Plasma - Science topic

The number of participants and their demographics could be a factor. Race and differences in cardiometabolic issues might be influencing the differences miRNA expression. Try look at the DEGs associated with the miRNAs that didn't match up, this will give you more insight into what's happening. It could be technical differences like choice of housekeeping genes, normalization process.

This question is difficult to answer If we talk about single streamers, then the resistance is 500 kOhm for a 10 cm long streamer. Approximately the same resistance has a 5 cm long streamer. If you are interested in this question in more detail, then you can find the answers in our article PROBE MEASUREMENTS OF PARAMETERS OF STREAMERS OF NANOSECOND FREQUENCY CROWN DISCHARGE

Ponizovsky A.Z., Gosteev S.G.

Physics of Atomic Nuclei. 2017. Vol. 80. No. 11. pp. 1704-1710.

If you have any questions, I will be happy to answer them my address sasha_laron@mail.ru Sincerely Alexander Ponizovskiy

Thank you Anupriya, the antibody concentration is 1:50000, I have also tested 1: 10000(attached picture last two bands) Strep-HRP(20ml, completely cover the blot) but I think the result is similar only more intensed. I also prepared fresh antibody each time.

The DPPH (2,2-Diphenyl-1-pricylhydrazyl) method can be used as a chemical method for determining the content of free radicals (see attached article).

Hi Donis, did you figure out this problem? The same thing happened in our study. We tested mice plasma with novusbio elisa kit.

You have low density plasma (the plasma power is too low) or the distance from the substrate to the target is too high. As my colleagues Jürgen Weippert and Jignesh G. Hirpara were saying look for plasma reflection back to the magnetron or verify if the substrate does not charge itself electrically.

Hello Oleg Agamalov,

At the moment, I don't have any work related to the magnetic reconnection converter. But, in the near future, I think it will be part of my research.

Best regards.

I think you're saying you're getting an unacceptably high background reading from your negative control. It's good that you're concerned about this problem. It is common, especially for human blood samples.

If you are using human blood, be sure to take proper bloodborne pathogen precautions, including your vaccinations and PPE.

Consider using serum rather than plasma. And for human samples you should heat the sera to kill viruses.

You should be diluting your samples in blocker solution. For ELISA you probably can start with a 1:1000 dilution and work toward 1:1000000.

You can reduce non-specific binding by adding things to your blocker solution. Starting with a base of PBS, blockers usually contain a protein like casein or albumin at 0.1% (1 mg/mL) and a nonionic detergent like Tween at 0.01%. You can try using a different protein. You can safely increase the protein 10-fold. Depending on the ELISA format, you may want to include 1% control serum in your diluent and blocker as a blocker protein. Increasing the detergent (up to 10x) may decrease your overall signal, but it may increase signal/noise ratio. And sometimes it helps to add 0.1M glycine.

Hi Christian,

interesting question - which wavelength would you need for that 13.5 nm, or even shorter?

I am pretty sure that you can do that but I think there are much better plasmas than tin - feel free to drop me a PM, if you want to discuss.

cheers

Johannes

Dear Prof. Saima Mukhtar

The kits that are labeled as “research purpose only” are exempt from regulatory requirements and approvals that would be needed for clinical diagnosis or patient management. They are supposed to be used only in research and discovery work. Since there are no guidelines for such “research purpose only” kits, the extent of manufacturing quality compliance, assay characterization and documentation will vary considerably across different vendors.

On the other hand, for the diagnostic kits, pre-market regulatory clearance is required by the FDA. The review of analytical and clinical validity of a diagnostic kit is done prior to marketing and thereby its use on patient specimens in clinical diagnosis where in the diagnostic kit identifies, measures, or predicts the presence or absence of a clinical condition or predisposition in a patient.

The “research purpose only” kits which have no regulatory compliance, no manufacturer dependent specifications, the kit inserts being provided have variable degrees of information, and a wide variety of sample matrix (e.g., animal/human fluids, tissues, culture media) being present, cannot be used for diagnostic purpose.

So, I feel you should not make an attempt to translate the results obtained from kit labelled as “research purpose only” to diagnostic values.

Best.

Thank you so much. Sure I will look after the software too@Bharat Singh Rawat

using zeeman correct

It depends on your analyte and assay mechanism.

