VERO Control Culture Transmission Electron Microscopy

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The entire data set, every single image is published here for free download:

VERO CELL CONTROL CULTURE AND TEM IMAGES AND METHOD

Please download and interpret and disseminate your interpretations. This is my interpretation of the opensource results in the way that I see them. Later we will release AI interpretation using numerous different AI programmes, again as a point of reference, but I would encourage anyone else to do the same.

TEM Images

Images were obtained using the equipment listed below:

• Microscope: Tecnai G2 Spirit BioTWIN

• Microscope software: Tecnai version 4.6.4

• Camera: Milexia TEM Camera AMT Nano Sprint 43

• Software: AMT V701 version 7.0.1.525

File naming

Each image file in the download above is named according to this format: TEM_pathX_YYY_Mag_ZZZ.tif

• “X” is the path number, e.g. 1,2.

• “YYY” is an image increment identifier

• “ZZZ” is the magnification, e.g. ’4800’ means 4800x magnification.

TEM METHOD

Culture samples were prepared as follows:

a. Sample fixing:

1. Discard the medium and immediately add 1.5% or 2.5% glutaraldehyde fixative (pH 7.0) to cover the cells.

2. Scrape and collect the cells into a centrifuge tube.

3. Spin down the cells, discard the supernatant.

4. Add fixative mixed 1:1 with medium and gently pellet.

5. Leave at room temperature (RT) for at least 1 hr.

6. Store at 4°C and ship with ice packs.

b. Sample preparation:

1. Cells were fixed for at least 2 hours at RT in the above fixative. They were then washed in 0.1M cacodylate buffer and postfixed with 1% Osmiumtetroxide (OsO4)/1.5% Potassiumferrocyanide (KFeCN6) for 1 hour. This was followed by two washes in water, one wash in Maleate buffer (MB), and incubation in 1% uranylacetate in MB for 1 hour, before two final washes in water and subsequent dehydration through graded alcohol solutions (10 min each; 50%, 70%, 90%, 2x10 min 100%).

2. Samples were then immersed in propylene oxide for 1 hour and infiltrated overnight (ON) in a 1:1 mixture of propylene oxide and TAAB Epon (TAAB Laboratories Equipment Ltd, https://taab.co.uk).

3. The following day, samples were embedded in TAAB Epon and polymerized at 60◦C for 48 hours.

4. Ultrathin sections (approximately 80 nm) were cut using a Reichert UltracutS microtome, collected onto copper grids, and stained with lead citrate.

CONTROL CULTURE COMPARISON

With this TEM session we built upon the first Microscopy Experiments we did where we were only able to have a select few images of one set of reagents. We received just 6 usable images of the 2% FBS (Starvation Medium) Test Cultures. With these new experiments we were able in a 2 hour session, myself and Albert Mathews could work live (Via Zoom) with the Microscopist to not only image the areas of the culture we were interested in, but also to compare and contrast the two paths laid out in the Culture Experiments.

Path 1 was our Negative Control being in 10% FBS, the cells remained healthy and grew to a good confluence over the 5 day incubation period. Path 2, the lowered FBS cultures that replicated Viral Infection Medium showed great amounts of Cell Death and clinical morphology of CPE.

As part of the Preliminary Experiments the CRO Microscopist labelled quite a few cellular features they would like to identify, this was asked as a brief, informal, reference point. All of the labels that the Microscopist chose to label were in Path 1, as it was fairly evident that these cells were much more “Normal/Healthy” in their Biological and Morphological structure.

PATH 1 x890 Mag

PATH 2 x1200 Mag

These greatly zoomed out images of the Vero Cells shows the difference between the Paths from the Light Microscope work in the Culture Experiments. The Cells are elongated and tightly stacked next to each other due to the fact they were scrapped from the culture plates rather than Trypsinized to remove them and prep them for imaging. This decision was made by the CRO to scrape rather than Trypsinize.

The immediate differences on the macro scale in TEM are the large Vacuoles (Holes) present in Path 2 but not in Path 1. This is inline with the CRO noting the large amounts of Vacuoles in the cells which is a supposedly specific Morphological feature of Cytopathic Effect (CPE) at light microscopy zoom levels. As you will come to see these areas of Cell death contained the vast majority of the Nano scale morphological features that are falsely labelled as Viruses. This is also the case with the noted Granular Cytoplasm on the light microscopy level, where larges cytoplasmic structures can be seen in TEM tying the macro and micro scale together to confirm the hypothesis set out for the culture experiments.

