TEM Image Analysis - Science topic
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Questions related to TEM Image Analysis
I am learning how to edit TEM images in Fiji (Image J).
I want to highlight some specific areas of my images using colors.
Can somebody help?
The TEM image shows the appearance of Moiré fringes with a width of 5-10 nm in the eutectic layer. When strain distribution is analyzed using GPA, strain concentration bands with similar positions to Moiré fringes appear. I would like to know if the presence of such Moire fringes will affect the strain analysis of GPA?
phage tails are getting broken during precipitation and sample preparation, can you suggest a protocol?
The existence of anti-phase domain boundaries (APBs) in polycrystalline materials is usually established by electron microscopic techniques (SEM/TEM)  and is also discussed in diffraction data analyses.
I don’t have a good familiarity with TEM/SEM (and I’m very open to be educated here) but it doesn’t seem convincing enough to look at some microscopic images with atomic level resolution where APBs are found as a straight line (or arbitrarily curved line as in Figure 7 in ref. 1) forming a boundary/wall between the two domains in the same particles, while there is no disorderliness of any sort around and away from the APB.
The reason I’m raising this point is that particle surface is usually more disordered than any kind of defects in the bulk. In fact, it’s even well established that the surface of solid particles behave more or less like a liquid layer , and the smaller the particle size the thicker the liquid layer at the surface. And yet, in the TEM images of nanoparticles that I have seen in some articles there is(are) only the APB(s) visible, and no sign of the bigger unavoidable inherent surface disorder.
Is it possibly due to the fact that in TEM, the electrons pass through the particles and form an image which is influenced by the bulk of the particle? If so, why then the rest of the atomic arrangements within the domains look nearly perfect (i.e. as if it’s a single layer of pointy ordered atoms)?
And as for diffraction data, APBs affect some of the reflections selectively but usually there are different broadening contributions which make it challenging to disentangle. Nevertheless, at least the existence of planar defects like APBs is indicated in diffraction patterns.
Any input would be appreciated.
Article Observation of Surface Melting
One researcher did ballistic performance of kevlar fiber impregnated with nanosilica/ polyethylene glycol shear thickening fluid. And he loaded 10, 15, 20Wt% of Nanosilica with polyethylene glycol.
After fabrication, how we can confirm the equal distribution of nanosilica over the kevlar. Whether nanosilica is spread all over the kevlar equally or not?
How I can confirm? Any SEM, TEM test required ?
For the LFA, detection antibodies need to bound to gold nanoparticles. what are the analytical techniques to verify a successful conjugation? Please suggest any articles with the standard protocol.
I'm having trouble getting clear images of my bacteriophage under TEM. The process I've followed thus far is described below. Any help would be appreciated.
I've isolated bacteriophage active against Staphylococcus aureus bacteria from wastewater.
I grew these to high titer by infecting liquid cultures (S. aureus grown to OD600=0.1) with a low dose of bacteriophage (Multiplicity of infection = 0.1 to 0.05) and incubated until lysis was observed ~3-5 hours later.
These lysates were spun down at 5000g for 5 minutes and filtered through 0.22µm filters.
Chloroform was added (0.1 volumes / 1ml in 10ml), mixed, and left to incubate at room temp for 15-20 minutes with inversions every so often.
These were then centrifuged at 3220g for 10 minutes and care was taken to remove the lysate without touching the cell debris and chloroform layers.
We then used ultrafiltration columns (Cytiva: Vivaspoin 20, 100,000 MWCO) to resuspend the phages (trapped on the column membrane) in pure SM-buffer.
30µl of high titer phage was then fixed using paraformaldehyde and sent for TEM.
TEM images are stained with Uranyl acetate.
I have doped 3%, 4% and 5% of a rare earth metal oxide into a parent material. Is it possible to identify or confirm the dopant through HR-TEM even at these small dopant percentage levels? Also, will the dopants lattice spacing corresponding to the respective hkl plane also be identified?
I have attached two SAED patterns for graphene based nanofillers. Can anyone guide me through the indexing of the said patterns? And also, is it possible to find out the number of graphene layers with the help of diffraction patterns?
Any help will be appreciated.
Assuming that there is a special type of advanced high strength steel that has a fully martensitic microstructure and a nominal tensile strength of 1500 MPa. By changing the composition of a single alloying element, i.e. increasing the C content, we can change the morphology and mechanical properties of the steel. How can we effectively compare the changes between the two steels at a fundamental level that goes beyond the general SEM, EBSD, and TEM characterization methodologies? Are there any crystal plasticity models that can be used in this case? Is a crystal plasticity study even valid for martensitic steels?
