Figure 1 – CryoTEM images of adenovirus samples the classification on the right was performed having in account the internal density profile of each particle.
Figure 2 - Graphical representation of the high density (Type I) versus low density (Type II) analysis. CryoTEM image, the detection of adenovirus particles is overlay in the image with red and blue circles.
Figure 3 - nsTEM image of an adenovirus sample. The visualization of the disassembly of adenovirus segments, hexons and fibers (left imaged) as the presence of impurities in the samples, will be helping in the choice of the right purification method in the lab.
Figure 4 - Graphical representation of the individual versus cluster/aggregates analysis. In the nsTEM image (right), the detected cluster/aggregates and the individual adenovirus particles are overlayered with red and blue circles, respectively.
Figure 5 - CryoTEM (upper) and nsTEM (bottom) images of adenovirus samples. The image on the right has the classification overlaid having in account the integrity of the adenovirus particles (green circles intact particles and yellow circles broken particles).
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Due to technical differences nsTEM is more suitable for aggregation studies. Clustering/aggregation analysis is recommended to be used for comparison studies between samples having similar formulation, to obtain a relative assessment of their cluster/aggregate state. The analysis of an individual sample is also possible although this needs to be optimized by us for your own product. Please contact us for further information.
| Reagent type | Reagent name | Concentration range | Impact in the sample preparation. |
| Buffer component | Phosphate | Minimum possible concentration |
Cause interaction with nsTEM resulting in crystalline precipitates. |
| Sugars | Glucose, trehalose, glycerol and others | 0-10 % | Influences the contrast inside the microscope difficulting the imaging and preparation. Usually high concentrations of sugars will lead to unspreading of the samples on the grid. |
| Detergents | Triton X-100, SDS and others | 0-10 % Dependent on the detergent cmc |
Affects both stain and ice formation in nsTEM and cryoTEM respectively. |
Reducing agents |
Ascorbic acid, 2-mercaptoethanol. DTT and others | Minimum possible concentration | Interaction with the stain used in the nsTEM resulting in precipitates. |
| Salts | NaCl and others | 0-300 mM | May induce salt crystallization in nsTEM. In cryoTEM the presence of salt may induce to a noisy background that prevents contrast in the microscope. |
| Grow media | Any enriched growth media | Any |
Prevents the particle on grid distribution and due to high protein concentration will create a noisy background in the images. |
Yes, we do provide non-GMP analysis in case there is no need for GMP analysis.
We can perform TEM analysis for samples up to biosafety level 2 at our GMP certified laboratory in Stockholm.
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