Seeds - Science topic

Misbah Naz

Thanks for your valuable suggestions.

Alex Ignatov

Thanks for your valuable answer.

Hi Sam

Thank you so much for the answer.

The DOI: may help you

Yes express TLR-4 and LPS increase it. I know that FBS has sCD14 and is necessary for LPS but in my previous tests I used FBS and I didn't get results so according to an article I eliminated FBS.

Nils Wagner Hello sir, I am trying to add but am not able to add..Any other way to share sir.

have you checked for mycoplasma?

See also an earlier discussion on this topic: (5) 3,3-Diaminobenzidine (DAB) solution making for seed staining? | ResearchGate

Hello Navodya Silva

To achieve an even distribution of cells in the well for a 24-well plate, I make use of figure eight-like movement, while one may also use a cross-like movement of the plate. Both these techniques lead to a better distribution of cells in the well.

However, for wells of smaller diameter such as the wells of a 96-well plate, the movement of the liquid is less, leading to an uneven distribution of cells. An effective way to circumvent this effect during cell seeding would be to first dilute the cell suspension to the desired concentration in a tube and then pipette the final volume into the well. Do not add the cell suspension to the well already containing the media for dilution.

Moreover, do not shake the plate after seeding the cells as this shaking movement may move the cells to the edge of the well. A little vibration while walking with the plate from the laminar hood to the CO2 incubator may also cause uneven distribution of cells in the well.

So, I suggest after having diluted the cell suspension to the concentration you desire, and after having pipetted the final volume into the well, you may use a tray to transfer the 96-well plate form the biosafety cabinet to the CO2 incubator.

Best.

Hey!

I am not sure if this will work for you or not, but in general, whenever I seed cells(be it primary or a cell line) I always move my flask in the shape of the number 8 (Keeping this in mind: be gentle ). This will distribute your cells uniformly throughout the flask.

You can also try coating your culture flask first as per your cell requirement. This will help with the proper adherence of cells.

Avoid moving your flask too quickly or doing harsh pipetting.

Dear Peter Breuer, Thank you for your attention.

I mean, the runs are initialized the same (data starting and ending times are the same and fixed to a predefined value without adding any random value).

The point is that using the RNG in the same uniform interval leads to different results on the two versions of Ubuntu. For example, I have used the following command in my code.

In Ubuntu 14.04, for different executions in the same conditions, the above command does not change the nodes local topology formation, resulting in the same outcomes.

However, even by changing the interval and making it smaller in Ubuntu 16.04, the same command leads to a change in the formation of the list of neighbors and, as a result, a difference in different executions of the same code under the same conditions.

Try again, but with only viable seeds. You can test for viability using the flotation test. Seeds that sink in water are viable. And try longer too :)

Several studies claimed its toxicity So in conclusion go through this review paper, which may solve your queries.

Chitosan nanoparticle toxicity: A comprehensive literature review of in vivo and in vitro assessments for medical applications ( )

It could be either. You would need to perform greenhouse experiments to know :)

Using UV disinfection is one of the methods of disinfecting egg cartons.

Bimal Bahadur Kunwar You can try microwave treatment of your soil samples.

Reference:

Brodie, G. (2022). Controlling Weeds with Microwave Energy. In: Horikoshi, S., Brodie, G., Takaki, K., Serpone, N. (eds) Agritech: Innovative Agriculture Using Microwaves and Plasmas. Springer, Singapore. https://doi.org/10.1007/978-981-16-3891-6_8

Ideally you want the seeding to be active and in exponential/log growth phase, so it depends on what stage of the sugarcane fermentation culture is when you use it as seeding: https://lab.plygenind.com/inoculum-preparation-for-fermentation-a-practical-guide

Hello,

Please consider treating your plastics with polyLysine or gelatin. It might help.

P.S.: Some of the cells are definitely degrading over time is stored on -80 instead of -150 or liquid nitrogen tank.