Recently I synthesized a material. To specify, I loaded about 0.2%Mg on the synthesized MnO2 nanoparticles, the catalytic performance is very good. I guess that the trace amount of Mg may exist as the single-atom state on the surfaces of MnO2, but I don't know how to chatacterize the Mg. Usually in the publiactions, "heavy" metals such as Pt, Au or Ir acting as single atom catalysts will deposite on "light" supporting materials such as carbon, MOF, Al2O3 or TiO2. And STEM-HAADF will always be used to characterize these single atoms because it can detect and distinguish different elements with big difference in atomic numbers. However, in my research, the single atom Mg (I guess) is lighter than Mn, so STEM-HAADF does not work here. Can anyone give suggestions here? Thanks
Hello, I conducted TEM on cells pre-treated with various compounds, which induced cell stress and death. I originally measured mitochondrial size, swelling of the endoplasmic reticulum, and vacuole presence. However, I read a publication that suggested a difference in vacuole membrane based on type of cell death induced. I re-reviewed my samples, but I'm unsure how to interpret the vacuoles. Some look just like tearing, without a membrane at all.
Any insight or resources appreciated
I used TEM equipped with tungsten gun to obtain attached image (several other too, like shown).
Side blotting technique followed by negative staining (1% PTA for 30 sec) was used. I cannot get the more clear background and there's also problem of resolution.
Please suggest how can I improve sample preparation? or any other suggestion to get better images.
The sample is polyelectrolyte complex in water.
I am working on hollow carbon quantum dots. During the TEM analysis, it is very difficult to get a clearer image of the crystal structure due to the extremely small size. I want to measure the exact size of the shell, hollow-core, etc.
Is there any solution to this problem? I have tested on both: holey carbon grid and the conventional grid but the results are same.
Often research reports will only include only one concentration metric when characterizing nanomaterials. However, often it is necessary to convert among different concentration metrics, does an outline/rubric/methodology exist for how to rigorously perform these conversions?
Two most important structural parameter of a molecule is size and shape. While there is many spectroscopic (like DLS, neutron scattering, fcs) and microscopic (SEM,tem, AFM, etc); for shape measurement as I know, we depend on microscopic techniques.
I want to know other than such imaging techniques are there any such techniques that can measure the shape of nanoparticle or protein?
I heard using XRD it is possible to measure dislocations density or observe them using TEM. I will appreciate if some one explain me how I can measure dislocation density?
I also heard that density of a material can be related to dislocation density, but as I measured the density it was not that much precise to be used in this concept.
Hi everybody. I am trying to analyze some diffractions patterns obtained from TEM, on different parts of the cross-section. The diffraction rings are wide and diffusive because the structure is amorphous but there is still a short-range ordering and I am trying to get more quantitative data from these patterns. So, from one point of the cross-section to another the ring gets wider and more diffusive. I cannot measure its definitive diameter (to calculate d-spacing) since it is diffusive. Any recommendation? most resources and articles are for analysis of crystalline materials.
Dear phage experts,
I recently tried TEM analysis for my isolated phage (H. W. Ackermann, 2011 protocol), but I could not get images and results. Can anyone help me to analyze my phage by TEM?
We are studying organic micelles suspended in water or diesel and want to get images of them to see what they look like (an actual image, rather than size characteristics from something like a DLS). At a minimum, we want to see the micelles after formation, but it would be nice to observe their formation as well. Unfortunately, they are very susceptible to temperature changes so any low-temperature applications likely won't work. I know TEM is an option for this. Is anyone aware of any other options?
Salam all. according to the reported work in the literature, some authors used strong acid (like refluxing with HNo3) in conjugation with carbon-based precursor, while others used base such as NaOH. It is well known that base can act as corrosive agent to accelerate the breakdown process of carbonaceous materials and at the same time provide CDs with OH groups. So what is the advantages of using strong acid? which one is more favorable to enhance the fluorescent quantum yield of CDs , acid or base?
I have synthesized a catalyst for oxygen reduction reaction (ORR). Polyvinylpyrrolidinon was used as as source of C,N while it was pyrolyzed with FeSO4 as a source of Fe and S at 800C in N2 atmosphere using silica template. The EDS shows 0.9% Si, 0.7% S, 0.2% Fe.
