In this installment of “Testing Greska’s Carbon-60™ for Safety” we will share the results of safety testing conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver and Steep Hill Laboratories in Honolulu, Hawaii.
This week we’d like to share results from testing done on a Transmission Electron Microscope.
The Colorado School of Mines, located in Golden, Colorado, has a TEM and is quite experienced in this type of testing, and has worked with similar materials. We were able to get time to utilize this TEM to review Greska’s Carbon-60 raw powder to clarify the purity and particle size of our product.
PLEASE NOTE: Colorado School of Mines does not endorse Greska’s Carbon-60 product or any other C60 product.
We contracted to use a FEI Talos F200X Transmission Electronic Microscope at an hourly rate to view several samples of raw C60 powder we acquired. This allowed us to observe the professional technician as he operated the TEM to examine and record the properties of these C60 products, including Greska’s Carbon-60 raw powder. All samples (including Greska’s Carbon-60) were presented with no indication of the manufacturer. We also conducted an elemental analysis of the C60 samples.
Before we get to the results, here some background history and additional information about this university where we conducted our testing.
The Transmission Electron Microscope, or TEM, is a FEI Talos F200X CTEM/STEM.
(FEI Co. Talos F200X 200keV field emission scanning / transmission electron microscope)
The FEI Co. Talos F200X 200keV field emission scanning / transmission electron microscope A high total current is conveyed by the FEI X-FEG high brightness electron source, and the integrated EDS system that has four silicon drift detectors (SDDs) allows mapping abilities up to 105 spectra/sec.
This powerful microscope provides a TEM information limit closer than .012nm. A STEM probe provides an imaging resolution equal to 0.16 nm. These STEM images can either be viewed using bright field or using multiple dark field detectors. Also included is a high angle annular dark field detector(HAADF) giving the opportunity for Z-contrast imaging.
TEM Microscope at the Colorado School of Mines
The equipment also features:
So, what does that mean in non-scientific terms? Here’s an explanation in language that us non-scientists can understand:
A Transmission Electron Microscope (TEM) is a very high-powered microscope capable to photomicrograph (a photograph of a microscopic object) pictures with resolution down to the nanoscale level or one billionth of a meter (which is much better than the Micrometer—one Millionth of a meter—resolution limit of a light microscopy).
According to Encyclopedia Britannica, a Transmission Electron Microscope is a “type of electron microscope that has three essential systems: (1) an electron gun, which produces the electron beam, and the condenser system, which focuses the beam onto the object, (2) the image-producing system, consisting of the objective lens, movable specimen stage, and intermediate and projector lenses, which focus the electrons passing through the specimen to form a real, highly magnified image, and (3) the image-recording system, which converts the electron image into some form perceptible to the human eye. The image-recording system usually consists of a fluorescent screen for viewing and focusing the image and a digital camera for permanent records. In addition, a vacuum system, consisting of pumps and their associated gauges and valves, and power supplies are required.”
Transmission electron microscopes are “capable of imaging at a significantly higher resolution than light microscopes, owing to the smaller de Broglie wavelength of electrons,” according to Wikipedia. “This enables the instrument to capture fine detail—even as small as a single column of atoms, which is thousands of times smaller than a resolvable object seen in a light microscope.
“Transmission electron microscopy is a major analytical method in the physical, chemical and biological sciences. TEMs find application in cancer research, virology, and materials science as well as pollution, nanotechnology and semiconductor research.”
Picture courtesy of emresolutions.com
In our application, we used the TEM to perform an elemental analysis using HAADF (High Annular Dark-Field Imaging) on each sample.
We began by taking a very small amount of raw powder—TEM specimens should be less than 100 nanometers thick. This small sample of raw powder was placed on the specimen holder, a very small 3 millimeter (mm) mesh made from copper, with a lacey carbon support film. It is this lacey carbon film that you will see in the comparison pictures below as holes or a grid.
As you can see in the sample picture below, the lacey carbon support film, has many, many tiny holes that vary greatly in size across the span of one single piece of film.
After the raw powder is placed on the copper mesh with the lacey carbon support film, the sample is then placed in the TEM for observation.
The use of electron beams requires that the sample be placed in a vacuum chamber for analysis. An electron beam is produced by applying a high voltage to a hot tungsten filament, and accelerating the emitted electrons through a high electric field, typically 10-100 keV. The electron beam is then focused with magnetic field lenses to a typical spot diameter of 1-100 nm on the sample
Example of lacey carbon film size
When electrons penetrate a sample, they are diffracted to form a diffraction pattern. This diffraction pattern can be transformed with a lens to obtain the sample image. Since the wavelength of an electron is much smaller than the wavelength of visible light, diffraction effects occur at much smaller physical dimensions, enabling us to “see” down to very, very great magnification.
Additionally, a TEM can also detect different types of atoms and the number of these types of atoms in a sample. A simplistic explanation is that an electron beam is introduced which interacts with the atom generating an x-ray whose energy is characteristic of that specific type of atom. The number of x-rays detected as being from carbon atoms, oxygen atoms, silicon atoms, etc., are then used to quantify the amount of each element.