Also many times it is written without any unit means only value is written but yes seed germination, seedling vigour I, and seedling vigour II comes under germination potential of any seed/crop which can be mentioned in percentage. For a better understanding, you can read a few papers and get some idea of how to write the results for SVI and SVII. hereby I have also attached a paper for your kind reference.

You can ask Dr Deepa Agnihotri, Scientist, Birbal sahni Institute of Palaeosciences Lucknow India

available here in Research Gate for this paper.

Ashwini

Hi,

NMR is relatively accurate, or you can purchase a standard sample of canabidiol and compare it using methods such as TLC or GC.

This is feasible. You will need to define and construct your experimental design first, and the particular seed sample you are interested to work with. A mechanical oil presser machine will do to extract the fixed oil. And then, you will need to determine what kind of cell line you want to test it. Probably you would want to investigate its medicinal property thru MTT assay.

The problem may be related to the amount of TPCK-trypsin used in step3 for MDCK cells. The concentration could be too high for the virus inoculation to work properly. A possible solution is to perform a titration experiment to find the optimal concentration of TPCK-trypsin for this procedure.

Paul Reed Hepperly i have overall 900 seeds of each line and first weight of 900 seeds or 100 seeds ?

5 seeds per meter row length

Abiotic nutrients are nutrients in the soil and nothing will be able to grow without them as they are vital. Examples: C, N, P, H, Fe. The abiotic components that help seeds disperse are water and wind, but I also think that temperature may also be involved because the wind more easily blows dry seeds. Light, air, soil, and nutrients, etc. form the abiotic components of an ecosystem. The abiotic factors vary from ecosystem to ecosystem. In an aquatic ecosystem, the abiotic factors may include water pH, sunlight, turbidity, water depth, salinity, available nutrients and dissolved oxygen. The non-living part of the ecosystem is called the abiotic component. This includes physical environments like soil, water, or air. Other includes inorganic compounds, organic compounds or physical factors like temperature. Abiotic components include physical conditions and non-living resources that affect living organisms in terms of growth, maintenance, and reproduction. Resources are distinguished as substances or objects in the environment required by one organism and consumed or otherwise made unavailable for use by other organisms. A terrestrial ecosystem consists of abiotic factors like climate, type of soil or rock, altitude, temperature, nutrients, and minerals, whereas abiotic components in an aquatic ecosystem include dissolved gases, depth of water, salinity, pH of water, light intensity etc.

@ Elias, I guess it should be " Use of plant seed protein for crop production".

I was lacking in the Cellosaurus entry the DT for T84, however I uncovered a paper:

Which has DT for T84 of 33.9 hours in the supplemental table 1. This informaation and the other 52 cell line doubling times reported by this publication will be added to the next release of the Cellosaurus. So your question was very useful, thank you!

Best regards

Amos

Thank you so much

I encountered a similar error in my structural problem with TIE CONSTRAINT. In my case, this error occurred as I permitted the movement of slave nodes to adjust the position initially. Solved this error by unticking the option "Adjust secondary surface initial position" in the interaction section.

@pankaj sharma I tried transfected to cells with 3 plasmids at the same time but it wasnot working. For that reason I am trying to find another solution 😔

Do you have the data for the number of cells? If so, that may be your best option for normalization. The second best option may be to measure protein levels of housekeeping genes and use that data for normalization. Total protein vs your measured hormones will also be better than no normalization at all, but total protein might be affected by your treatment so that would lead to another set of variables.

Hello Innocent Agida

You may please follow the calculations provided below.

You need 5000 cells per well for a 96 well plate.

So, consider 5000 cells for 100 wells (Take extra wells to avoid loss of volume in the final well).

You use the formula:

C1V1=C2V2

Where

C1 = the number of cells that you have (say 2.31 x 10^6 cells /ml)

V1 = volume of original cell suspension to be taken (X ml)

C2 = The total number cells required for 100 wells ( 5000 x 100 = 500 000)

V2 = Total volume for 100 wells (say 100ul per well, therefore 100ul x 100 = 10000ul).