There are three areas of these labels that the CRO chose to mark on these Path 1 Vero Cells, they are very important for later on when we interpret the Path 2 images:

MVB: Multi Vesicular Bodies

MVBs are sacks found in every cell that contain Vesicles, round particles that are indistinguishable from Viruses with Microscopy alone. The claim is however, that some should be able to distinguish between these identical particles in infected and uninfected cells.

These Published papers both claim that the only difference between what they label as Clatharin Coated Vesicles/MVBs and Sars Cov 2 is the fact that Sars Cov 2 have quote “Black Dots” inside them and apparently MVBs/CCVs do not (Despite the CCVs in the images definitely having “Black Dots” inside them. They interpret these “Black Dots” are viral proteins in the particles they want to identify as Viral and not in the particles they do not.

(Images are linked to respective publications)

I want to focus on the top Paper as this really shows what they are doing. There is a huge difference between the top image and the bottom in terms of processing. The top image the contrast is whacked right the way up, the lighter areas of the cell line are completely missing they are that white, the darker areas are super sharp. Compare this to the bottom image which is a soup of greyscale where nothing really stands out.

Here I have done this on the MVB that the CRO has labelled in Path 1, the healthy innocuous appearing cell.

Here is the MVB with the soupy GreyScale that the TEM images were given to us in:

Here is the same image with the contrast turned all the way up. Notice those same blurry type vesicles that are claimed not to have the “Black Dots” suddenly appear as soon as the image is sharpened and the debris within the vesicles are shown (Labelled as Viral Proteins for effect):

GLYCOGEN

Glycogen, the sporadic black splodges are dotted around the healthy cells. They are said to be energy storage for the cells. When they are in single or small clusters they lack any sort of clear definition and seem blurred. However in Path 2, in many areas these agglomerated together in large pattern like formations. The Microscopist noted that they didn’t know what this was and had never seen it before. When in this state they blended in with much of the cellular debris and broken down Cellular organelles, again being highly unusual.

Glycogen, being consistently dark stained, relatively Isohedral in shape on their own, yet can be rounded in small clusters and mapping exactly alot of Viruses in size from 30nm singularly up to 200nm in groups. This has led many published papers to retract claims of multiple viruses such as Hepatitis, Sars Cov 2 and Poxviruses as actually mislabeled Glycogen.

FILOPODIA

Filopodia are finger like protrusions that are found in a lot of Animal Cells. The claim is that these protrusions help the cell communicate and be motile and sensory. When these TEM slices are taken, a 2d cross section cannot cut these protrusions in a uniform manner so the result is anywhere from circular where the slide is taken perpendicular to the Filopodia and forms a cross section, or parallel to its longest axis where you would see more of the finger like shape of it.

Filopodia are noted to be in higher quantity in infected Cells particularly with claimed viruses like Sars Cov 2, HIV and Vaccinia. Particularly the cross sections have been misinterpreted as Viruses given their prominent extracellular positions where they claim viruses usually occur, close to the membrane.

FILOPODIAL BRIDGING

Below we see (Paper Linked on Image) a 3D model of how those Filopodia look protruding from the round cells. The claim is, that you can identify a Viral infected cell by a morphology known as a Filopodial Bridge where the protusions between two cells link together.

They Demonstrate in this paper what they claim this Filopodial Bridging looks like in TEM.

Below is two images from our data set, both from Path 2 clearly demonstrating Filopodial Bridging between the separate Cells on the left and right hand side of both images.

Kidney Tubules and Clatharin Coated Vesicles

A Published Peer Reviewed paper looking at kidney biopsies from before 2019 found identical particles to Sars Cov 2 in uninfected and healthy specimens. The paper issued a stark warning saying that due to many particles, in particular Kidney Proximal Tubules could not be distinguished from Sars Cov 2 on their own visually in TEM that a labeling of Sars Cov 2 should not be made on TEM images alone.

Read the full article on that paper below:

Above: The particle, identical in size, shape and claimed corona ring exterior found in the uninfected healthy kidney biopsy. They claim that this particle is a Kidney Proximal Tubule.