Hemolytic samples can be measured photometrically (by measuring free hemoglobin) and can be prevented by having systems and recommendations in place.

Icteric samples can be identified photometrically at an absorbance of 480 nm and 505 nm. Resolving icterus can be accomplished by sample dilution to reduce icterus below the interference threshold.

Lipemic samples is most often detected indirectly as a measure of sample turbidity at a light absorbance of 570 nm and 660 nm. We can resolve this inference by sample ultracentrifugation.

Hi Sepideh,

Biomatik offers an extensive list of over 38,000 high-quality ELISA Kits. Feel free to check out our Insulin and IGF-1 kits here: https://www.biomatik.com/search-results-page?q=%20IGF-1&page=1&rb_categories=ELISA%20Kits&rb_custom_field_4af649a635c21db41b73eafb1def22da=Mouse

Best regards,

Biomatik Team

Yetoka Swu Thank you so much for the clarification. As mentioned by you in the last paragraph, can you suggest some papers/works where both the scenarios are addressed explicitly.

Use -20 degree C at lest.

I use the program Prism GraphPad for that. The data used is only the concentration level of the standard curve and the optical density levels. Then I interpolate the values for the estimation of the concentration of the protein in my samples. After selecting the interpolate option, a parameters window is open, where I select the 4 parameter logistic regression.

I hope this can help you.

You could call this a "smoldering hot topic" in sputtering - everybody asks for it at some point but the answers are often disappointing.

In principle, you can measure the formal temperature of a plasma by simple pyrometry and pretend that that's the temperature everything has. Of course you have to check with OES references that none of the characteristic wavelengths in the plasma coincides with the wavelengths your pyrometer uses.

Then you have a temperature which you can write on a report and pretend to have done it. However, the energy distributions in sputtering plasmas may be highly inhomogenous, especially when you come close to a biased substrate. You could even debate if the states inside there are homogenous enough so that the mere concept of a temperature makes sense in there.

There are concepts for calculating the energy distributions of particles impinging on the substrate from the plasma, but everything I've seen so far had major caveats on it.

Thank you so much dear Sohail Mumtaz for your answer.

Here is the DOI to my paper developing the wavenumber spectrum from wavelet analysis: https://pubs.aip.org/aip/pop/article/30/8/082303/2906229/Estimates-of-the-wavenumber-wavelet-power-spectrum

Zulfiqar Ahmad : Thanks for your good question - if your first author is located in a country outside the G20 countries (i.e. your institution is), there won't be any fees automatically but if you want to be sure that the fees are waived, you can leave me a short notice in the submission system, to remind me that we talked about this here on RG.

So, in any case there won't be any fees for you, if you make a submission to JTSP.

I hope, this answer helps and please feel free to share this information also with your colleagues, who might be interested.

thx

Try discus the topic with Lixoft they have amazing software and free of charge academic version probably is a possible option. Link below:

Hi have you tried Qiazol reagent ? it is also good for isolating RNA from lipid bound tissues.

I used the AssayGenie protocol and worked perfectly for me. There are 2 codes for mouse IL6. You should be able to find the protocols in the websites.

If you still have trouble, I can send you pictures of the protocol

Hi Atchara, My name is Eilysh and I am a Product Manager at STEMCELL Technologies.

Thank you for your question. To collect plasma for further analysis, use SepMate as instructed and then pipette off as much plasma as required after centrifugation.  Please remember to pipette off the plasma before pouring the sample containing PBMCs into a new tube. Learn more about the protocol here: https://www.stemcell.com/products/brands/sepmate.html.

I hope this helps! If you have any more questions, feel free to email us at techsupport@stemcell.com.

Kind regards, Eilysh

Hi Claudia

I am rather intrigued. What's the purpose of developing antibody against bcr-abl chimeric protein? Is it for therapeutics? This protein is intracellular and its action is intra-nuclear. How will a therapeutic antibody against this protein get to the target and neutralise it? TKI are very smart molecules capable of inhibiting transcription. How will anti bcr-abl antibody will work? I would be very interested to know. Best wishes. Siva

i understand this is a simple procedure i did not think about . thank you so much

The choice of diluent for preparing standards and experimental samples in analytical methods can depend on several factors, including the solubility of the analyte, compatibility with the analysis technique, and the desired matrix for calibration.

