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I am going to do sequencing for the first time and I am a bit confused how to count amount of chips that I need. I will use SMARTer smRNA-Seq Kit Takara for 96 reactions for library preparation. Should I calculate amount of samples per chip according to the size of my fragments? Or according to the concentration of pooled samples after library preparation?
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Are you trying to use more than 1 flow cell and split your samples across different flow cells with different runs? You should try to avoid batching if you can do not deal with potential batch affects. You need to think about what does reads will do, are they for counting or genotyping. If you are counting, do you really need 1000 coverage per fragment, would 100x per fragment be OK for statistical purposes? Generally the more samples you multiplex the less reads/coverage per sample and vice versa. The NGS library kit should have some guidelines in the handbook for how many samples to multiplex given the size of the panel for a given panel size and coverage. Think of this, if you put 24 sample and got 30,000,000 reads per sample on average, then if you put twice as many samples it will roughly be 15 million reads per samples if you get similar cluster pass filter and total output as the run with 24 samples and 30 million reads (not everyone has the same total Gb output). I would highly recommend you contact a field application scientist or someone who has used to kit to engage you on this topic if the above mentioned guidelines aren't in the kit handbook.
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Considering any CMOS technology, I am eager to know what is the upper limitation of MOSCAP design. Also I want to know that MOSCAP is larger than conventional on chip capacitors or not. For instance how to compare 1pF MOSCAP and a 1pF on chip capacitor regarding die occupation. Totally avoiding passive elements in chips can save lots of area, so I am looking for methos for realizing larger capacitors with smaller die occupation.
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ESR (Equivalent Series Resistance) is a limiting factor for capacitors. A large value capacitor (tantalum, electrolytic, etc) has a high ESR. It will not filter high frequencies. A ceramic capacitor has a low ESR. It will filter high frequencies.
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What percentage of microbial biofilm on a MBBR Chip is required for a industrial wastewater treatment plant?
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Simply higher is better, colonization of media in MBBR system is important to ensure process effectivity and also to prevent microbes from wash out from the reactor.
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I want to make a micro-fluid chip using PDMS. I print the casing for this purpose using SLA material. I want to cure the chip faster. I have a microwave oven (700 Watt). My question is that can I use microwave for curing PDMS? And what will be the effect on SLA part?
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Ahmed Abrar if you can control the temperature of the microwave oven it will work better, like using PWM pulses. This may require some changes in normal household microwave oven. During my experiments with microwave oven, I noticed that if I keep heating the PDMS in the microwave oven for more than 1 mint, it was creating problem. You can test 30 secs heating, like heat and give it sometime to cool down and heat again.
For second part of your question, No I did not use any other type of oven.
Thanks
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Hi,
Has anyone doing microfluidics work for cell culture started having problems recently with getting their Sylgard 184 PDMS to reliably and permanently bond to the glass substrate? We have tried replacing our PDMS elastomer base and curing agent recently but this has not fixed our problem. We have also tried using 10% less curing agent which didn’t help either.
Briefly our protocol involves the initial bake at 60 degrees C for 1.5 hours, cutting out chips the next day then cleaning the PDMS surface with adhesive tape, cleaning the glass surface with 70% ethanol and a blast of nitrogen, plasma treatment for 30 seconds at 50% power/~15W (Zepto ONE, Diener Electronic plasma cleaner), and then sticking the surfaces together. Final annealing overnight at 60 degrees C before checking my chips for any bonding issues.
Normally when I have improperly bonded chips it’s immediately obvious and I’d see a portion of the bonded PDMS stuck onto the glass. Recently I’ve noticed that too many of the chips come off the glass cleanly, leaving no residual PDMS. I also sometimes find that a chip may be properly bonded but then it comes undone at a much later stage. All this leads me to think that the bonding is no longer permanent like it should be. I’d appreciate any feedback or advice, or if anyone else has noticed this and what may have worked for you if you did. Thanks very much in advance!
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Plasma treatment power is too lower than ours, sometimes humid environments OH- group easily & fastly disappears. I suggest using 100% power to treat. For reference, we use 70W O2 20sccm 30sec condition. Abigail Dos Santos
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I have been using the following video to make my connections, but I am stuck where the serial monitor is showing - "Connecting....." Can't figure where I am going wrong.
let me know as soon as possible
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WiFi with ESP8266 works well, and is a good start, even for microcontroller beginners. However, I am not sure if RG is the best place to ask such kind of technical questions. I would recommend to place your question on StackOverflow instead. Use the correct tag [esp8266], post your code sketch, and provide enough information about your setting.
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Hello,
I was wondering if someone can explain to me how to couple the light from a fiber into a waveguide on a chip.
Thank you in advance.
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Dear Dr Sewidan..I guess that you would like to couple from a conventional (monomode?) fiber to a dielectric ? waveguide on a chip (optical image line?)We may also discuss this via normal e-mail (Fritz Caspers@cern.ch
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Dear Colleagues, I am working with gas sensing nanomaterials now and frequently see that many authors utilize 6-pin chips with steel mesh cover to fabricate the prototype gas sensors. The chips resemble something like dummy MQ2, MQ5, or MQ-135 modules for Arduino. However, I could not find on the internet any suppliers of such dummy/empty chips. (It is easy to find numerous ready chips with an installed sensing layer though.) Could you, please, suggest some suppliers which can provide the dummy chips, better in EU.
Sincerely yours,
Alexander.
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Dear All
I would like to order a PDMS chip from a company by directly sending my AutoCAD dwg file and with explaning channel height-size specifications. Then, is it possible to receive a PDMS chip or at least a mask (so I will just need to pour PDMS and create chips)? if yes, do you recommend any companies? Many thanks!