Fe may be in the form of Fe3C , Fe2O3, Fe3O4, FeS2 or FeN as per initial XRD observation.
I have seen a lot of TEM images in articles but I could not find similar to my one. Please help me in understanding the TEM images (attached). I have also enclosed SEM images for clear understanding. Thanks in advance!
I want to take the SEM and TEM images of samples I synthesized. I synthesized g-C3N4 (Graphitic Carbon Nitride). If I need to take SEM and TEM, should I disperse the obtained powder in a medium (i.e. water, IPA, etc) or can i do the SEM and TEM tests in powder form?
I've recently imaged a pathogen in the Phylum Ascomycota and am having difficulty discerning between different organelles and possible endosymbiotic bacteria. I can't seem to find any reliable sources showing examples of fungal organelles in TEM images.
I’ve been experiencing difficulties in trying to obtain TEM images of my liposomes. We don’t have a cryo-TEM here in Serbia, so I decided to opt for freeze drying, stabilization using sugars or alditols, different staining techniques etc… Today I got my first images of a newly prepared sample: I freeze dried my liposomal solution (already placed on a grid) over night, and this morning we placed it into a TEM. Any idea why the liposomes (if they’re liposomes) appear like these white shapes inside a big circular dark blurry one? The liposomes are egg phosphatidylcholine + cholesterol + the small organic aromatic molecule I’m encapsulating, in phosphate buffered saline solution. Thank you!
I synthesized AuNP array using protein and DNA. I waned to do element mapping of AuNPs and their surrounding protein and DNA by EDS analysis of TEM equipment. I used protein and DNA to assemble AuNP array, so I wanted to check phosphate, sulfur and nitrogen under the assumption of protein or DNA. However, the peaks of P, S, and N were not detected due to overlapping Au of EDS kev values.(2~2.3kev) Is it really hard to confirm the element of gold particles and DNA or protein together With EDS? I could hardly find paper for element mapping of NP,protein, and DNA together. Anyone know about that issue?? who tried to do it like me?
I have a TEM SAD image and want to determine if the TiO2 is rutile or anatase. I thought I could do this by comparing the d-spacing ratio of different planes. However, I cannot find a database that has the d-spacing between planes.Is there a quick way to calculate, and if so, could someone show me an example for rutile, maybe for the 100 plane or something?
1. The requirement is to do elemental mapping so that we can get compositions of DTS (donor) and PC71BM (acceptor) domains /intermixed regions.
2. We hope to look at the Carbon and the Sulfur Edges by doing EFTEM
3. If we use a Cu TEM grid with the carbon film the signal from the grid might affect the actual signal
so what is the better TEM grid to use ? and if we use a grid with a carbon film is there a possibility to subtract the signal/effect from the film so that we can isolate the actual signal from the active layer ?
I am looking to characterize virus particles within cells via TEM, and am having trouble finding good example images of Retrovirus particles within cells specically type B, C and R. Are there any good references that you could suggest?
Also any other literature or advise on characterization via virus particle morphology, or location within the cell would be helpful as well.
Thank you for your help in advance!
This SAED pattern is generated for a metal semiconductor hybrid nanostructure. I can see it's not single crystalline but I am not sure how to index the planes? Could you please help?
I have been trying to look at blue silver nanoparticles (AgNPs), which are supposed to have a prism shape, on transmission electron microscopy (TEM). However, I have been seeing irregularly-shaped nanoparticles (size ~ 50 - 80 nm) for the majority of time on TEM. I do see some prism shapes but there weren't that many. These AgNPs were prepared fresh and were put on TEM grid within a few days so there should not be degradation due to aging. Also, the UV-Vis spectrum agrees with previously published papers. Just to be sure, I tried putting on AgNPs on the shiny side of the copper grid and also the dull side. I did not see the differences. Could you advise me how you would prepare AgNPs on copper grids ? Thank you!
[Silver Nanoparticles Synthesis (chemical reaction method)]
- Silver nitrate
- Trisodium citrate
- Hydrogen peroxide
- Sodium borohydride
[TEM sample preparation]
- Copper grid with formvar and carbon coated
1) 5 uL of AgNPs were put on the grid
2) Let the grid dry under air in dark
So, I have these four electron diffraction patterns. Can anyone help me for indexing these electron diffraction pattern? Or can anyone give me the step by step to index the electron diffraction pattern?