Here are some reasons why diluting standards with plasma or other biological matrices may be preferred over diluting them with methanol/water to match the experimental samples:

It's important to note that the choice of diluent depends on the specific analytical method, the analyte being measured, and the overall objective of the analysis. Methanol/water dilution may still be appropriate for some cases where the analyte is stable and soluble in that solvent mixture and if matrix effects are not a significant concern.

Ultimately, the decision on the diluent should be made based on method requirements, validation considerations, and the desired accuracy and reliability of the results. It is recommended to consult the specific method or assay protocol, as well as relevant literature or guidelines, for guidance on appropriate dilution procedures for your specific analysis.

Hello dear i am doing the same experiment targetting mRNA in plasma, if you got any possible solution please let me know how you did that before 2 years in 2021?? please i am requesting you either answer here or email me wafaawajid3617@gmail.com Ruth Stuckey

If you use plasma-enhanced MBE as in

you have, of course, a plasma inside, but that is a niche version. Most MBEs don't use a plasma.

When performing an ELISA assay, the dilution of the sample depends on various factors, including the concentration of the analyte of interest and the detection range of the ELISA kit. In the case of mouse IL-6 ELISA assay, the suggested starting point for dilution is to use PBS supplemented with 10-50% animal serum. However, the specific percentage within this range should be optimized for your particular samples.

To decide between using 10% or 50% serum, you can start with a pilot experiment to determine the optimal dilution for your samples. Here's a general approach you can follow:

The optimal dilution can vary depending on the concentration of IL-6 in your specific samples. It's important to consider the detection range of the ELISA kit and ensure that your diluted samples fall within that range for accurate measurement.

Hey, Gil.

an extra question. 20 mM CaCl2 is at final concetration (i.e. 20mM into the plasma) or is the concetration of the working solution (before adding to the plasma)? If is the second option, what is the proportion of CaCl2 and plasma?

thanx a lot in advance for your answer!

best regards

Ivo

"CROSSFIRE FOR MOBILITY & RELIABILITY, THAT IS PREMIERSHIP & GUARANTEED ARE ALL ABOUT" Elizabeth Holmes

Grzegorz

Use phosphate electrolyte.

Dr. K

Thansk a lot. Eventually the problem was not the BSA. I have a problem working with glass tubes. The reaction to remove the cholesterol (KOH 1 M) is very strong and it seems to affect to the glass...If I carry out the reaction in eppendorf I do not have problems.

Thanks a lot for your time!

To determine the exact amount of total polyphenols in plasma using Gallic acid as a standard, you can use a colorimetric method called the Folin-Ciocalteu assay. Here are the general steps involved in the procedure:

Note that this method measures the total polyphenol content in the plasma sample, not the individual polyphenols. Also, the accuracy of the measurement can be affected by various factors such as the presence of interfering substances and the stability of the polyphenols in the sample. Therefore, it's important to carefully validate the method and consider its limitations.

RIE is the name of the process. In normal sputtering process we physically remove atoms from the substrate surface by bombarding them by using energetic ions. But, if we choose the gas chemistry in such a way that these ions are also capable of reacting chemically with the substrate (and form volatile etch products) then the rate of etching can be increased drastically. This is RIE. RIE reaction chambers can be made in several geometries (cylindrical, parallel plate) but the parallel plate reactor is most common. In a parallel plate reactor the upper electrode is grounded, and the lower electrode is connected to a RF generator. This is the basic CCP configuration common in old systems. The plasma is created due to the large potential between the two electrodes and the oscillating field. Please read the wikipedia page for more details on how plasma is struck. However, in this setup, both the density of plasma (ions/area) and the kinetic energy of the ions are dependent on the power supplied to the lower electrode and cannot be changed independently. The solution to this is the ICP setup, where a seperate coil is attached above the upper eletrode and a seperate RF power is applied to the coil. This way the gas is first intoduced to this ICP coil and plasma is struck there, so that we can control the plasma density by changing the power supplied to the ICP coil. Then the ions are introduced into the chamber and acclerated towards the lower electrode due to the biasing voltage acheived by applying power to the lower electrode. This bias voltage controls the kinetic energy of the ions.

Try defibrinating or adding citrate.

yes the rf power to the target should be delivered and there should be perfect impedance matching. This will give you a good value of DC self bias voltage, and effective sputtering as Mr. Jurgen has mentioned.

You dont apply DC bias voltage,

It is a self biased developed getting developed on its own.