Best Regards
Su
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Hi, for anyone still looking, Wunderlichips' core business is providing PDMS devices based on researchers' designs. https://wunderlichips.ch/en/home
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Hello everyone,
The workshop in my lab printed a mold with a Formlab resin printer (black resin) to make a simple microfluidic chip.
I'm aware that photoinitiators in 3D printer resins can inhibit the cure of PDMS.
I tried several cleaning/curing of the resin and everytime my PDMS (1:10 sylgard 184) is overall ok, except for the parts in contact with the mold which is milky and slightly sticky.
This surface results in no correct bonding with glass after plasma treatment...
So my strategy was to cast an intermediate polymer on the PDMS chip and then recast PDMS on top to get my final crystal clear PDMS working chip.
I tried with what we had in the lab > NOA73 that I pushed to 10min UV exposure for curing but surprise I have the exact same: a milky side (in contact with the milky side of the PDMS) and a clear side (air side).
When I cast PDMS on that it's curing everywhere except where the NOA is (fully unpolymerised).
I read that PDMS casting was ok on NOA, I even found a method paper doing this.
So I wanted to know if someone has struggle with this kind of materials before and if they found a solution. I precise that I need the flexibility of the PDMS for my final chip (embeded electronic valves and tubing).
I will certainly try epoxy and polyurethane as intermediates.
Thank you,
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Hi Nelson,
I had the same problem with Anycubic Photon printer's resins. In our lab, we use Mann Ease Release 200 nonadhesive spray. After spraying (just a thin layer), place your molds on the hot plate (2 hours, 80 degrees) to create a thin and uniform covering layer.
It works fine, but still be ready, that 1 or 2 PDMS platforms will be just for the cleaning step.
The PDMS-glass, PDMS-PDMS bonding works well.
Cheers,
Michael.
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Describe:As the picture shows,I want to study little chips during cutting process ,like circled by the red line.But what I did is the second figure ,there are big chips rolling,no little chips drop or crash fly.
My sets:
1.PARTS:I set the tool as a rigid body in the interaction step,the work piece is separated to 2 or 3 parts for a tighter mesh in separate part.
2.PROPERTY: As the figure 2 shows ,JC hardening , JC damage , Shear damage . I am not sure if my defination is accurate.
3.MESH:Just like the figure 4 shows.
That's my question above , can anybody give some tutorial or advice , I am very appreciate about your reply.
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Use finer mesh in contact region with element deletion criteria.
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I am using Keithley 2450 source meter to measure dark current of commercially available silicon PIN photodiode chip (SLCD - 61N - 5). As mentioned in the data sheet of the photodiode, the dark current measured at 5V reverse bias is around 3 microamperes. But, I am getting a dark current of around 20 milliamperes.
Connecting wires are attched to the anode and cathode of the photodiode chip using conductive silver paste. The Keithley 2450 source meter and SIlicon PIN solderable photodiode chip(SLCD-61N-5) are connected by using a breadboard. The measurements are taken using the front panel 4 probe method.
I am attching the datasheet of the photodiode chip and the circuit that I have used to measure the dark current.
I would be grateful if anybody could help me to sort out this problem.
Thank you for your valuable time.
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It may simply be that the component does not fully follow the standard. Often such components have a minimum value, maximum value and "typical value". The latter is often stated in the data sheet. Have you tried several components of the same type? Possibly checked if the measuring instrument is correct?
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I want to do Chip assay for detection of three different transcription factors forming a complex and bind to DNA at the same locus.
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You can prepare your sonicated, cross-linked chromatin and then divide into three aliquots (four including the IgG control). Incubate each with antibody of choice, pull down (lots of kits for this, Active Motif has a straightforward one) and then qPCR the locus of interest. If all three proteins are bound there, all three antibody pulldowns should give a qPCR result much higher than background (IgG). Separate assays would likely have to be used to validate that those proteins are part of the same complex. I'm not aware of the assay to simultaneously confirm the presence of proteins in a complex and pull down the entire complex to assess bound DNA loci. If it is being done it seems that it would be quite complex/finicky and dependent on each unique protein complex.
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I am looking to find a technique that will allow me to pull down a promoter with a oligo-pull down method and identify any proteins that are in contact with that promoter. I have never seen any such technique in a paper but it would be a very good way to answer the question I am hoping to ask. Is anyone familiar with a technique that can do this or something similar to this? Thank you!
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Hi, my friend try this method before, using ~500bp biotin-labelled DNA to pulldown interacted proteins from the cell lysate. You can try to look the method here:
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Hi there!
I have a question:
What is the best photonic network on chip simulator which has a good manuals and learning and community?
best
Yasin Asadi
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Hi Yasin
I think the best option is Lumerical Interconnect. In this module, you can either define your own device or just use the predefined elements like MZIs, ring resonators, modulators, and detectors to build your own optical network. It can integrate with KLayout for chip design and test, and also provides a reliable simulation results based on TMM or S-matrix method. Besides, the Lumerical "knowledge base" website is available for all users where you can either use it as a learning tool or debugging your simulation procedure and coding.
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Once the RNA is subjected to first-strand cDNA synthesis, is it stable? Can this product be run on the cDNA HS Chip on the bioanalyser?
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Shilpa Patil As of my experience, first-strand cDNA is more unstable than double-stranded DNA, therefore, more care is required during storage. But slightly stable.
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I have a stainless steel 2 mm thick raw material. A 3 diameter hole is to be punched into it. My punch is made of ASP2023 or HSS material. It is giving a life of only 3000 - 4000 strokes before chipping off.
Which is the best coating to be used to increase the life of my punch. The target punch life that I need is 70000.
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I would like to extract chromatin from frozen samples, but most of them are embedded in OCT?
Does anyone has experience with it?
Thanks a lot!