Thank you in advance.
I have to do the transmission electron microscopy of clay samples. What is a good dispersing agent to be used for the same? In soil engineering as we normally use a solution of sodium hexametaphosphate and sodium carbonate to prevent coagulation of soil, can the same be used for TEM analysis?
I am starting to work with synthesis of silver nanoparticles from natural extracts. I have some pictures of TEM from my samples and I would be grateful if somebody can help me to interpretate them. The difference among them is just the incubation time for the reaction: for the two first photos, it was 1 hour and, for the other two, 24 hours.
Are the darkest spots agglomerates of nanoparticles?
What do you think about the distribution of nanoparticles?
Thank you very much in advance
As you know we have 2 kinds of dislocations in the dislocation accumulation procedure. Statistically Stored Dislocations (SSDs) and Geometrically Necessary Dislocations (GNDs) which either of them can developed by multiplication and strain gradient field, respectively. But in Annealing process which of them can annihilated drastically? in Recovery stage which dislocations can moved easily ?
and is this related to geometrically necessary boundaries (GNBs) and Incidental Dislocation Boundary (IDBs)?
I recently did a TEM on fibroblast cells and noticed that the cells, including control (normal and untreated)cells appear torn (or massive vacoules). The fixing and processing of the cell pellet was performed by a lab that offers these services. I have attached a figure for you to look at. Can anyone tell me if this is normal for skin fibroblast cells or was there an error in the processing? If so, what would you recommend doing?
I use ethanol as a solution and ultrasonicate it for 10-15 minutes and then using a micropippet I pour few drops on carbon coated copper grids and keep it overnight in a oven at 80 deg. But I'm getting an agglomerated micrograph of the sample. And when I'm sintering the sample at a very high temperature, it's fully agglomerated (in microns). So is there any other procedure to get a nicely dispersed particles?
The micro-structure of graphite in SEM is like onion pulp or corn. The main point is how long or how to decide the grain boundaries. The measurement is also very easy. To decide the grain boundaries is difficult task. Any suggestion, I will be very much grateful.
I am size selecting the graphene produced in the lab. While performing Raman analysis, I am finding that the larger flakes have a greater ID/ID' than the smaller flakes. Could anyone please explain why this is happening? (Please see the attached image)
Which aqueous medium is the best for stable and homogeneous DISPERSION of ceramic (PbTiO3) particles for TEM sample preparation ? if particle size is in 500 nm range what precautions must be follow?
I will be appreciated if anyone can help me to know more about the TEM analysis of the core/shell spherical particle. An expert told me that in TEM images (in case of bright field images), when the atomic number and density of a component is higher than the other material, we observe that part darker. But, contrary to his idea, I found some literature as you can find some of them as below which made me confused.
- "A Facile One-Step Approach toward Polymer@SiO2 Core−Shell
Nanoparticles via a Surfactant-Free Miniemulsion Polymerization
Technique": DOI: 10.1021/acs.macromol.6b00038
- "Preparation, characterization and thermal properties of micro-encapsulated phase change materials": https://doi.org/10.1016/j.solmat.2011.09.020
- "Nanometre Ni and core/shell Ni/Au nanoparticles with controllable dimensions synthesized in reverse microemulsion": https://doi.org/10.1016/j.jallcom.2008.07.115
- "Properties of Core−Shell Ni−Au Nanoparticles Synthesized through a Redox-Transmetalation Method in Reverse Microemulsion": DOI: 10.1021/cm070182x
- "Preparation of porous hollow silica spheres via a layer-bylayer
process and the chromatographic performance": DOI 10.1007/s11706-017-0366-z
- "Facile One-Pot Fabrication of Hollow Porous Silica Nanoparticles": DOI: 10.1002/chem.201303656
- " Al2O3/TiO2 core/shell powder derived by novel sol–gel routes": DOI 10.1007/s10971-015-3739-8
- "Increasing the Thermal Storage Capacity of a Phase Change Material by Encapsulation: Preparation and Application in Natural Rubber": DOI: 10.1021/am200870e
- "Nanostructured Materials Through Ultrasonic Spray Pyrolysis": https://www.sigmaaldrich.com/technical-documents/articles/material-matters/ultrasonic-spray-pyrolysis.html
So, I am wondering if there are any other parameters that can be effective in TEM analysis of core/shell structure particles along with the atomic number and density of the material? I will be appreciated If you can also introduce me some references about TEM analysis of core/shell spherical particles.