Well sputtering pressure can influence the self bias DC voltage. Try varying the sputtering, but always ensure 0W reflected power.

1. What ever target you are using can be problematic sometimes. There seems to be a serious impedance mismatch problem between your target and rf power supply.

1a. There can be an air gap between your target and electrode.

2. Try putting a simple Copper target of 2 to 3 mm thickness (parallel plate), and see if you are able to get a dense plasma and increase your power beyond 50 W.

3. 50 W is a lot of power on 2 inch target diameter metal target to give a fantastic deposition rate.

Dear Sir you may try the protein separation with foam fractionation coloumn. it would suppose to help you

Gotcha.

Umph. And Ulysses data are too old and Cluster's are in the wrong place...

I'd drop Dr. Heyner a line:

10-500 ng/mL (or 0.1-5 nM) Musa Ibne Mannan

What are you looking for?

If you use whole blood, you'll actually get meaningful amounts of mRNA, because blood contains transcriptionally-active cells.

if you use serum, you kinda won't, because serum is a cell free fluid. You'll maybe have trace mRNAs released from damaged cells, but these will be few in number and thus low in abundance.

Serum will, however, be rich in miRs, both released from damaged cells and as constitutive components of the circulatory milieu.

If you are interested in serum microRNAs, serum is fine. If you're interested in mRNAs in microvesicles/exosomes, then serum is also probably fine, but if you're interested in anything else, then I'd say: use blood.

With respect to the very first question – leaving aside other possible issues, the main problem of the chosen approach of cosputtering arises from assumption, that to reach mentioned composition the deposition rates of both cathodes are equivalent when equivalent power is applied. This assumption is incorrect in general.

There is already a discussion dedicated to this problem (How to find the sputtering power and time of sputtering of individual targets to make a desired alloy composition using co-sputtering technique? | ResearchGate) where some possible sources are mentioned.

Dear,

What is your target price?

To answer this question, further readings into studies conducted to find out whether or not salivary matrixes could be an alternative for antiepileptic TDM would be beneficial. In terms of traditional linelozoid TDM, salivary matrix can be used as an alternative however, there are limitations on the uage of saliva universally for TDM, such as is the case of moniflaxin TDM, which does not support saliva, according to the study (Therapeutic drug monitoring using saliva as matrix: an opportunity for linezolid, but challenge for moxifloxacin, Elsen et al., 2019).

Thanks for the answer. I also think same.

Serum is the liquid remaining after the blood has clotted. Plasma is the liquid after the blood cells have been removed and clotting has been prevented. Unless you are specifically interested in serum, I would consider plasma to be the relevant physiological fluid. I have seen transthyretin described as a plasma protein, and this makes more physiological sense to me than describing it as a serum protein.

Hi

The yield obtained depends on technique used after the whole blood sample was gotten from -80. The sample should be thawed quickly on Aluminium blocks at room temperature and further processed using the normal technique your lab adopted. Avoid usage of water bath as it takes long time and leads to more RNA Degradation, this improves RNA quality and yield .

Yh you can, once the route of collection and the processes are right

Are you Tom molecule of the atom change it with toxic acidity. Hydrochloric acid is poisoning the system. And affecting the molecules and the atoms. I am an atomic engineer for an atom physics. I could tell you more but I've told you enough already. Crisis and emergency alert http://youtu.be/Ng1-KJueYiU Time for the people to stand together to bypass, help us build the bypass. We have the foundation's know

Yes, the tubes had heparin...

The pH of my ELISAs protocols buffer are the following ones:

On predicting the pH of pure sodium acetate aq. sol.; see: https://www.researchgate.net/post/Can_we_keep_the_pH_of_3M_Sodium_Acetate_around_7_instead_45-5_while_using_this_in_DNA_extraction_from_whole_blood

Thank you!

Fluctuations in discharge current during a plasma discharge are common and generally depend on type of discharge mode, plasma impedance and impedance of generator along with the length of the leads used with generator. These fluctuations can be due to:

1. Impedance mismatch between generator and plasma results in reflection of enenrgy back to source. In such cases in coming and reflected signals form multiple harmonics of applied signals some times appearing as noise. A matching network is highly recommended in this case.

2. Filamentary discharge where rapid oscillations of space charge under applied pulsating voltage results in such type of discharge current signals.