Below: Is from a paper entitled “Difficulties in Differentiating Coronaviruses from Subcellular Structures in Human Tissues by Electron Microscopy” where they claim the particle on the right is Sars Cov 2, whereas the two to the left are Clatharin Coated Vesicles. I have highlighted these because they look identical to the claimed Kidney Tubules above. Note the completely ramped up contrast once again to get the interior of the vesicles to pop in black.

Images link to paper

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VIRAL CROSS REFERENCE

In no particular order I will focus in on different Viruses, outline the mainstream description of morphology, characteristics and behavior, showing numerous images all referenced to the publications they are presented in. This will be followed by images of our uninfected cell cultures. In some instances there are exact size measurements that were carried out digitally by the Microscopist. If there is not an exact size carried out, the scale bar will be approximated by measurement as is standard practice in Virological publications.

EBOLA

Mainstream Consensus:

MARBURG

Due to Marburg Virus being morphologically identical to Ebola,these two will be thought of as interchangeable in this falsification of Ebola.

From the Published, Peer Reviewed Journal of Virology images of claimed Ebola Virus in Vero Cells:

Source 2: Claimed Zaire Ebola on Research Gate

Source 3: Claimed Human Cell Expressing Ebola Protein with Ebola Virions visible

Source 4:

Control Culture TEMs:

Interpretation

Demonstrated in the clear images cross referenced that the particles that are being labelled as Ebola Virus are strands of Filopodia, the spacing where they intersect the 2d plain of the TEM slice some cross sectional, some longitudinal showing protrusions is identical in infected and uninfected. The American Society For MicroBiology even uses Vero cells to claim to show Ebola Virions, a cell that is well known to have plenty of Filopodia as shown in our uninfected culture TEM.

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POLIO

(Also Rhinovirus, Coxsackievirus,Echovirus,Enterovirus, Hepatitis A and Parechovirus because they are identical and indestinguishable in TEM)

MAINSTREAM CONSENSUS

Given that there are 7 families of Viruses that are indistinguishable from Polio in TEM I will include references from all of these different families and use them interchangeably to represent this whole class of viruses.

CONTROL CULTURE TEMS

Interpretation

The Microscopist clearly labeled the darker, less defined and encapsulated particles as Glycogen in the healthy cells. In Path 2 these same particles agglomerated into large areas with many hundred all in one place. The Microscopist noted that they had never seen that before. In numerous cross references of Polio, EnteroVirus and Hepatitis they claim these large areas of darker, non distinct particles are a viral infection. They claim ordered particles into rows of three or four are evidence of Progeny where an infection is likely to form. Single Virions that they point to are non distinct and have that characteristic “Rosette” shape attributed to Glycogen.

As shown, glycogen in its singular form and in its agglomeration is being labelled as Polio and 6 or 7 families of viruses.

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MEASLES

MAINSTREAM CONSENSUS

CONTROL CULTURE TEMS

Interpretation

Exact size, shape and inclusion particles to the Measles virus are found within the Path 2 TEMs, singularly and in multiples in the same area. These occur in Vacuoles and areas that have shown cell death where the lipid membrane has broken down and vesicles are forming containing cellular debris. It is clear this breakdown of the cell membrane in vacuoles and the resultant cell debris filled vesicles are being mislabeled as Measles Virus.

Sars Cov 2

Mainstream Consensus

ROUGH ENDOPLASMIC THICKENING

It is claimed that a specific marker of Sars Cov 2 infection of Vero E6 cells is a thickening of the Rough Endoplasmic Reticulum. This develops into cytoplasmic vacuolation.

CONTROL CULTURE TEMS

Interpretation

The mainstream consensus of what the appearance of Sars Cov 2 looks like varies quite drastically. They claim spikes are a prerequisite yet quite a few images do not show spikes, a lot are certainly incomplete. They also claim the interior proteins see as “Black Dots”(Words used in published literature), can tell them apart from MVBs how ever there is no rhyme or reason to the black dots, some are concentric some are spaced evenly some barely visible, all different sizes in comparison to the whole claimed virion.

In our TEMS we found particles of identical size and shape, one or two with visible spikes, all with “black dots”, most found extracellular some with MVBs. It is clear that they are mislabelling MVBs and concentric, rather than more oval with Measles, Cellular Debris encapsulated vesicles.