Thank you so much in advance.
Interfaces in composites are expected to be non-planar. What attempts have been made to provide expressions for the displacement and stress fields due to dislocations and cracks that lie on a non-planar interface in three dimensions?
I am trying to get information about stacking fault and dislocation based on TEM image. In the attached figure, I process the image using FFT implemented in digital micrograph DM. Are the circled locations are evidence for SF and dislocation. I really will learn a lot from your comments.
Thanks in advance
when i want to try thick sectioning of my sample, I fill in the tray with water bu even with angular and axial regulations and also filling and vacuuming water , it does not wet the upper part ! and so does not approach the edge ... I do not know what is the problem...is it from the water or the glass or maybe cleanliness.... ?please advice
How does the TEM electron beam change when I key the dark field button on the panel?
And I do not understand the meaning of "Use Multifunction X and Y knobs to bring the Bragg reflection opposite to the one selected to the point" in the axial dark-field imaging manual.
Why move the DP twice?
Attachment is the Tecnai F20/F30 Operating Manual axial dark-field imaging operating manual.
how i can calculate Lattice parameter from TEM image using software Gatan. i want to measure the matrix lattice and the precipitate
I confronted some difficulty in understanding the convergence semi-angle on the condition of over-focus.
The convergence angle in the condition of focus like that Fig.1 shows. however, the over-focus condition seems different, as Fig.2 shows.
The defination of canvergence angle under focus/overfocus is the same? How can we explain or understand that?
I had a 3D model to study myelination in vitro. having TEM pictures of my cells being myelinated by schwann cells, I wanted to analyse my pictures, but I don't know how.
I'll be glad if you guys tell me so.
When i took the TEM images of CuS-Ag hybrid nanoparticles, i noticed that some nanoparticles have a vibrative bubble inside.
Does anyone can tell me why this happen?
This nanoparticle synthesized in aqueous manner.
We are planning to do a pre-embedding immunoreaction for EM. Although there are many alternatives we will try the DAB reaction on HRP-conjugated Ab, followed by silver precipitation (Gallyas, 1982) and gold substitution: GSSP-protocol.
One of the key chemicals of the GSSP-procedure is gold chloride (AuCl3). We don’t have that anymore. What we do have is the Nanoprobe GoldEnhance for EM plus (GEEM+). Do you know if this GEEM-solution can be used to replace the silver-precipitate?
Thanks for your time
From my understanding, I need to focus the beam on sample by reduce the intensity. Then, click diffraction mode, and then increases intensity knob. For some reason, I still don't see any kikuchi line. Anyone tell me if my operation of TEM is right?
At the bottom of the SAED image, there is a scale marked 2 1/nm, can someone just clarify what is it, why 2 1/nm marked, and how it is used while calculation??? I have attached the reference image.
I am working on a crystal, and collected both electron diffraction pattern (ED) and XRD pattern. The strange thing is, one ring (low index) in ED can not be assigned to any peaks in XRD. How does this happen? is there any difference in diffraction by using electron beam or x-ray?
I was given some liver tissue samples for viewing using scanning and transmission electron microscopes. The samples were kept in McDowell's solution at 4 degree Celcius for several months up to a year.
I wondered if the samples were already over-fixated by this time?
I´d like to ask, what can be an origin of the dark (electron-dense) objects (whole first picture) and in the second picture (chloroplast), it is present in the very left side, right side and above chloroplast. These dark objects are appearing during contrasting the sections (uranylacetate and lead citrate). Very often it covers all structures in the cells/tissueas and it is impossible to obtain good photos. We observed also non-contrasted samples and in this case there are no dark objects. I´ve tried also different concentrations od lead citrate (0.2 - 2 %; pH 12-13), but the objects are still present. I use 2% uranylacetate (pH cca 4) (contrasting for 35 min.), then 6x washing by MiliQ water (room temperature, afted boiling, to avoid CO2) and then contrasting by lead citrate for 2.5 min. We are sure these disturbing objects are appearing during contrasting process, but we don´t know how to avoid them. Thanks.
I exported my EBSD data (acquired using TSL OIM Data Collection S/W) in .ang format and saved it as a separate file from the .osc file. However, reopening the .ang file in TSL OIM Analysis gives the error: "Unable to open the file: Invalid symmetry". The .osc file works smoothly though.
Why is it so? And the same .ang files also don't work in MTEX where I need to post-process my data for many important results.