Using the formula

2.31 x 10^6 cell/ml x Xml = 500000 x 10 ml

X = 2.164 ml

You would add 2.16 ml of the original cell suspension (2.31x10^6 cells /ml) to 7.84 ml of culture media to get a total of 500000 cells in 10 ml. Mix well. When you add 100ul of this suspension to each well, you will be adding 5000 cells per well.

Similarly, for 3.09 x 10^6 cell/ml

Use the formula

C1V1=C2V2

C1= the number of cells that you have (3.09 x 10^6 cell/ml)

V1= volume of original cell suspension to be taken (Xml)

C2= The total number cells required for 100 wells ( 5000 x 100 = 500 000)

V2 = Total volume for 100 wells (say 100ul per well, therefore 100 x 100 = 10000ul).

Using the formula

3.09 x 10^6 cell/ml x Xml = 500000 x 10 ml

X = 1.618 ml

You would add 1.61ml of the original cell suspension containing (3.09x10^6 cell/ml) to 8.39ml of culture media to get a total of 500000 cells in 10 ml. Mix well. When you add 100ul of this suspension to each well, you will be adding 5000 cells per well.

Similarly, for 9.65x10^6 cell/ml

Use the formula

C1V1=C2V2

C1= the number of cells that you have (9.65x10^6 cell/ml)

V1= volume of original cell suspension to be taken (Xml)

C2= The total number cells required for 100 wells ( 5000 x 100 = 500 000)

V2 = Total volume for 100 wells (say 100ul per well, therefore 100 x 100 = 10000ul).

Using the formula

9.65x10^6 cell/ml x Xml= 500000 x 10 ml

X = 0.518 ml

You would add 518ul of the original cell suspension containing (9.65x10^6 cell/ml) to 9.482 ml of culture media to get a total of 500000 cells in 10 ml. Mix well. When you add 100ul of this suspension to each well, you will be adding 5000 cells per well.

Finally, for 4.88x10^6 cell/ml

Use the formula

C1V1=C2V2

C1= the number of cells that you have (4.88x10^6 cell/ml)

V1= volume of original cell suspension to be taken (Xml)

C2= The total number cells required for 100 wells ( 5000 x 100 = 500 000)

V2 = Total volume for 100 wells (say 100ul per well, therefore 100 x 100 = 10000ul).

Using the formula

4.88x10^6 cell/ml x Xml = 500000 x 10 ml

X = 1.024 ml

You would add 1.02 ml of the original cell suspension containing (4.88x10^6 cell/ml ) to 8.98 ml of culture media to get a total of 500000 cells in 10 ml. Mix well. When you add 100ul of this suspension to each well, you will be adding 5000 cells per well.

Good Luck!

You are absolutely correct that rainfall before during and after physiological maturity results in fungal infections of seeds which has deleterious effects on the seed quality and its performance.

At the yellow and tan color pod maturity the physiological resistance to seedborne microbes is minimized.

Assuring optimal watering just before flowering and to early pod stages can be sufficient to stimulate yield and quality but moisture on pod surfaces at just before and after physiological maturity will stimulate deleterious seed infections.

You need to dry seeds for its standard moisture and below, then if possible use some fungicide, keep them in a nylon pack, sealed, and freeze-20 C or below. Regards.

Where is the question??

10.1016/j.bcab.2021.102270

Please refer this paper

This will help you

The main thing is that seeds are present :)

The Osmotic Potential of Polyethylene Glycol 6000 (1973)

BURLYN E. MICHEL AND MERRILL R. KAUFMANN

Plant Physiol., 51: 914-916

I have abundoned that field research long ago and cannot help you much.

Prabal Dasgupta

Hello Amanda Marie

It is not okay to transfect cells a week or more after seeding them. The transfection efficiency may decrease. The maximum time frame you can wait to transfect cells after seeding cells is 24 hours.