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Hélène Neyret-Kahn how did you remove the OCT from the frozen tissue to further extract chromatin. Did you use a specific kit?
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Hi,
I would like to know if any of you have sonicated the cell line K562 with Bioruptor for ChIP. How many cycles do we need to properly sonicate this cell line?
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Hi Lara,
I am performing CHIP on human AML cell line (MV4-11). I want to sonicate the samples for 30 sec ON and OFF. May I know how many cycles shoule be enough to fragment the chromatin ?
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At present, I am performing an isothermal amplification experiment on the PDMS chip (heating at 60°C for 60 min), but I have encountered a big problem. In many cases, the droplets in the PDMS cavity will disappear after heating. How to solve this problem, can you give me some advice to solve this problem? I have tested on PC chips to rule out fluorinated oils and surfactants.
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Are you using HFE7500 or FC40 as your oil phase? I would recommend using FC40 if you aren't already as it has a higher evaporation temperature. Since PDMS is permeable you can get some level of evaporation/fluid exchange across PDMS which can result in droplet coalescence (that you may not see on the PC chips).
If the chamber has inlet/outlet holes I would recommend plugging with tubing or something to reduce evaporation.
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I am working in control radiant flux of LEDs using a PWM controller. For this purpose I will use a MOSFET as a switch between the PWM chip and the LEDs. In this case should my mosfet operate in linear or saturation mode?
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Perhaps there some confusion regarding the terms here. The BJTs operate in 'saturation mode' when used as switch, usual definition being forward biasing condition of base-collector junction However, the 'saturation mode' for for MOSFETs means essentially current limiting mode, in 'switch mode' MOSFET is usually ohmic = 'linear mode'. Usually, PWM is used combined with switch mode.
However, one should note that LEDs should be current not voltage controlled. The easy way to implement it is using PWM controlled switch mode MOSFET and current limiting resistor. More sophisticated schemes may use PWM smoothing by using conductors, capacitors.
PWM controlled MOSFET based current source can also be considered but usually is more complicated and less efficient.
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Here you can talk and share your idea and experiences on network on chip simulators in any fields like Wire, Wireless, 2D, 3D, Photonic, etc.
feel free to ask questions and share ideas.
what is the best simulator in this field and what makes it better than others?
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Check the link provided below, where you can see a list of simulators for NoC.
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I am planning to do a lot of ChIPs following 'Chromatin techniques for plant cells Bowler 2004' and thus I am planning on stockpiling a lot of stock solutions and premade ChIP buffers. I'm having a very difficult time finding any consistent info on storage conditions for even individual solutions and whether they should be sterile and whether to do it by autoclave or filter. Multiple sources give conflicting information so I've tried to compile what I felt was the consensus below.
I was wondering if I could get any help confirming that the below information is correct and fill in the missing info?
A: typical storage condition/time in storage
B: storable at -20C?/time stable at -20C
C: storable at -80C? /time stable at -80C
D: light sensitive?
E: should be autoclaved?
F: should be filter sterilized?
G: special considerations
Stock Solutions
Formaldehyde 37% :
A: RT/2 yr B: Yes/indefinite C: Yes/indefinite D: NO E: NO (obviously) F: NO G: hazardous
Glycine 2M :
A: 4C/2 yr B: Yes/indefinite C: Yes/indefinite D: NO E: YES (autoclave or filter sterilize) F: YES
SUCROSE 2M :
A: RT/6m B: Yes/indefinite C: Yes/indefinite D: NO E: YES (autoclave as long as solution does not turn brown or filter sterilize) F: YES
Tris-Hcl pH 8 1M :
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
B-ME 14.3M:
A: RT/? B: Yes/indefinite C: Yes/indefinite D: ? E: NO F: NO G: hazardous
PMSF 0.2M :
A: -20C/? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO G: hazardous
MgCl2 1M:
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
Triton X 20%:
A: RT/indefinite B: ? C: ? D: NO E: NO F: NO
EDTA 0.5M:
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
20% SDS
A: RT/indefinite B: No/precipitates C: No/precipitates D: NO E: NO F: NO
NaCl 5M:
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
NaHCO3 :
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
NP-40:
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO
LiCl 4M: A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: NO ChIP Solutions Extraction Buffer 1 0.4 m Sucrose 10 mm Tris–HCl, pH 8.0 5 mM β-ME 0.1 
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
with sodium butyrate 5mM
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
Extraction Buffer 2 0.25 M Sucrose 10 mm Tris–HCl, pH 8.0 10 mM MgCl2 1% Triton X-100 5 mM β-ME
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
with sodium butyrate 5mM
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
Extraction Buffer 3 1.7 M Sucrose 10 mm Tris–HCl, pH 8.0 0.15% Triton X-100 2 mM MgCl2 5 mM BME
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
with sodium butyrate 5mM
A: 4C/6m? B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: YES
Nuclei Lysis Buffer 50 mM Tris–HCl, pH 8.0 10 mM EDTA 1% SDS
A: RT/1yr? B: Yes/indefinite C: Yes/indefinite D: NO E: YES F: YES
with sodium butyrate 5mm
A: RT/1yr? B: Yes/indefinite C: Yes/indefinite D: NO E: YES F: YES
ChIP dilution buffer 1.1% Triton X-100 1.2 mM EDTA 16.7 mM Tris–HCl, pH 8.0 167 mM NaCl
A: RT/1yr? B: Yes/indefinite C: Yes/indefinite D: NO E: YES F: YES
with sodium butyrate 5mm
A: RT/1yr? B: Yes/indefinite C: Yes/indefinite D: NO E: YES F: YES Elution buffer 1% SDS 0.1 m NaHCO3
A: RT/4hr B: Yes/indefinite C: Yes/indefinite D: NO E: NO F: ? Low salt wash buffer 150 mM NaCl 0.1% SDS 1% Triton X-100 2 mm EDTA 20 mm Tris–HCl, pH 8.0
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: ? F: ? High salt wash buffer 500 mM NaCl 0.1% SDS 1% Triton X-100 2 mm EDTA 20 mm Tris–HCl, pH 8.0
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: ? F: ?
