating human metabolism, including the conversion of a pro-drug to its effective metabolite as well as its subsequent active metabolite and toxic side effects. Since tissue-tissue interactions in the human body can play a significant role in determining the success of new pharmaceuticals, the development and use of multi-organ micro-devices present an opportunity to improve the drug development process. The devices have the potential to predict potential toxic side effects with higher accuracy before a drug enters the expensive and time consuming phase of clinical trials. Further, when operated with human biopsy samples, the devices could be a way for the development of individualized medicine. Since single organ devices are testing platforms for tissues that can later be combined with other tissues within multi-organ devices, we will also mention single organ devices where appropriate in the discussion those seems large area of interest in current research for individualized medicine and drug resistance study.
SeedEZ 3D cell culture application notes - gel and drug embeddingLena Biosciences
SeedEZ 3D cell culture application notes - gel and drug embedding. Many inert polymers used as scaffolds for 3D cell cultures and colony formation are also used in drug delivery systems both in vitro and in vivo. Read this practical guide to learn how SeedEZ lets you merge these two worlds in order to integrate 3D cell cultures into standard drug delivery and testing applications.
By incorporating or adding drugs to SeedEZ, or in polymer matrices embedded in SeedEZ, dosage forms which release a drug over a period of time may be prepared in a desired shape and size. More importantly, all SeedEZ-based dosage forms may be tested in situ, with cells in a 3D cell culture. SeedEZ wicks most sol-state hydrogels, hydrogel precursors, semisolid media, excipient formulations, pharmaceuticals and test compounds. As a result, SeedEZ offers a novel 3D framework for (A) development of sustained release drug delivery systems that are simple to make and convenient to use in vitro; (B) localized or distributed drug delivery into 3D cell cultures using spot-a-culture and spot-a-drug approach, wick, dip or SeedEZ-stack method; (C) gradient formation and testing of drug combination strategies; (D) quality control testing and assurance; and (E) development of test platforms for quasi-steady drug release.
Notably, in most diffusion-driven drug delivery systems, a drug release rate declines in time. A degradable polymer matrix embedded in SeedEZ may enable quasi-steady drug release from a defined volume, defined by SeedEZ, when the matrix degradation rate is adjusted to compensate for this decline via increased drug permeability from the SeedEZ/polymer matrix system.
The application note covers use of common biomaterials, including extracellular matrix hydrogels (Collagen and Matrigel), gels from natural sources for spheroid cultures and controlled drug release (Agarose, Alginate, Methylcellulose, Gelatin), and synthetic materials such as Poloxamers (Pluronic - used for cell encapsulation, drug delivery and pharmaceutical formulations), and Carbomers used in ocular, transdermal, oral and nasal delivery systems.
Doris Taylor Building New Hearts: Regenerative Medicine Becomes a RealityKim Solez ,
Dr. Doris Taylor presents "Building New Hearts: Regenerative Medicine Becomes a Reality" at the Banff Transplant Pathology meeting in Vancouver October 5, 2015.
3D-Bioprinting coming of age-from cells to organsDaniel Thomas
Over the past decade, annual spending on pharmaceutical development to treat many endocrinological systems has increased exponentially.
Currently, preclinical studies to test the safety and efficiency of new drugs, use laboratory animals and traditional 2D cell culture models. Neither of these methods are completely accurate reflections of how a drug will react in a human patient.
A solution has emerged in the form of 3D-Bioprinting technology, developed for the scalable, accurate and repeatable deposition of biologically active materials. With advances in this biomanufacturing technology, durable biological tissues for use in testing new pharmaceutical products are now being harnessed and refined.
SeedEZ 3D cell culture methods and protocols - cell seedingLena Biosciences
SeedEZ 3D cell culture methods and protocols – cell seeding. User guidelines and protocols for cell seeding into the SeedEZ three-dimensional (3D) cell culture system. Learn how to make consistent 3D cell culture and co-culture models with the SeedEZ substrate using spot-a-culture, dip-and-culture and stack-and-culture methods. Learn how to use SeedEZ for contact co-culture and non-contact co-culture. Next, learn how to make 3D sandwich cultures and 3D tissue reconstructions modeling multi-layered tissues such as cerebral cortex in a SeedEZ stack. Detailed protocols for generation of brain 3D co-cultures using primary cortical neurons and mixed population of glia seeded with and without Matrigel extracellular matrix into the SeedEZ are provided.