The TEM samples were stained with a Positive Stain called Lead Citrate, which would render the claimed Spike Proteins invisible. This may explain why those claimed corona are not present in many of the TEM images. Also there is a claim that Trypsin used to remove the cells from the plate to transfer to the TEM slide would create more of these features than the scrapping which was the CRO chosen method.

Because the Viruses Sars Cov I, MERS and Seasonal HCov are absolutely identical in TEM we can incorporate these into this falsification.

HIV

Mainstream Consensus

The mature form of an HIV particle is said to have a conical dense and darker center to it. If taken a cross section through it perpendicular to this conical shape it will show up as varying sizes of sphere. This therefore may be carried over into the falsification as displayed in the first image where all of the particles they label as Mature HIV have spherical dark centers.

CONTROL CULTURE TEMS

Interpretation

The shape of claimed HIV is fairly distinctive outside of the knowledge of Transmission Electron Microscopy. When looking at it however, this type of larger spherical around a smaller spherical darer center is incredibly common. Also located within Path 2 were a few particles with the distinctive conical darkened center. We found inside one vacuole several of these particles in the same area which would be labelled as some sort of infection.

Once again this is vesicular breakdown of the cells into vacuoles, MVBs and Extracellular where darker Cellular debris is encapsulated within the vesicle and mislabeled as HIV. Due to SIV and HTLV being identical in appearance to HIV in TEM we can include those in this falsification.

Rabies

Mainstream Consensus

CONTROL CULTURE TEMS

Interpretation

The claimed clinical diagnosis of Rabies is only done in tissue samples from the brain or brain-stem as finding claimed Rabies virus anywhere else in the body is rare. So TEM of the Vero (African Green Monkey Kidney) should not throw up any cross references. The two defining characteristics of a claimed rabies infection, the Negri Body and the Virion were found in similar areas of Cell Death once again where the membrane has broken down either intercellular or extracellular. The Negri Bodies are electron dense (Darker) areas which suggest they are just part of broken down nucleus being mislabeled.

The distinctive Bullet shape can be found readily across the Cell Line either as mislabeled Endoplasmic Reticulum or Filopodia.

MonkeyPox/SmallPox

The claim once again is that thickening of the Rough Endoplasmic Reticulum is secondary cellular evidence of a MonkeyPox infection. Please refer to the section in Sars Cov 2 on RER thickening to apply across the board. This reference also points to Myelin figures as being a secondary cell morphology of MonkeyPox infection. This will be addressed in the first image below.

CONTROL CULTURE TEMS

Interpretation

The large distinctive blackened virions can be found plentifully in our control TEMS, the lighter in some pictures, darker in others, cylindrical center is also easy to come by and imaged above. The features that go with Cellular Infection such as RER Thickening to vacuolation and Myelin Figures are also present and abundant. Once again cellular debris encapsulated, I would make a suggestion that due to the virion claimed to always be darker than most it is likely cell nucleus that has broken down and the cylindrical centers are part of the Rough Endoplasmic Reticulum as it thickens and breaks down.

Due to SmallPox, CowPox and the rest of the OrthoPox family being identical in TEM we can include these in this falsification of MonkeyPox.

HERPES SIMPLEX VIRUS

With this group of Viruses there are two claimed types the L particle which is considered non infectious but a marker for future infection and the H particle which signifies an active infection.

CONTROL CULTURE TEMS

Interpretation

As with the countless others, these claimed HSV particles are found in areas of cell membrane breakdown whether that be in MVBs, thickened Rough Endoplasmic Reticulum or Vacuoles. This lends itself to the continued notion that these viruses are mislabeled cellular debris encapsulated in cell membrane, The L particle in particular is so so common due to its appearance being identical to just a standard vesicle with no debris filling it.

All of the other identical viruses on TEM appearance to HSV such as Epstein Barre, Varicella Zoster and Cytomegalovirus are to be included in this falsification.

SV40

Mainstream Consensus

The image above is published at this terrible resolution, there is seemingly not a better quality one.

CONTROL CULTURE TEMS

Interpretation

Where there is claimed SV40 in large clusters in the published literature the individual claimed virions lack any sort of definition on close inspection and have an identical dark “rosette” appearance exactly like the Glycogen. The large bodies of claimed SV40 infections are mislabeled glycogen which seems to clump together when the cell is dying.