LiCl wash buffer 0.25 M LiCl 1% NP-40 1% sodium deoxycholate 1 mM EDTA 10 mm Tris–HCl, pH 8.0
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: ? F: ?
TE buffer 10 mm Tris–HCl, pH 8.0 1 mM EDTA
A: RT/indefinite B: Yes/indefinite C: Yes/indefinite D: NO E: ? F: ?
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Dear AdAd,
that's kind of an unwieldy question. A lot of it is self-explanatory.
Let me see:
Formaldehyde 37% :
best to prepare fresh from paraformaldehyde
Glycine 2M :
filter sterilize, RT
SUCROSE 2M :
autoclave, RT
Tris-Hcl pH 8 1M :
autoclave, RT
B-ME 14.3M:
as given by supplier (4C), no need to sterilize
PMSF 0.2M :
make in isoprop, store at -20C, once in use at RT, decays in aqueous solutions within minutes!
MgCl2 1M:
autoclave, RT
Triton X 20%:
no need to do anything, RT
EDTA 0.5M:
autoclave, RT
20% SDS
come on, 20% SDS???? no need to do anything, nothing will grow
NaCl 5M:
autoclave, RT (try at 4C and you will see how you get very nice crystals....)
NaHCO3 :
prepare fresh, solutions decay by exchange with atmospheric CO2
NP-40:
no need to do anything, RT
LiCl 4M: autoclave, RT
For your working solutions it is best to prepare them fresh for each experiment with your stocks. That's easy and keeps contaminations away.
Good luck with your experiments.
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There is nothing worse for curb appearance than chipped and cracked concrete, particularly in steps. But you don’t have to put up with that blight any longer. By the result of weather changes exerted on an existing construction for prolonged time, it causes contraction and expansion those gradually results concrete steps cracks and break up with time.
Dear researchers, I would like to get expert opinions on how to fix this issue with,
optimum efficiency
low cost
prolonged sustainability .
Your answers are highly welcomed.
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En mi pais, hay material especial para tratar estos casos y otros mas; por ejemplo selladores, alquitranes, y otros.
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Hello Everyone, I am using ABAQUS & trying to simulate serrated chips while machining of Titanium alloy. But I am getting an error.
"An excessive temperature rate occurs in solving the heat transfer equations. This usually indicates that some elements are badly distorted or an error exists with model definition. You can check the temperature values and see if the distorted elements exist."
I am using ALE method. Any kind of suggestions will be highly appreciated.
Thank You
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Element deletion govened by J-C damage model.
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According to the number of Input and outputs ports of the FPGA chip we have, can we add (implement) any number of ethernet (100base) RJ45 and Fibre LC connector to the FPGA? What is the way to do that?
Also how can I also implement the following protocol into the FPGA; DNP3 , GOOSE and MODBUS?
which tools or libraries are using for these protocols on FPGA?
and what is best affordable cheap FPGA development board to buy with Ethernet Rj45 and Fibre LC connector SFP
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The paper in the link contains complete FPGA design and implementation on Xilinix VIRTIX 6 development board:
This paper introduces the field programmable gate array FPGA implementation of 1000BASE-X PHY Physical Layer for gigabit Ethernet over fiber optic cable. The implementation is achieved by developing VHDL model for all its building blocks including the physical coding sub layer, PCS, and the physical medium attachment, PMA. The VHDL code is simulated using XILINX ISE14.7 and synthesized on Xilinx Virtex6 FPGA chip. Measured results show that the designed and implemented Ethernet transceiver works successfully at 1.32 Gb/s, 2.5V supply with reduced power consumption.
There is even higher performance chips than Virtex 6 such as Verix 7.
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Hello, good morning. Our study is about ranking 30 blue chip stocks in the PSE. We want to know the validity of a ranking method called TOPSIS based on the ranking used by the Philippine Stock Exchange which is the VWAP method. I just want to ask if I can use Kendall tau b to know if the ranking given by the TOPSIS method is valid, based on the ranking given by the VWAP method? Because there are ties in the ranking between the two methods. Thank you! I hope you can help me with this.
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You may follow the following study to get some eflections on the uses and criticisms of Kendall's tau:
Lapata, M. (2006). Automatic evaluation of information ordering: Kendall's tau. Computational Linguistics, 32(4), 471-484.
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I've been working on differentiating C2C12 myoblasts into myotubes, growing these on glass coverslips as part of a microfluidic co-culture setup. They usually grow and differentiate quite well on plastic, and I've managed to differentiate them on glass coverslips in well plates with minimal delamination on occasion, with very gentle medium changes. I'm aware that I cannot prevent the delaminating on glass, but I am looking for ways to minimise this so I can have viable myotubes for long enough to perform my analyses (mostly ICC at this stage, but I'm also looking to eventually assess neuromuscular junction formation). #
So far, I've been coating my coverslips with laminin to delay delamination. More recently I even tried seeding an additional layer of C2C12s 2-4 days after the first, to act as a sort of 3D matrix, and this seemed to work well for me on the glass coverslips, but not as well in the microfluidics chips.
I've attached an image from ICC of my myotubes, after 7 days in differentiation medium (red is anti-myosin heavy chain, green is alpha-bungarotoxin to label acetylcholine receptors, and I use Hoechst as a nuclear stain). I'm not seeing as many myotubes as I'd like to between days 3-4, but by day 6-7 I'm seeing myotubes peel off and roll up into clumps like this.