3. Smaller leads used with oscilloscope.

Sufficiently longer leads not only separate the in coming and reflected signals but also reduce noise particularly at lower applied frequency.

Hi Kathrin,

we have not tried commercially available reference materials so far, however we prepared our own and used them in various methods including single EV flow cytometry (published in ) and bead-based flow cytometry ( ).

More recently we also used fluorescently tagged EVs as reference material to evaluate EV stability and storage conditions in various methods including simple bulk plate readers ( )

You don't need huge amounts for any of those methods, if you still need some of those EVs I'm happy to share those - just email me :)

Otherwise fully agree there is a high need for more rigorously characterized EV reference materials for different purposes - I know we and some other labs are working on that :)

Hope that helps,

best wishes,

Andre

Your dna is probably below visual detection even if it were a single size band on the gel but plasma dna is randonly degraded and small so is spread over a range of sizes on the gel so no one size will be visible. Seeing dna is not very important if the next step of your research is a pcr amplification then go ahead and amplify and it will then produce visible bands

Thank you all for your answers,

but the electrons are non-isothermal, i.e., represented by a vortex-like distribution function. In other words, there are free and trapped electrons, and the last ones are trapped in the electrostatic potential trough.

Dear Adam M G Burgess ,

My pleasure

perhaps the problem is the strength/type of your acid solution? If you are using a strong oxidising acid like nitric acid and concentration is high then this will consume the reductant (NaBH4) so that not enough reductant is available to reduce the As quickly. I have seen this problem in the past for mercury hydride systems.

Langmuir probe works in plasma exposed to magnetic field?

My plasma generates high density fusions, probe must withstands that

Dear Danielle,

I have not the experience with plasma freezing, but I isolated total RNA from culture cells and blood. For both the most important is to freezing and storage the samples at -80 degree, because RNA is very unstable. In the case of blood we used the specialized kit for blood freezing, but I recommend carry out the isolation with unfreezing sample (as soon as possible).

Hi! Once again, thank you for the reply! I have never published before, that's why I was asking :D

First you should determine fat content in butter. The rest is butter plasma. Example: fat content is 80% (butter plasma is 20%) and total acidity is 0.05%. The acidity of butter plasma is 100*0.05/20 = 0.25%

How long ago was your last chamber cleaning? This might be due to redeposited material from the walls crumbling off and flying around in the plasma.

It can vary depending on which kit you are trying to use.

But the range is anywhere between 50uL-5000uL.

Thanks Malcolm

This is unfortunately what I thought, but thank you for confirming! Back to the lab.

the cell become shrink and the chromatin condensed

Disassembly of organelles.

Fragmentation of cells and Degradation of DNA, ill defined surface of the cell

Hi Carine

Plasma Hb can be accurately measured in samples with other anticoagulants, e.g. EDTA, citrate, etc. Lithium heparin is just one of many anticoagulants.

In clinical labs, plasma Hb is usually measured on a biochemistry analyzer instrument, along with several other analytes using the same sample. Heparin is often recommended by the instrument manufacturer because it is compatible with some of the other analytes also being tested in the sample, such as pH. Choice of anticoagulant is therefore dictated by these other circumstances, not by a specific biological requirement of the plasma Hb assay.

The article that you referred to was actually comparing 6 different assays/analyzers, not different anticoagulants. The important consideration is sample consistency - so use the same anticoagulant (and blood processing/handling conditions) for your entire set of experiments.

All RNA extraction and library preparation techniques studied consistently detected small RNAs, but several miRNAs had considerably variable amounts. The relative relevance of minimizing the amount of total sequencing required, finding uncommon miRNAs, or absolute quantification should be considered while choosing the best methodology.

Fresh frozen plasma to store in approved freezers at less than -30°C. It is thawed just before use (a process which takes up to 30 minutes) and once thawed, must be infused within 24 hours if kept at 4°C (or 4 hours if kept at room temperature)

A reliable plasma marker of platelet activation: does it exist?

Yes, I know. We do this with the help of low-temperature "cold" plasma =

V. Tikhonov, etc. "The Low-Cost Microwave Source of Non- Thermal Plasma," 2020 7th International Congress on Energy Fluxes and Radiation Effects (EFRE), Tomsk, Russia, 2020, pp. 596-599, doi: 10.1109/EFRE47760.2020.9242089.