To evaluate the Interaction of Mn(II), Fe(II), Co(II), Ni(II),Cu(II), Zn(II) And Cd(II) Mixed- Ligand Complexes of
cephalexin mono hydrate (antibiotics) And Furan-2-Carboxylic Acid To The Different DNA Sources. All the metal
complexes were observed to cleave the DNA. A difference in the bands of complexes .The cleavage efficiency of the
complexes compared with that of the control is due to their efficient DNA-binding ability and the other factors like
solubility and bond length between the metal and ligand may also increase the DNA-binding ability. The ligands
(Cephalexin mono hydrate (antibiotics) and Furan-2-Carboxylic acid and there newly synthesized metal complexes
shows good antimicrobial activities and Binding DNA , thus, can be used as a new drug of choice in the field of
pharmacy. And for formulating novel medicinal agents.
Multiorgan Microdevices for ADME Evaluatio and Drug Design:-
Multi-organ micro-devices are microfluidic devices that gives the information of human metabolism by connecting the fluidic streams from several on-chip in vitro tissue cultures with each other in a physiologically relevant manner. Multi-organ micro-devices can predict tissue-tissue interactions that occur as a result of metabolite travel from one tissue to other tissues in vitro. These systems are capable of simulating human metabolism, including the conversion of a pro-drug to its effective metabolite as well as its subsequent active metabolite and toxic side effects. Since tissue-tissue interactions in the human body can play a significant role in determining the success of new pharmaceuticals, the development and use of multi-organ micro-devices present an opportunity to improve the drug development process. The devices have the potential to predict potential toxic side effects with higher accuracy before a drug enters the expensive and time consuming phase of clinical trials. Further, when operated with human biopsy samples, the devices could be a way for the development of individualized medicine. Since single organ devices are testing platforms for tissues that can later be combined with other tissues within multi-organ devices, we will also mention single organ devices where appropriate in the discussion those seems large area of interest in current research for individualized medicine and drug resistance study.
Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. Mammalian cell culture technology has become a major field in modern biotechnology; mammalian cell culture refers to the cells of a mammalian, isolated from specific tissues (i.e. skin, liver, glands, etc.) and further cultivated and reproduced in an artificial medium. Cell culture technology is currently playing a major role in toxicity testing, cancer research, virology, genetic engineering, and gene therapy.
OBJECTIVE:
To observe the transfection of CHO and HEK cells with GFP
To observe the recombinant GFP using Western Blotting
To purify the transfected HEK and CHO cells using AKTA Pure Purification
Scalability of Cell Culture Processes in Single-use Bioreactors using Differe...KBI Biopharma
Niket Bubna, Cameron T. Phillips, Sigma S. Mostafa and AbhinavA. Shukla. KBI Biopharma, Durham, NC
253rd ACS National Meeting & Exposition
April 2-6, 2017 • San Francisco, CA
#acsSanFran • www.acs.org/SanFran2017
Launching digital biology, 12 May 2015, Bremenbioflux
Intro. It is not a secret that in biology laboratories hours of manual work are considered a compulsory part of the experiment. During a day of work, lab researchers have to pipette the right amounts of fluids in tubes, carry them from one machine to another, program and handle each machine individually, label and document carefully each step and then convert the results to data and analyse it. For a simple routine experiment, each of the mentioned tasks is performed at least 10 times/day. Past decade, a big effort has been done to produce machines (e.g., pipetting robots) that would automate some of the tasks in the lab. However, these machines were developed under the industrial mindset to maximize the throughput of a single task. Thus, these machines are of large size, task-specific, difficult to use (they usually come with dedicated drivers and software) and most importantly, extremely expensive. A solution is the use of digital microfluidics to enable the advance from automated biology to digital biology. In my vision, a digital lab should be:
• fully integrated, running all the tasks on the same machine
• easy to use, with a web-based software for biological design of new experiments and hardware control
• general-purpose, allowing easy reconfiguration and design of new experiments
• cheap, offering open-source and do-it-yourself assembly kits
Talk. In the talk, I will present an overview of the road to digital biology, covering all the main aspects, from computer-aided design to hardware production and biological applications.
Hands on. Also, prepare for some real engineering action :). I will execute live a biochemical application (enzymatic reaction of β-galactosidase with Xgal) on my homemade digital biochip. We will then discuss the current challenges in the development process and everyone will get a chance to play with the device. And of course, I will happily consider any engineering advice or idea you have :).
Tissue engineering in heart and valve failure management.