I'd appreciate any advice at all, thanks!
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Isabella Bagdasarian hi thanks so much for this! I tried this recently but now have a few follow-up questions. What do you dilute your APTES in? I've tried another compound, DETA (N1-(3-Trimethoxysilylpropyl)diethylenetriamine), and used 1% in toluene at room temperature for 2 hours. Do you heat your solution? And for how long? Is this necessary, and do you use a hot plate for heating it?
I then did a wash in toluene before curing in the oven, but there was a lot of residue on my coverslips from the treatment. Unsure if I did something wrong, I mostly followed protocols I found in papers. Don't think we can do contact angle goniometry to see if it worked so mostly looking at how the cells respond and my 12-well plate was a mixed bag of results, but none of them lasted 7 days, never mind 2 weeks! Do you have a detailed protocol written for someone who's never done this sort of treatment before? Really appreciate the help, thanks so much again!
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Currently I am working on a microfluidic chip. An integral part of it should be a semi-permeable membrane which allows the flow of H2O and smaller ions (nothing bigger than a dinucleotide) but effectively blocks all DNA bigger than 12 nt.
For in situ polymerisation I use PEGDA (~MW = 700, others are in the shelf), a photostarter (Darocur 1153) and some water. Polymerisation starts with UV exposure and so far everything goes fine, the structure is where I need it to be and the mechanical stability also according to the requirement.
Is there a way to calculate the pore size upfront or a table where I can look up which composition leads to which result?
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You might find some answer about mesh size of PEGDA hydrogel (but with longer chains) in the following article:
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I am making microfluidic chips and trying to bond pmma to glass. what brand (model?) of psa do you recommend?
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Mahdi Ahmadi A couple of articles related to usage of polyimide PSA tape based on relative humidity of surrounding environment (Tan et al., 2008) and an alternative pmma bonding solution (Liga et al., 2016)
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Hey guys,
for an application I will need to a semipermeable membrane into a microfluidic chip. It should catch the DNA after an electrophoretic run.
The constraints are:
- MWCO at 2 kDa
- storable / not easily biodegradable
- low electrical resistance when trenched in a conductive solution
Do you have any suggestions? Are there hydrogels which are better suited than others?
Thanks
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The way that I have planned for my own work was to use a box like shoes' box by using lithography and inside box create two slites. The solid membrane would be prepared and then shift inside the site. Something like the old camera. The negative film would be replaced by the membrane. I have not tried it yet. I worry about the leakage but it seems there might be a way for it as well. I am not sure it works for soft membranes.
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I am recently using SPR to test the binding affinities between proteins and DNA aptamer. I used DNA aptamer as the analyte and I immobilized the His-tag proteins on the NTA chip surface. However, I got negative response in the Fc2-Fc1 curve. I assume this is due to the non-specific binding of negatively charged DNA and positively charged surface. I would like to know if that means it is impossible to test DNA aptamer using NTA chip? Or it is possible to avoid this non-specific binding?
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I agree with Vanessa Porkolab.
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What could be the effect of Waste Plastic irregular chips on the bearing capacity of compacted Soils? And If the Plastic is cut into regular shaped stirrups then how different will be the resulting improvement, if any?
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The plastics can increase the California Bearing ratio of the soil. You can try that.
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Hello,
The fist mode shape (Out of plane) of my MEMS chip occurs at 7.7 MHz. there is released cantilever in my chip and I would like to vibrate it to its resonance frequency. What method do you suggest to actuate the MEMS chip? If you have any product in your mind I appreciate that if you send me the link as well.
Best
Masoud
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Hi Masoud,
I'd propose you try the following two solutions:
1. Try to measure the resonance frequency of your cantilever using a laser interferometer without actuation. At 7.7 MHz, I'd expect there to be very little ambient noise and also that your cantilever has a pretty high Q-factor. This means that the cantilever will self-resonate at its resonance frequency, acting as a signal filter, which you will be able to observe.
2. As Vinod suggested, use a piezo element, firmly connected to the base of the cantilever chip. Connect the piezo element to a signal generator that will sweep the frequency in the range of interest.
Good luck with your experiment!
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I'm going to prepare a muscle frozen tissue sample ChIP with pan-acetylated H3 histone antibody. What inhibitors do I need to prevent spontaneous deacetylation of H3 and its degragation? What inhibitors do you use in this method?
Sincerely,
Kristina Sharlo, PhD
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You could use Trichostatin A which is a histone deacetylase (HDAC) inhibitor that inhibit the histone deactylase enzyme. Please refer to the articles attached for more information.
Best.
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I have some mice tissues stored in RNAlater. I was wondering if it is possible to do a ChIP or chromatin accessibility assay?
Thanks in advance!
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There's a PLoS One paper where ChIP was performed on RNALater-preserved tissue ( ) and since it is advertised as a preserving agent for DNA and proteins as well (https://www.researchgate.net/post/Western-blots-on-tissue-which-have-been-stored-in-RNAlater) I think it might work if your samples aren't too old
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Hi, I am working on a project to integrate a triple band chip antenna for wifi 6E to a wifi module. antenna is working on 2.4Ghz, 5.5 Ghz and 6.8 Ghz bands, is there any paper or reference of how I could do that and how to re-tune the antenna after integration, what problem I will face and how to deal with them...thanks.
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You need a matching network to match your antenna impedance to the circuit input impedance.
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we have a microarray assay with about 600 thousands markers. Illumina Infinium globalscreening-24v3.0 Bead Chip (48 samples) array. The results are like this "rs364728", how to transform the rs numbers of such a hughe package into concrete gene mutations ? Is there also possibility to add health conditions to concrete mutations ? Do you have any experience with this assay ? Thank you Dana Pokorna
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I agree with @Anita Tripathi
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Hi everyone,
I'm performing 2D Orthogonal turning of Ti6Al4V sample, although I'm able to see chip formation and run the model smoothly but I can't seem to get serrated chips which actually is my objective.