Cells should be plated 18–24 hours before transfection to ensure that the cells are actively dividing and reach the appropriate cell density (generally 70–80% confluence) at the time of transfection. For higher transfection efficiency, the cells should be > 85% viable at the time of transfection and they should be in early passage.

You should perform a pilot experiment with different seeding densities with the aim of obtaining 70-80% confluency in 24 hours. Then proceed with the transfection experiment using the optimal seeding density.

Best.

GMO means Genetically Modified Organism and specifically modified by genetic engineering where genes from a different animal or plant are put into another. So “non-GMO” seeds just mean that genetic engineering wasn't part of the plant breeding process. GMO means Genetically Modified Organism – and specifically modified by genetic engineering where genes from a different animal or plant are put into another. So “non-GMO” seeds just mean that genetic engineering wasn't part of the plant breeding process. Non-GMO means a product was produced without genetic engineering and its ingredients are not derived from GMOs. Non-GMO Project Verified additionally means that a product is compliant with the Non-GMO Project Standard, which includes stringent provisions for testing, traceability, and segregation. The key difference between GMO and transgenic organism is that GMO is an organism that has an artificially altered genome, while the transgenic organism is a GMO that has an altered genome containing a DNA sequence or gene from a different species. Many GMO crops are used to make ingredients that Americans eat such as cornstarch, corn syrup, corn oil, soybean oil, canola oil, or granulated sugar. A few fresh fruit and vegetables are available in GMO varieties, including potatoes, summer squash, apples, papayas, and pink pineapples. Unnatural as the process of genetically modifying food may seem, GMOs do present a few advantages, compared to organic or conventional / non-GMO products. First of all, they are usually cheaper than their organic or conventional / non-GMO counterparts.

The amount of microbial strain or inoculant needed for seed inoculation depends on several factors, including the type of microorganism, the target crop, and the desired outcome.

In general, a higher inoculum density is required for microorganisms that have a lower survival rate or a lower competitive ability in the soil.

It is also important to consider the dilution effect of the inoculant when it is mixed with the substrate or soil.

The optimal inoculum density for seed inoculation can be determined through field trials and experimentation. It is recommended to follow the manufacturer's instructions for the specific inoculant being used and to adjust the application rate based on the results of previous trials.

Thats what i think .....

Always aim for n = 30 at all levels of replication to fulfil the CLT :)

DAPI is best if the cells are fixed and permeabilized. Label with a very small concentration, just 0.2 ug/mL for 5-10 minutes in PBS or other physiological buffer at RT.

If you wish to do live cell labeling, then I recommend instead using Hoechst 33342, which is more cell permeant, labeled at 0.4 ug/mL for 5 minutes in media or suitable live cell buffer. Be aware that nucleic acids like Hoechst will affect DNA function, such as proliferation, so isn't recommended for culturing or long term imaging after label; only for end point assays.

The surface contamination of the seeds with fungus and being in the right conditions for the growth of the fungus are among the factors that cause the seeds to die

Drought resistance in maize can be evaluated through water stress imposition, measuring physiological traits, root phenotyping, and molecular markers associated with drought resistance. A combination of these approaches can help identify maize varieties that are more tolerant to water stress.

Hi Indranil,

yes you can do it. You can use the cell culture dishes of ibidi. (https://ibidi.com/chambered-coverslips/13--slide-8-well-ibitreat.html or https://ibidi.com/dishes/8--dish-35-mm-high-ibitreat.html?gad=1&gclid=Cj0KCQjwxYOiBhC9ARIsANiEIfZ22ir16PsA0jMbzu6UaC_vkASw9eYa9fJ1TL_8LLEVn7GGNWgH_R8aAsQcEALw_wcB). Hope this helps.

Good luck with your research.

It's a palm tree, which are not cycads, but angiosperms. Cycads are gymnosperms :)

The depth of sowing can have a significant impact on seed germination. Different seeds have different requirements for depth of sowing, depending on their size, shape, and other factors. In general, larger seeds can be sown deeper than smaller seeds, as they have more stored energy and can push through more soil to reach the surface.