The individual and budding viruses claimed to be SV40 are hugely common in both pathways, but are most prevalent once again in areas of cell membrane breakdown where encapsulated cellular debris fills vacuoles and thickened Rough Endoplasmic Reticulum and MVBs. These singular claimed SV40 virions are mislabeled encaspulated cellular debris. This falsification also includes JC Virus, BK Virus and Merkel Cell Polyomavirus as they are identical to SV40 in TEM.

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CONCLUSION

In this TEM control culture study we have managed to find exact cross references of the following claimed viruses in uninfected Vero Cells that could not possibly have contained these viruses according to the handling and best practices of the cell banks namely the ATTC:

Ebola

Marburg

Rabies

Sars Cov 2

Sars

Mers

Measles

HIV

HPV

Hep A

Hep B

Hep C

Polio

Herpes Simplex Virus

Varicella Zoster Virus

SIV HTLV1 Human T Lymphocyte Virus 1

Monkeypox

Smallpox

Cytomegalovirus

Epstein-Barr

Human Herpesvirus

Echovirus

EnteroVirus

RhinoVirus

CoxsackieVirus (Lol)

Aichi Virus

Parechovirus

Adenovirus

SV40

JC Virus

BK Virus

Merkel Cell Polyomavirus

We also cross referenced numerous large scale cellular morphologies that are claimed to be indicators of infection as well as specific to infection from claimed specific viruses. These are things such as the increased presence of Filopodia, Filopodial Bridging, Vacuolation, Cytoplasmic Granulation, Rough Endoplasmic Reticulum Thickening and increased size and occurrence off Multi Vesicular Bodies.

The prevalence of these cellular morphologies was not limited to Path 2 as some were seen, such as increased Filopodia in Path 1 but they were certainly predominantly in Path 2. This shows the continuity from the Cell Culture Control Experiments that the conditions of the infection medium alone at reduced FBS concentrations cause Macro and Micro and Nano scale effects of Cytopathic Effect, without the possible presence of a pathogen.

In this study we clearly demonstrate that it is possible to find every single morphology of practically every single pandemic and epidemic causing virus in the Micrographs of uninfected cells. From the long fibrous types of Ebola to the small nucleated HIV to the large repeating patterns of Polio, all of these have identical to the eye counterparts in normal cellular morphology during phases and degrees of cell death.

Given within the published literature the hugely varying morphologies of their own examples of claimed viruses where, with same images labelled as the claimed variety of viruses there was no uniformity to overall shape, encapsulation, colour, matrix within, claimed proteins within, or position in the cell line: There is no real gold standard to their own claimed virological identification in TEM alone.

Demonstrated within this study is the clear cross referencing of all of these morphologies. Due to the sheer volume of these morphologies that we were trying to locate in the 2hr Open Scope session in which the Images were obtained, we were not being specific in our approach, rather looking for overall information.

Bearing in mind that the images of Published and Peer Reviewed viruses are relatively hard to come by, once again with huge variation and often conflicting images between papers. We have no doubt that any criterion that is claimed can identify a virus specifically over and above size, which in all cases were a primary concern in this study, could be satisfied in the same manner and time period by inspecting a control cell line with a specific cellular morphology in mind.

It should be clear by analysing the broad cross spectrum of images in the Published Literature that there is a heavy amount of image editing going on to try and highlight and maybe even alter what they are trying to highlight. This is demonstrated by drastically changing image parameters such as contrast to make certain elements stand out, they have blurred and cropped out or faded away periphery structures that do not fit their hypothesis and also imparted unnatural and simulated colourings once again to give an unnatural highlight to their claimed image.

We chose not to copy this level of editing as it represents in our eyes a level of bias and manipulation to a scientific outcome that is not necessary given the overwhelming weight of our findings.

Further experiments that we would wish to carry out would be to do a comparison between Negative and Positive staining of the same Culture samples to asses the impact of what is visible and what is highlighted. Also to compare and contrast between cells that have been scrapped against cells that have been trypsinized. We know that, as some of the first macro images in this study show (~980x mag) that the scrapping causes the cells to bunch together and flatten out. How that is affecting the morphology, especially of what is seen extracellular with the debris/vesicles/encapsulated vesicles could be speculated to have a fair impact.

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