Unlike most cores and commercial providers, we customize our TEM services for each client’s science. We have a wide range of variations to customize our electron microscopy services and costs are based on actual staff efforts and consumables.
Costs Are Per Project, Not Per Specimen
If multiple specimens can be incubated and batch processed in parallel, we will pass the savings directly back to the user. Solo specimens are the most expensive, but each additional specimen reduces the unit costs. The maximum number of specimens that can be batch processed depends on the complexity of the processing required by the scientific goals.
Note: Estimates based on FY2019 JHU-academic rates. |
||||||||||||||||||||||||
Range of TEM Services and Project Customization
- Histology of ‘semi-thin’ sections prior to thin-sectioning allows us to locate the interesting regions of your specimen with your guidance. Remember, ultra-thin sections used in TEM are only ~90 nm thick. Typically, we use toluidine blue staining of ‘semi-thin’ sections (~500 nm thick). We will use the histology images to consult with you about regions of interest in the block. The process is iterative, so takes more work and more time; see itemized costs of histology below.
- Immuno-Electron Microscopy reveals the location of molecular epitopes using small gold particles (~5 nm) and antibody labelling in TEM specimens. However, standard TEM processing can bury or destroy antigenicity. We might need to perform antigen-recovery, which chemically etches the Epon epoxy after ultra-thin sectioning, revealing more epitopes for antibody binding. Another alternative is to use hydrophilic acrylic resins (e.g. LR White), rather than the standard Epon epoxy resin. Prior literature and experience will guide choices. Because varying concentrations of antibody and related reagents must be explored, we limit projects to two specimens (including negative control) for immuno-EM studies. As examples, see immuno-EM costs itemized.
- High-Pressure Freezing (HPF) can avoid artifacts of fixation and permeability, as well as providing exquisite membrane details with automated freeze-substitution (AFS). Specimens shown to benefit from high-pressure freezing include nematodes (C. elegans), fruit fly embryos (D. melanogaster), and yeast (S. cerevisiae). As examples, see HPF/AFS costs itemized.
- Microwave Fixation/Staining has proven invaluable for penetrating dense tissues and for visualizing myelination layers. Temperatures are carefully controlled to prevent bulk heating, but reagents penetrate better with microwave processing. The additional costs are negligible.
- Share the Labor of TEM projects. Unlike full-service projects, where we do everything after you drop off your specimens, you can choose to do some of the labor in the project, thus saving in the costs. For example, if you’re trained to operate the electron microscope, you can view the prepared grids on your own completely without us. Of course, training costs extra and takes time to become as efficient as our staff. Another way to share the labor is to perform the ultra-thin sectioning yourself, assuming you have been trained to use the ultra-microtomes previously. These two examples of sharing the labor are itemized below, but we can share the labor in other ways. Contact us to explore the possibilities.
Advanced EM Techniques with Their Approximate Costs
For new immuno-EM projects, we must explore multiple labeling conditions, varying the concentrations of both antibody and secondary reagents, and the conditions are often grouped into two separate attempts, each with fresh resin-embedded sections. If the first attempt works, then the second attempt is unnecessary. In addition, we limit immuno-EM projects to two specimens to keep the number of variations manageable. Studies typically double the number of grids to allow for Ab-staining controls (lacking primary antibody).
| Customization | Add’l Cost | Total Cost, Full-service 2 specimens |
Add’l Timing | Total Timing (typical) |
|---|---|---|---|---|
| Immuno-Electron Microscopy (LR-White resin, single user-provided antibody) |
add ~$715/Ab/Attempt | $1,625- $2,470 | add 2-3d per Attempt | 18 days |
| Antigen-recovery, chemical etching Epon-epoxy after sectioning (can recover antigenicity) | add ~$845/Ab/Attempt | $1,860- $2,730 | add 2-3d per Attempt | 18 days |
| High-Pressure Freezing (HPF) & Freeze-Substitution (AFS) |
add $305/Proj | $1,430 | add 7 days | 21 days |
| Immuno-EM of HPF/AFS-prepared Specimens | add ~$715/Ab/Attempt | $1,930- $2,775 | add 2-3d per Attempt | 27 days |
Note: Estimates based on FY2019 JHU-academic rates.
Examples of Project Itemization in Statements
Billing statements for electron microscopy projects will list the staff person’s effort on the project, broken into activity categories. Below are a few examples to show how some customizations are itemized and how they compare in pricing. Remember, your specimens and your scientific goals may introduce other considerations that affect the final costs and differ from these example itemizations.