If seeds are sown too shallowly, they may dry out or be eaten by birds or other animals before they have a chance to germinate. On the other hand, if seeds are sown too deeply, they may not have enough energy to push through the soil, or may not receive enough light or warmth for germination.

The ideal depth for sowing seeds depends on the specific crop and the conditions in the field. In general, seeds should be sown at a depth that allows them to make good seed-to-soil contact, but not so deep that they are unable to germinate. A general rule of thumb is to sow seeds at a depth that is two to three times their diameter.

It is also important to consider soil moisture when determining the depth of sowing. If the soil is too dry, seeds may need to be sown deeper to reach moisture, while if the soil is too wet, shallow sowing may be more appropriate to prevent the seeds from rotting.

Overall, careful consideration of the depth of sowing can help ensure good germination and healthy crop growth.

There are several reputable sources for domesticated crop seed collection for research purposes. Some of the best sources include:

It is important to note that seed collection for research purposes may require special permits and compliance with regulations. It is recommended to consult with the appropriate authorities before collecting seeds for research purposes.

You should be use a soultion of crystal violet in ammonium oxalat1:10 staining.

A solution:

Crystal violet:2gr

Absolute ethanol 100cc

B solution:

Ammonium oxalat:1gr in 100cc DW

C: 1cc A additive to 9cc DW

1cc C+4cc B

Staining cell cultures after fixed with 7o%ethanol for

Mohammed Fayyadh Some papers about Clavibacter michiganensis:

Hadas, R., et al. "Comparison of extraction procedures and determination of the detection threshold for Clavibacter michiganensis ssp. michiganensis in tomato seeds." Plant Pathology 54.5 (2005): 643-649.

Ansari, Maryam, et al. "Multiple introductions of tomato pathogen Clavibacter michiganensis subsp. michiganensis into Iran as revealed by a global-scale phylogeographic analysis." Applied and Environmental Microbiology 85.24 (2019): e02098-19.

de León, Leandro, et al. "Clavibacter michiganesis subsp. michiganensis, a seedborne tomato pathogen: healthy seeds are still the goal." Plant disease 95.11 (2011): 1328-1338.

Sen, Yusuf, et al. "Bacterial canker of tomato: current knowledge of detection, management, resistance, and interactions." Plant Disease 99.1 (2015): 4-13.

Redo the tests with a larger sample :)

A simple test is the flotation in water test: viable seeds will sink and unsound ones will float :)

What size did you expect? The smaller size of gold NP needs more rpm. A little amount of salt helps the precipitation of gold NP. However, you should be very careful with adding salt to the gold solution. It easily aggregates god NPs. If you provide pictures of your products before and after of the centrifuge, it will help us to find a better solution

Seed development is a complex process involving seed germination (imbibition, emergence of redicle and plumule resulting from bursting of seed due to secretion of GA; and seed emergence...Soil moisture is important whereby seed development may get influenced because of moisture stress (drought) and water logging conditions(devoid of oxygen whereby seed wouldn't be able to respire) because of heavy rainfall or poor drainage...Soil temperatures, humidity, atmospheric temperature are another factor...

Andrew Paul McKenzie Pegman Hi! thanks for replying. I earlier read about mucilage and as I understand mucilage is the collective term used for the secretion by seed during germination. I am looking for specific term for these cells in image. These are single free floating round cells and these are viable even when they are away from the seed coat.

Dear Doraid Sa ,

I'm sorry but you are wrong. You would just need the 10,000 cells and divide that by your actual cell concentration.

10,000/1,025,000=0,0097560 ml or 9.7 µl

You would then take these 9.7 µl and add that to you 80 ml (with such a small volume you do not need to use 79.9903 ml) and after resuspension you can plate them 2 ml per well.

But you should also expect a high mean variation in the cell number per well when you would count them later.