As a reminder, when suppliers or manufacturers claim that their product is 99% or 99.99% pure—they are referring to the ratio of C60 to C70 and higher fullerenes such as C74 and higher. So, when you purchase a gram of 99.9% pure C60 it does not contain 99.9% C60. It contains other “ingredients” or other types of atoms, such as Oxygen, Silicon or Potassium. The TEM microscope has shown a sample of C60 to contain as little as 83% carbon. You can see from the results above, the samples of C60 came back with varying results. Some samples tested fairly pure, containing only carbon and oxygen, including Greska’s Carbon-60. But some samples also tested positive for other elements including, Chlorine, Sulfur, and Magnesium. Some of the samples we tested claim to be vacuum oven baked and some claim to be sublimed.
When we tested Greska’s Carbon-60 raw powder, we found no evidence of higher fullerenes (i.e. fullerenes with 70 carbon atoms or more) only carbon and oxygen. Our product tested as 99.48% carbon and 0.52% oxygen. No other elements were detected. Therefore, in conjunction with the purple test, these tests conclude that Greska’s Carbon-60 is composed of about 99.48% Carbon and about 0.52% oxygen.
At this point, we became curious about the elemental make-up of other carbon 60 products available on the market today. As you know, many manufacturers claim, anywhere from 95% to 99.9% purity, and we wanted to find out what that really means. So, we purchased the highest purity and cleanest C60 raw powder from several other C60 manufacturers to see what their product looked like using the TEM.
We never doubted that these other products contain C60, we were interested in looking at what else was in these products. And what we found was not only interesting, but somewhat alarming for some samples.
In all fairness, we wanted to show all samples at the same magnification, so that we were really comparing “apples to apples.” But as we ran several tests at different times, we did our best to show comparable magnifications. In any case, we believe that a picture is worth a thousand words.
The other very important detail we observed, was the particle size of Greska’s Carbon 60 raw powder was very small. As a comparison, we observed what appeared to be single C60 particles at the 5 nm magnification. As a comparison, we later observed other raw C60 powder samples that appeared as large crystals that ranged from 100 nm to 400 nm in size.
Again, we’d like to mention that you, the reader pay close attention to the particle sizes and the shape of the particles in the pictures below.
IMPORTANT NOTE: Below are pictures of confirmed C60 molecules from third parties. As you will notice when you compare these photos with the pictures taken of Greska’s Carbon-60 with the TEM, you will see that Greska’s Carbon-60 looks remarkably like these photos. Again, another confirmation that our product not only contain C60—but also incredibly small particles of C60! Not only small particles of C60—but the smallest form of C60 commercially available!
Greska’s Sample TEM 120K Magnification
A COMPARISON OF C60 TEM PICTURES:
As we wrap up this article, we’d like to review our previous blogs in order to help our readers understand how all of these pieces fit together to dispel the false implications made in a YouTube video earlier this year.
Other Good Reads
THE TRUTH CONTINUES: SOLUTION VS. SUSPENSION AND HOW THAT AFFECTS GRESKA’S CARBON-60. We shared a video in our blog showing Greska’s Carbon-60, is both a solution and a colloidal suspension. Additionally, because we are both a solution and a colloidal suspension.
There have been many scientific and research studies conducted regarding the toxicity of Carbon 60 and it has been determined that isolated C60 by itself is not toxic. In fact, the Baati rat study published in 2012 was initially intended as a toxicity test. What the scientists found was unexpected! Not only was C60 is not toxic to rats but it appeared to extend the lifespan of the rats that were given C60.
We know Greska’s Carbon-60™ was safe for human consumption, as we are 100% certain our proprietary non-solvent method of producing our C60 uses NO SOLVENTS at any step of the manufacturing process. We then add our C60 to high quality food grade organic sunflower oil. In order to provide the best quality product to our customers, our objective was to verify and provide documentation to our customers that Greska’s Carbon-60™ was not toxic. We wanted confirmation, that not only was Greska’s Carbon-60™ manufacturing process non-toxic but there were no other toxic substances in our product.
Part One of testing of Greska’s Carbon-60™ for toxicity was conducted at the Knoebel Institute for Healthy Aging (KIHA), University of Denver. Located at the base of the Rocky Mountains in Denver, Colorado, KIHA was founded in 2010 when DU received a generous gift from Betty Knoebel. Mrs. Knoebel was the widow of Ferdinand “Fritz” Knoebel, who was a pioneer in the food services industry.
KIHA’s mission is to create and implement solutions for aging issues through multidisciplinary research, education and outreach. Greska’s Carbon-60™ testing was led by Daniel Paredes, Ph.D., Assistant Research Professor whose specialty is aging neurodegenerative disorders such as Parkinson’s, Alzheimer’s and Huntington’s Diseases.
This study was conducted on rat cells, not human cells, therefore we can only confirm that Greska’s Carbon-60™ is non-toxic to rats brain cells.
Samples of neuronal cell culture of rodent (cerebellar granule cells) were exposed to different concentrations of Greska’s Carbon-60™. After 24 hours, there were no signs of any negative effects in the viability, morphology or cell death process as comparted to the non-treated cells.