Brian Bender BE Symposium Poster
1. Lab-on-a-Chip Platform for Culturing and Assaying Cardiomyocyte
Tissue Derived from Embryonic Stem Cells
Contact Pads
Driving
Electrodes
Brian F. Bender† and Robin L. Garrell*
Department of Bioengineering, University of California, Los Angeles
Los Angeles, CA USA 90095
† bfbender@ucla.edu
Digital Microfluidics
Tissue Growth Long-Term Culture Conditions
Acknowledgments
1G. Vunjak-Novakovic, et al., Tissue Eng, 2010, 16,
169-187.
2J.A. DiMasi, et al., J Health Econ, 2003, 22, 151-185.
3A. Aijian, et al., JALA, 2014, 20, 283-295.
4R. Fobel, et al., Applied Physical Letters, 2013, 102,
193513.
5J.L. Perez-Diaz, et al., J Colloid Interf Sci, 2012, 1, 180-
182.
Glass
PDMS
ITO Parylene-C
Cytop™ Aqueous Buffer Cell Media
Spacer
Side view schematic of DµF device for EB culture.3 Through-holes, or ‘wells,’ have been fabricated into
the middle plate of the DµF device. Droplets of cell-suspension are dispensed and delivered to the
wells (100-120 V at 18.5 kHz) and are pulled into the well via capillary forces. Cells aggregate and
compact into EBs after ~24 h in culture. A modified fabrication process allows multiple stacked layers.
Digital (droplet) microfluidics (DμF) is a liquid handling platform that enables the
manipulation (dispensing, translating, splitting, and mixing) of discrete pico-microliter
sized droplets of liquid on a planar array of photolithographically patterned electrodes
through the controlled application of electric fields.
OffOn
Electrowetting:
Conductive liquids respond
directly to the electric field
(applied voltage)
OffOn
Dielectrophoresis:
Non-conductive (dielectric,
insulating) liquids respond to
electric field gradient
This work was supported by UCLA research funds and a Dissertation Year
Fellowship to B.F.B. Confocal laser scanning microscopy was performed at
the UCLA CNSI Advanced Light Microscopy/Spectroscopy Shared Resource
Facility, which is supported by an NIH-NCRR shared resources grant (CJX1-
443835-WS-29646) and an NSF Major Research Instrumentation grant
(CHE-0722519). Cleanroom fabrication was performed at the UCLA CNSI
Integrated Systems & Nanofabrication Cleanroom (ISNC). Human
embryonic stem cells were graciously donated from the Nakano lab at
UCLA. Parts, training, and assistance with the DropBot System and µDrop
software were provided by the Wheeler lab at the University of Toronto.
References
Medium exchange from hanging drops.3 Medium
exchange was performed after 48 h periods with
differentiation medium.
Live/Dead Staining. EBs of
human fibroblasts (a) and
mouse mesenchymal stem cells
(b) maintained over 95% viability
after 48 h of incubation. A hESC-
EB (c) in an on-chip well. White
scale bar = 1 mm.
Embryoid
Body
Embryoid
Body
Differentiation
medium 2
1
3
Discarded
4
Liquid exchange. >50% exchange can be
achieved after two cycles, which is
sufficient for EB culture.
50
75
100
36.5
36.75
37
1 10 100 1000
RelativeHumidity(%)
Temperature(⁰C)
Time (log[min])
0
0.005
0.01
0 100 200 300 400 500
InstantaneousVelocity
(mm/s)
Time (ms)
60% RH
70% RH
80% RH
90% RH
0
0.005
0.01
0 100 200 300 400
InstantaneousVelocty
(mm/s)
Time (ms)
60% RH
90% RH
Velocity of DI water actuated at 80 V and 20
kHz from within an incubator, highlighting
that as the humidity rises the droplet
velocity decreases. The error bars correspond to
the standard error with n=6.
0
1
2
BeatingFrequency(Hz)
5 mM Epinephrine
5 µM Epinephrine
5 mM Caffeine
5 µM Caffeine
Media
A vertically stacked DµF design enabled EB
retrieval for downstream processing. Scale bar =
1 mm.
Capacitance and impedance measurements of an 8-day old beating
cardiomyocyte EB showing a more homogenous yet weaker beating profile
compared to the more mature but heterogeneous 8-week old beating
cardiomyocyte EBs.
A digital microfluidic (DµF) method for automating the liquid handling steps for
differentiating and assaying embryonic stem cell (ESC)-derived cardiomyocytes
was developed. This method improves upon existing methods by automating
the manual, multi-step, liquid handling protocols currently used for embryoid
body (EB) growth and differentiation. The electric fields used to manipulate
discrete droplets in DµF platforms were then used to non-invasively assay
phenotypic behavior in beating, 3D cardiomyocyte tissue.