Best wishes

Soenke

Hello, thank you. Interesting to see that this question is stil here. I can also confirm the suggestion. Finally, from the seeds growed Hedera.

Thank you for your help!

Kind regards,

Ute

Thank you very much dear Kouadri,

you've given me a lot of useful and helpful informations.

Please, I want to esk you if you are working on Hedysarum species?

Greetings

Hi Tim,

Have you tried using dielectrophoresis or acoustophoresis to separate the live cells from dead cells/debris? These methods can be cumbersome, but if you're dealing with a small volume of cells they could be OK.

Also check out this paper, they've developed a method for removing non-viable cells and debris from viable cells using inertial microfluidics: https://pubs.rsc.org/en/content/articlelanding/2018/lc/c8lc00250a

Also, it might be worth trying to encapsulate the cells in the ECM material prior to loading your device. If you have a hard time reliably getting Matrigel, it is worth checking out OkaSciences. They make Matrigel equivalents and ship next day! https://okasciences.com/shop/okamatrix/

Hello Ayodeji Abijo

You will have to decide on the seeding density in such a way that the number of cells you decide will help to produce healthy attachment and growth. This number is unique to each cell line, meaning different cell types will have different seeding density requirements based upon their growth characteristics.

Plating cells with different densities can influence the phenotypic characteristics and the growth rate of the cells themselves. If cells are seeded at too low a density, they will go through a lag phase of growth, grow very slowly, or die out completely. If cell densities are allowed to become too high, the cells may exhaust the nutrients in the medium and die abruptly.

The seeding number of 0.8 x 10^6 which you have read for 60mm dish will vary for different cell types depending on their growth characteristics.

Best.

Ruud Nabben no, floating seeds are unviable and the flotation test has been shown to be as reliable as tetrazolium, with the advantage of being non-destructive :)

One simple method for deglycosylation of proteins is using PNGase F, which is an enzyme that cleaves N-linked glycans from glycoproteins. Here is a step-by-step procedure for deglycosylation using PNGase F:

Note: The appropriate amount of PNGase F and reaction time may vary depending on the specific protein and experimental conditions, so it is important to optimize these parameters for each experiment.

This method is relatively simple and can be used for deglycosylation of a variety of glycoproteins, although it may not be suitable for all types of glycans. Other methods for deglycosylation include enzymatic or chemical methods, such as endoglycosidase H or peptide:N-glycosidase F (PNGase A), respectively. The choice of method depends on the specific glycan structure and the desired degree of deglycosylation.

Approx 2-5 x 10^5 cells per well (0.5ml)

Thank you very much Ahmad Al Khraisat

I apologize, I don't know about companies, but maybe you will find them on CANADA or GERMAN platforms.

Please read this review article

Dear David Vicente-Zurdo

I would go with your 2nd option and add the first treatment at a lower volume (100uL). Since the pre-treatment is only for 2 hours I assume that the molecule either binds to something in the surface or enters the cells. In both cases washing with PBS will do very little to remove it. Also, washing cells itself is a treatment which may cause viability loss. Even though you wash all the samples including your non-treated cells, I would avoid extra stress whenever I can. I would only wash if I want to get rid of the vehicle (DMSO, lipofectamine etc.).

By the way, as reduction assays (such as MTT and WST8) cannot differentiate proliferation rate reduction from cell death, they cannot really provide evidence regarding cytotoxicity (only viability loss). I would prefer assays measuring membrane integrity (LDH, PI etc.) instead. Alternatively you can call your measurement "viability change".

For more info on this, I would recommend our article published on plos1:

Cheers

We used to grow bovine aortic endothelial cells and I must say, when I saw your images, they reminded me of some of our cultures. We sometimes observed different morphologies in our cultures that were never fully understood. Maybe when your cells become confluent, they'll appear more endothelial-like? We also observed that endothelial cells appear fibroblast-like when grown on conditioned medium from smooth muscle cells.