Cutting Speed : 50m/min, 120m/min & 240m/min.
Feed : 100 microns
Mesh element : CPE4RT
WP & Tool are both 2D planar shells.
I have attached the photos of my modelling herewith. Please help
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The following paper for your reference.
Zheng, Zhongpeng, Chenbing Ni, Yun Yang, Yuchao Bai, and Xin Jin. "Numerical Analysis of Serrated Chip Formation Mechanism with Johnson-Cook Parameters in Micro-Cutting of Ti6Al4V." Metals 11, no. 1 (2021): 102.
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Hello,
I am working on a mems chip. it is a released cantilever. the cantilever was on oxide and oxide has been etched by wet HF. however, we would like to fill beneath the cantilever with oxide. we etched some parts mistakenly and we need to fill them again. there is a schematic cross-section of the chip in the attachment. can we use PECVD? if not is there any way to do this?
thank you
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You can you use a solution which is composed of suspended silicon dioxide particles in a flying solvent. After covering the areas that to remain free from the oxide with adhesive film you can simply dip the structure in the solution after each dipping you need to dry it. You need to repeat the process sometimes to fill the gap as you want. There is such chemical solution that is normally used to deposit silicon dioxide on the silicon wafers to getter metals from them.
best wishes
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I have extracted DNA by a BigFish auto-extractor (BFMP02R) kit. The concentrations on a nanodrop are acceptable (30-120ng/ul) but the A260/A280 ratio is consistently low (1.2-1.5). The samples are intended for sequencing (may be some chip). Do I need to purify this with ethanol or will it go with sequencing in this shape? Or if can I change something in the wash or the lysis step? Add/remove something?
Thank you all in advance
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Do gel electrophoresis, If there is no proteins/impurities left in the well, proceed with sequencing otherwise try an additional step of washing.
You can also wash DNA with 70% ethanol before adding TE buffer to purify DNA.
Hope this will help.
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Hi folks! I am looking some examples of time temperature profiles folks have used to read TLD-100, 600 and 700 chips on a Teledyne 310 TLD reader.
There are a number of examples in the literature but people tend not to include the exact profile in the publications.
I know that it will likely be a simple scan from room temperature to 400C (or as close as the machine will go) with a heating rate of about 8 C / sec. But I would love to have a citation for this.
Thanks!
Tim
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Thanks Kindly!
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I wish to use CoolProp package in Octave in mac os (intel chip). I did steps mentioned from the following website: http://www.coolprop.org/coolprop/wrappers/Octave/index.html#octave. However, It is still not working. Can anyone please help me? Thank You.
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I agree with Hassan Nasser.
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Dear All We are working to make a liquid mixed with nano-sized quantum dots and a photocurable material ( Silica Colloid Vinyl Acrylate (SCVA) + DPPA / PETA + ACMO ) into micro droplets using a glass microfluidic chip. It is an experiment in which droplets are cured when exposed to UV on the channel, but the channel was blocked by accidentally irradiating UV.
Do you know of any chemical that can dissolve this cured photocurable material? Acetone, 3 mol NaOH, DMSO, etc. were already used, but the effect was insignificant.
Thank you very much Best Kim
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Try chloroform with ultrasonic bath.
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Hello everyone,
I'm looking for a software thats adapted for drawing complex designs of microchannels. Typically i have 2 inlets, 3 outlets and 100 microchannels in between. For the time being, i am using Clewin and Klayout, but i seek something more adapted for such complex features and this considerable number of channels. Do you have any recommendations. Do you know of a software to design such chips and able to export in .cif format that permits me to make my lithography masks ?
Thanks in advance,
Mohammad Baz
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Thank you Shayan Davani and Mohammad Taheripur for your recommendations, i will consider both.
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We have a Shimadzu ICP-MS 2030 that often suffers from what looks like a chipped torch after a few weeks of use. It typically only chips on one side of the opening facing the skimmer cones. The effects can be seen on the cone, as a white spot will typically appear across from the chipped area, rather than a solid ring shape that would appear through normal use.
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Hello Chris. Have you found an explanation or solution for your problem with the torch? I am facing similar problems currently whit the ICP-MS.
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I am trying to make collagen chip from type I marine collagen and infusing the same with different natural products for the treatment of periodontitis. How does one make the same?
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Dear Dr Malvika Singh,
I highly recommend you to use the protocol found in this article (in attached), to perform your technique.
The approach of Dr Sara Monsy Oommen is also very interesting.
Best wishes,
Sabri
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Dear All, I am trying to generate chip in Ansys workbench but chips are not coming out. Kindly help me to generate chip. Only red dots appeared near the cutting area.
Thanks in advance
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  • His-tagged protein immobilized a  on Ni coated NTA chip.
  • Both the reference channel and ligand bound channel shows binding with analyte (ribosome)
  • negative RU in the resultant curve.
  • Reference channel is Ni free.
  • Running buffer contains hepes,NaCl,EDTA,Mg(OAc)2
  • Both the ligand and analyte are prepared in the running buffer.
  • Reference channel was tried to be coated with BSA and still it's showing non-specific binding.
  • Tris buffer as running nuffer gave the same non-specific binding but the resultant curve gave positive RU. As suggested in some paper I am using Hepes Buffer now. If anyone has any idea please suggest me a way.