1
Two Primary Motivations
Non-Invasive Assays
The stability over temperature,
humidity, and CO2 provided by
operation within an incubator helps
prevent evaporation, better maintain
solution concentrations, and mitigate
temperature shock.
Setup
Evaporation
Materials
Droplet Velocity
The material selection needed to be re-evaluated to avoid dielectric
breakdown and delamination.
Parylene-C is a CVD-deposited polymer commonly used as the dielectric
layer. After 24 hr of incubation, electrode actuation caused delamination
and device breakdown, revealing the need to readdress material selection.
SiO2 delaminated from ITO electrodes occurs without adherence to
strict cleanroom protocols. However, proper protocols produced
devices that could withstand incubated conditions for >1 month.
DµF operation from within an incubator has many advantages over the current
practice of moving a chip back and forth between the incubator and the benchtop.
Automating droplet sequencing requires a
precise understanding of droplet position and
translation speed for timing.
𝐹 𝑉 = 𝛾 𝐿𝐹 cos 𝜃 𝑎 𝑉
The surface tension of the water-air
interface, 𝛾 𝐿𝐹, decreases as the humidity and
the temperature rise.5 The actuation force,
𝐹 𝑉 , therefore decreases in the incubator.
Furthermore, the static contact angle, cos 𝜃 𝑜,
will decrease an the elevated humidity.
∆𝑃 =
𝛾 𝐿𝐹
𝑑
cos 𝜃 𝑎 − cos 𝜃 𝑜
𝐹(𝑉) = )𝑃𝑟 − 𝑃𝑎(𝑉 ∙ ℎ
The actuation force is therefore decreased
when modeling the difference in Laplace
pressure, ∆𝑃.
In a common, water-jacked incubator
the equilibrium conditions can take
hours to reach.
The concentration of solution constituents can change dramatically
even in samples placed into an incubator that must re-equilibrate.
This can alter cell behavior and confound results.
Differentiation Medium Day 3-5 Day 5-7 Day 7-10
Constituent Stock Final Final
No Chemical
KY-1 50 mM 3 µM 3 µM
XAV 10 mM 1 µM 1 µM
A419259 5 mM 0.3 µM 0.3 µM
AG1478 100 mM 8 µM 4 µM
Day 3 – 10: 0.4 % Albumin Media
Day 10+: 0.04 % Albumin Media
(C)
Known chronotropic and ionotropic agents were
delivered on-chip to beating cardiomyocyte EBs.
Video recordings revealed expected results.
Our understanding of cardiomyocyte maturation is not fully developed.1
Many methods have been developed to probe the differentiation and maturation
process of cardiomyocytes, such as molecular or genomic tagging, videographic
algorithms, micro-post arrays, and micro-electrode arrays. However, few methods have
been developed to holistically monitor 3D tissue samples, despite our awareness that
these systems can illicit more in-vivo-like behavior.
2 A need exists for streamlined tools for differentiating and assaying cardiomyocytes.1
A recent study has estimated the cost of developing a single new drug at over $2.6 billion
and taking over 25 years.2 In order to bring down the costs and timelines for screening
and developing new therapeutics, advanced cell-based assays that better mimic in-vivo
conditions are needed. DμF is capable of automating the liquid handling and non-
invasive assaying steps needed for a streamlined stem cell culture microenvironment.
Abstract
Conclusions
1
A new approach was taken to electrically monitoring EB beating behavior. Changes to
beating heterogeneity were observed in whole 3D samples by applying an electric field
around the entire sample and monitoring impedance changes.2
A DµF platform was used to create a streamlined lab-on-a-chip device capable of
forming EBs of human embryonic stem cells, long-term culturing in an incubator,
differentiation into functional cardiomyocytes, and performing non-invasive assays.
Epinephrine produced a ~4-5x
increase in beating amplitude
evidenced via similarly shaped
capacitance measurements.
8-day old beating EB 8-week old beating EB Controls
19-point weighted triangular smoothing
applied
Epinephrine Assay
Electrical
Visual EB Retrieval
~1-3 mm
~7-10 μL
4 μm
400 nm
110 nm1.7 mm
1 mm
2 mm
To CPU
300 μm
Beating cardiomyocyte EBs were positioned between electrodes, and an 80 V,
15 kHz AC signal was applied while the impedance/capacitance was recorded.
Embryoid Body
(EB)
Differentiation
DropBot4 equipment
and amplifier Cell culture
incubator
3D-printed plastic
holder for both the
DµF chips and the
printed circuit
board cabling
connections