Itemization: Full-service, 2 vs 4 specimens, No histology
These full-service estimates assume tissue specimens provided as ~1mm3, that no histological staining is needed to identify regions for ultra-thin sectioning, and that you will sit at our side, guiding while we operate the microscope. Different specimen sources (e.g. cell culture vs tissue) will affect the estimate ~0.5h/specimen. Your scientific goals may introduce other considerations that affect the final costs.
| Description | 2 specimens | 4 specimens | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.3h | $130/h | $690.00 | 5.6h | $130/h | $730.00 |
| TEM Sectioning (~0.5h/specimen) | 1.0h | $130/h | $130.00 | 2.0h | $130/h | $260.00 |
| TEM Staining (2 grids/specimen) | 1.0h | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| Microscopy, Assisted Use (~0.5h/specimen) | 1.0h | $130/h | $130.00 | 2.0h | $130/h | $260.00 |
| Total Cost for Project | $1,080.00 | $1,380.00 | ||||
Note: Sectioning and staining estimates assume 2 grids/specimen. Estimates based on FY2019 JHU-academic rates.
Itemization: Cost of Histology, Full-service, 2 specimens
These full-service estimates compare the extra work required for histological staining to identify regions for subsequent ultra-thin sectioning. It may take multiple iterations that will affect costs and add delays while we wait for feedback from clients. Your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | No Histology | Add’l Histology | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.3h | $130/h | $690.00 | 5.3h | $130/h | $690.00 |
| TEM Sectioning (~0.5 vs ~1h/specimen) | 1.0h | $130/h | $130.00 | 2.0h | $130/h | $260.00 |
| TEM Staining (2 grids/specimen) | 1.0h | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| Microscopy, Assisted Use (~0.5h/specimen) | 1.0h | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| Total Cost for Project | $1,080.00 | $1,210.00 | ||||
Note: Sectioning and staining estimates assume 2 grids/specimen. Estimates based on FY2019 JHU-academic rates.
Itemization: Full-service vs Self-Microscopy, 4 specimens
To operate the microscope yourself is much cheaper ($29/h), but requires training (typically 3h, ~$350) and practice to become proficient. If you expect to need a lot of electron microscopy, investing in training and practice makes sense and saves a lot of money in the long-run. We also provide ‘semi-assisted’ use of the microscope: we’ll set up and focus the microscope for each specimen, but we’ll leave you to operate the microscope to choose the different fields of view to acquire. Semi-Assisted Use ($65/h) costs half the rate of Assisted Use and must be arranged in advance to ensure staff availability. As always, your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | Full-Service | Self-Microscopy | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.6h | $130/h | $730.00 | 5.6h | $130/h | $730.00 |
| TEM Sectioning (~0.5h/specimen) | 2.0h | $130/h | $260.00 | 2.0h | $130/h | $260.00 |
| TEM Staining (2 grids/specimen) | 1.0h | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| Microscopy, Assisted Use (~0.5h/specimen) | 2.0h | $130/h | $260.00 | |||
| Microscopy, Self-Use (~0.5h/specimen) | 2.0h | $29/h | $58.00 | |||
| Total Cost for Project | $1,380.00 | $1,178.00 | ||||
Note: Sectioning and staining estimates assume 2 grids/specimen. Estimates based on FY2019 JHU-academic rates.
Itemization: Full-service vs Self: Sectioning, Staining & Microscopy, 4 specimens
The more labor you provide, the less your costs. In addition to operating the microscope yourself, you could perform the histology and ultra-microtome sectioning yourself. To operate the ultra-microtome requires training (typically 2h, ~$260) and practice to become proficient. In addition, you would need to invest in your own diamond knife should glass knives prove insufficient. Considerations for operating the microscope yourself were described above. As always, your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | Full-Service | Self: Section, Stain & Microscopy | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.6h | $130/h | $730.00 | 5.6h | $130/h | $730.00 |
| TEM Sectioning (~0.5h/specimen) | 2.0h | $130/h | $260.00 | |||
| Ultra-Microtome, Self-Use (~0.5h/specimen) | 2.0h | $16/h | $32.00 | |||
| TEM Staining (2 grids/specimen) | 1.0h | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| Microscopy, Assisted Use (~0.5h/specimen) | 2.0h | $130/h | $260.00 | |||
| Microscopy, Self-Use (~0.5h/specimen) | 2.0h | $29/h | $58.00 | |||
| Total Cost for Project | $1,380.00 | $845.00 | ||||
Note: Sectioning and staining estimates assume 2 grids/specimen. Estimates based on FY2019 JHU-academic rates.
Itemization: Full-service, Std (Epon) vs Immuno-EM (LR-White), 2 specimens
For immuno-EM, extra labor is required to explore the different concentrations of antibodies and related reagents. Hence, we limit immuno-EM to only 2 specimens (include negative control). We make no more than two attempts at post-sectioned antibody-labeling, including controls (lacking primary antibody), using a range of reagent concentrations. If the first attempt succeeds, the second attempt is not necessary and we pass the savings back to you. Because of the extra grids with differing reagent concentrations, microscope imaging time is necessarily longer than non-immuno-EM studies. As always, your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | Full-Service (not immuno) | Immuno-EM (LR-White) | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.5h | $130/h | $715.00 | 5.5h | $130/h | $715.00 |
| TEM Sectioning (~0.75h/specimen) | 1.5h | $130/h | $195.00 | 1.5h | $130/h | $195.00 |
| TEM Staining (includes Ab labeling) | 0.65h | $130/h | $85.00 | 3.5h | $130/h | $455.00 |
| Microscopy, Assisted Use | 1.0h | $130/h | $130.00 | 2.0h | $130/h | $260.00 |
| 2nd TEM Sectioning (~0.5h/specimen) | 1.0h | $130/h | $130.00 | |||
| 2nd TEM Staining (Ab labeling) |
3.5h | $130/h | $455.00 | |||
| 2nd Microscopy, Assisted Use | 2.0h | $130/h | $260.00 | |||
| Total Cost for Project | $1,125.00 | $1,625.00- $2,470.00 | ||||
Note: Estimates based on FY2019 JHU-academic rates.