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Dear Mariana Amaral, it was an experiment I was trying to do a long time ago. Unfortunately, I had to move on to other experiments as it was taking a long time to standardize. However, the main problem I was facing was the magnesium ions in my running buffer as far as I can remember. Magnesium being a divalent cation was probably binding to the NTA chip surface, giving negative RU. I also had cobalt ion in the buffer containing the his-tagged protein. Besides, ribosomes being such a big molecule needed more standardization which I didn't pursue. However, as far as I can remember, all the above suggestions from other people help understand the crucial points one needs to follow in order to standardize the experiment which I left inbetween.
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I am currently working on a project that requires the prediction of the oblique shear angle for an oblique cutting. The numerical iteration should continue until the chip flow angle converges within 10^-14
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I need too. If you got the answer, could you please to share to me?
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Dear All
I would like to bond my PDMS chip to a glass but the size of my chip will be greater than a usual glass slide. Therefore, I need to bond it to a larger glass. Is that possible? Or can it be bonded to another PDMS layer instead?
Thank you very much.
Best Regards
Su
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Is it right to classify the consumption of wood chips among renewable energy sources? According to EUROSTAT, its consumption is the fastest growing type from renewable sources ...
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Ladislav Rozenský Biogas generated from any type of organic feedstock can be consumed and it can be a game changer...
Already Europe os doing and similarly UK and USA are following and now India is following the same
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Hi, I was preparing samples for Chip-seq from murine bone marrow-derived cell lines using Active Motif's CHIP-IT Express magnetic CHIP kit. I started with 10-15M cells and followed the protocol at every step; however, even my input DNA samples only had <7ng/uL of DNA (quantified by Qubit) after DNA purification, which is a really low yield considering how many cells I used. Any possibilities that could lead to low DNA yield? Could it be a result of insufficient homogenization?
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I agree with the previous suggestion. The lysis might not be enough. Please vortex prior to centrifugation step. You can store the DNA for overnight -20 . Next day you can go for purification step. By doing this, there are chances for increased precipitation of the DNA. Hope this helps.
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Hi everyone,
Since new semiconductors with complex tomography emergy, it usually needs hundreds of processes such as lithography, etch, et al. from an unpattered wafer to an enveloped chip. It seems that the metrology and inpection play an important role in the process and yield control. Also, there are various devices for critical dimension metrology and defect inspection. However, with respcet to efficiency,I wonder which processes really need critical dimension metrologies and defect inspections?
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Hello Renju Peng,
Each production step usually under control by non-invasion methods. Main goal to save money in case if something goes wrong (e.g. issue during film deposition). To prevent customer device damage special structures can be used like rectangle shape for each type of films to have big area for measurement (not routing from IC). Generally better to scratch wafer before metallization if earlier processes went wrong like diffusion for wafer Si itself.
Best Regards,
Denis
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ChIP qPCR analysis
Hi everyone, I perfomed a ChIP experiment and I made a qPCR to check my results. However I'm experiencing some trouble in analizing my results through percent input method. I got the following Cts for my samples: INPUT Ct=24; IP Ct=20; MOCK Ct=33. Do you have any suggestion on how to analyze my data?
Thank you for your help
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Usually, two methods used in ChIP-qPCR quantification: percent input and fold enrichment. The calculation and comparison are in the following website: https://www.thermofisher.com/hk/en/home/life-science/epigenetics-noncoding-rna-research/chromatin-remodeling/chromatin-immunoprecipitation-chip/chip-analysis.html
Hope it helps.
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A. Global semiconductor chip shortage
B. Government regulation of internal combustion engine powered vehicles
C. Disruption from digital start-ups
D. Trade tensions between USA & China
E. Lasting economic impact left from Covid-19
F. Other (please comment)
I am collecting research for an Engineering Management (MSc) dissertation.
Any responses are greatly appreciated.
Thank you,
Joseph Dodd
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On the basis of delays in product deliveries, clearly the first point. As regard the economic investment, the second one.
Fredriksson, G., Roth, A., Tagliapietra, S., & Veugelers, R. (2018). Is the European automotive industry ready for the global electric vehicle revolution? (No. 2018/26). Bruegel Policy Contribution.
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The supplier has provided Lithium chips in the one-time opening can made of Al. But Al containers are not easily available. Please suggest.
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Ritesh Yadava , HDPE will do the job for the electrolyte. If you are using additives such as FEC then you need to be careful. Since FEC is light sensitive, then you need to wrap the nalgene bottle with aluminium foil. For lithium you can use store inside a glass bottle, make sure you can tighten it very well, incase the glovebox get contaminated it will still protect your lithium for a short period of time until you purge and rectify the problem. Hope this helps!
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Hi,
I want to do ChIP on jurkat cells but with my protocole I can't fragment the chromatine enough (fragment now are between 3000 and 1000 bp), even at 1h of sonication....
Do you have advice? (sonication, buffer, ...)
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Sorry. You have to draw the DNA through a needle. Which gauge, I do not know. It is an old method.
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I was under the impression that CUT&RUN's benefits came primarily from reducing cell input count. But with (not so recent) improvements in ChIP such as Nano-ChIP and LinDa-Chip, the latter of which brings down cell count to 10k cells (but let's take 50k for discussion), what reasons are there to really choose one method over the other?
What are the advantages of each method now that cell counts needed are much lower for ChIP sequencing, are they both equally good or are there reasons to choose one over the other?
-Jelle
Edit1: references of LinDA and Nano-ChIP
Shankaranarayanan, Pattabhiraman, et al. "Single-tube linear DNA amplification (LinDA) for robust ChIP-seq." Nature methods 8.7 (2011): 565-567.
Adli, Mazhar, and Bradley E. Bernstein. "Whole-genome chromatin profiling from limited numbers of cells using nano-ChIP-seq." Nature protocols 6.10 (2011): 1656-1668.
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Dear CK Gomathy, thank you for your answer, but I feel this doesn't really adress any of my questions. All the things you have mentioned can be done by Cut&Run, my question is why we would want to choose one over the other?