Itemization: Immuno-EM: LR-White vs Antigen-retrieval, 2 specimens
Because standard Epon-epoxy TEM processing can bury or destroy antigenicity, techniques have been developed to preserve antibody labeling. Only empirical testing can determine which technique is best. Two popular techniques are using hydrophilic acrylic resin (e.g. LR-White) or to chemically etch Epon-epoxy (‘antigen-retrieval’). Here, we compare the itemization of these two techniques, including the possibility of needing only one attempt at antibody labeling. As always, your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | Immuno-EM (LR-White) | Immuno-EM (Ag-Retrieval) | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| TEM Projects/Techniques (fixation, resin embedding) |
5.5h | $130/h | $715.00 | 5.5h | $130/h | $715.00 |
| TEM Sectioning (~0.75h/specimen) | 1.5h | $130/h | $195.00 | 1.5h | $130/h | $195.00 |
| TEM Projects/Techniques (antigen-retrieval) | 1.0h | $130/h | $130.00 | |||
| TEM Staining (includes Ab labeling) | 3.5h | $130/h | $455.00 | 3.5h | $130/h | $455.00 |
| Microscopy, Assisted Use | 2.0h | $130/h | $260.00 | 2.0h | $130/h | $260.00 |
| 2nd TEM Sectioning (~0.5h/specimen) | 1.oh | $130/h | $130.00 | 1.0h | $130/h | $130.00 |
| 2nd TEM Projects/Techniques (2nd antigen-retrieval) | 1.0h | $130/h | $130.00 | |||
| 2nd TEM Staining (second Ab labeling) |
3.5h | $130/h | $455.00 | 3.5h | $130/h | $455.00 |
| 2nd Microscopy, Assisted Use | 2.0h | $130/h | $260.00 | 2.0h | $130/h | $260.00 |
| Total Cost for Project | $1,625.00- $2,470.00 | $1,755.00- $2,730.00 | ||||
Note: Estimates based on FY2019 JHU-academic rates.
Itemization: High-Pressure Freezing, Std (Epon) vs Immuno-EM (LR-White), 2 specimens
High-pressure freezing and freeze-substitution are great ways to avoid fixation or permeabilization artifacts, often enhancing the resolution of staining (each leaflet of membrane bilayers are resolvable!). It can be combined with immuno-techniques, too. Below is an example of the itemization for each type of HPF/AFS projects. As always, your specimen and scientific goals may introduce other considerations that affect the final costs.
| Description | HPF/AFS (not immuno) | HPF/AFS Immuno-EM (LR-White) | ||||
|---|---|---|---|---|---|---|
| Hrs | $/h | Cost | Hrs | $/h | Cost | |
| HPF Pre-chill (liquid nitrogen) | 1 | $200/sess | $200.00 | 1 | $200/sess | $200.00 |
| TEM Projects/Techniques (HPF freezing, ~0.75h/spec) |
1.5h | $130/h | $195.00 | 1.5h | $130/h | $195.00 |
| HPF Freezing Supplies (triplicate per spec) |
6 | $10/freeze | $60.00 | 6 | $10/freeze | $60.00 |
| AFS Pre-chill (liquid nitrogen) |
1 | $110/sess | $110.00 | 1 | $110/sess | $110.00 |
| TEM Projects/Techniques (fixation, resin embedding) |
5.6h | $130/h | $730.00 | 5.6h | $130/h | $730.00 |
| TEM Sectioning (~0.75h/specimen) | 1.5h | $130/h | $195.00 | 1.5h | $130/h | $195.00 |
| TEM Staining (includes Ab labeling) | 0.65h | $130/h | $85.00 | 3.5h | $130/h | $455.00 |
| Microscopy, Assisted Use | 1.0h | $130/h | $130.00 | 2.0h | $130/h | $260.00 |
| 2nd TEM Sectioning (~0.5h/specimen) | 1.0h | $130/h | $130.00 | |||
| 2nd TEM Staining (Ab labeling) |
3.5h | $130/h | $455.00 | |||
| 2nd Microscopy, Assisted Use | 2.0h | $130/h | $260.00 | |||
| Total Cost for Project | $1,430.00 | $1,930.00- $2,775.00 | ||||
Note: Estimates based on FY2019 JHU-academic rates.