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Dear All
I aim to design a microfluidic chip with a quite long channel with rectangle spiral structure and inside cells will not be seeded but instead, flowing for an extended period of time (1-2 days). Therefore, I need a large surface area to fit such structure. What is the maximum size I can achieve? Or basically is it the maximum surface area achievable with 4-inch wafer?
Thank you very much!
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Hello Su,
You're welcome. Good question. Choosing the material for your substrate depends on several factors, for example, equipment in your institute, minimum feature of the designed channels, required thickness, hydrophilicity or hydrophobicity of chemicals used in the drug testing....
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Currently, I am generating agarose dropets on a microfluidic chip using mineral oil with span 80 for the carrier (oil) phase. The next step is to break the emulsion and free the agarose beads. So far I found that this is done by repeated washing (up to 7 times) with mineral oil without Span 80. Can anyone suggest a faster, more efficient way to break the emulsion and free the agarose beads?
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Hello,
I am wondering how you made the agarose droplets to begin with? I bought ultra low gelling agarose from sigma aldrich and it is gelling at room temperature:
Wondering if you dissolved the agarose in a special way with different pH?
Thanks,
Karl
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I'm trying to simulate the formation of segmented chips while performing turning of Ti6Al4V, I don't know why my mesh is getting distorted eventhough I'm using a fine one!!!
My wp consists of 3 zones, 1) uncut chip thickness zone where segmented chips are supposed to form, 2) Thin sacrificial layer, 3) Machined zone which results when chips have been formed. For 1 & 3 I've only applied material with JC plasticity no JC damage, for 2 I've assigned JC damge with evolution as well.
Please help.
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Amal Rai Unfortunately, I'm not familiar with Abacus. I saw this approach in many papers. Recently I'm trying to adopt the Discrete Element Method to simulate the FSW process.
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Hi,
I need some chips with interdigitates electrodes and microheater on silicon for research, do you know where can I buy it?
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Hi there, blood sugar level can be detected by bio chips based on LOC(lab on a chip) methods. I am wondering whether Oxytocin can be detected in a same way directly by biochips.
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This is a fascinating question. Unfortunately, I am not an expert in this field. However, I believe this publication may provide you some useful information. The links is:
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We have a BI4500 SPR, by which we can generate very good data for BSA/anti-BSA antibody and biotin-DNA/cDNA interaction.
Recently we try to use this machine to study ssDNA aptamer/protein interaction. We immobilized protein by CM chip, NTA chip or avidin chip, but can't observe any binding with the aptamer. We also use avidin chip to immobilize the biotin-aptamer, still can't see any binding with the protein. However, we successfully observed good binding if we use flow cytometry or fluorescent imaging for this aptamer and protein. This is very strange!!! Could you pls give me some suggestions?
Thanks.
Shawn
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Good luck, Shawn!
In case you have success, I would like to hear from you.
Yours,
Hans
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i am trying to find KD of aptamer as the analyte and protein. I'm using a CM sensor chip. I'm using PBS with tween 0.005% and 1 mM MgCl2 as RB. Normally I don't use ethanolamine to block the channel because aptamers don't have primary amine that can bind to the activated COO- and also we observed good binding without EA. we thought EA blocks everything and so we didn't use EA. with EA we couldn't generate any results. recently we got a good KD and it was pMol range. so it is too good to be true for this aptamer. when we checking specific binding, we realized there was some mistake in this experiment. so we activated the reference channel and injected the aptamer. we saw the binding, but that is not a non-specific binding. does someone have an idea about yhis? if that please help with this.
we assumed there should be a kind of positive charge interaction and that should be some kind of contaminants of the CM chip. it is a new CM chip. does anyone know any cleaning procedure for CM chip?
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Vanessa,
We generated SA chip by ourselves and successfully immobilize biotin-aptamer on the chip, but still, we can't observe the binding for the target protein.
This is very strange, so we did several control experiments: 1) we successfully observed the interaction between biotin-ssDNA and its cDNA; 2) we confirmed that the aptamer can bind the protein by flow cytometry and fluorescent microscope.
Could you pls give some advice?
Thanks.
XT
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Anyone aware of a supplier that can deliver waffle trays for storage of diced silicon chips? A supplier that does not require that you by +2000 trays.
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Ask for a sample of 10 and the pricing for 1 million... They'll call you 'Sir'...
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Hello all,
I am interested in fabricating a particular design for a microfluidic chip used for particle sorting. Would you please kindly let me know where I can find a lab that offers a microfabrication service for a customized design in Germany, and what the expected costs are, and how much time it takes for delivery.
Kind regards.
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Hi Abdulkarem Odhah I don't think that Bartels does and for Microfluidic shop (I don't know) you can write a request and they'll tell you.
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hello, am working on microfluidic device and my chip is block with sodium alginate and ?lactoferrin gel mixture can anyone suggest for me solvent that help melt this gel in my chips
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From the picture, it seems that you willl need to replace the chip.
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How to determine the Reducing and non reducing sugar in dry apple chips
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Hello sir, I am a university sudent. CFD analysis on pin fin water block is the problem statement iam working on.
i completed the entire problem and in the contour region iam getting the tempereture results as shown in the image below. where the minimum and maximum temperatures are too low and too high respectively. help me out in finding the error. i need to find out the resultant temperature of the CPU chip after the fluid flow through it. waiting for responce sir. thanking you.
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The image is quite pixelated and it is quite hard to read the values on the color bar. You need to provide more information on the boundary conditions used. Without knowing about the case setup and so on, I would say that this is either a result of the flow behaviour having local maxima and minima.
Although the flow has converged, the solution for the temperature equation must not have converged or must have diverged.