Transmission Electron Microscope (TEM) Imaging Service

Introduction

Using the advanced techniques of Transmission Electron Microscopy (TEM) and its counterpart, Scanning Transmission Electron Microscopy (STEM), high-speed electrons pass through ultra-thin specimens that are electron-permeable (around 100 nm thick). As these electrons traverse the specimen, interactions occur, resulting in the genesis of detailed imagery. This imagery is further refined in the case of STEM, by concentrating the electron beam to a minute point and methodically scanning it across the sample in a grid pattern. Such techniques engender high-fidelity atomic scale visualizations, with resolutions in the range of 1-2Å. These meticulous visual representations have found profound applications across diverse fields, incorporating oncology research, virology, materials science, and other foundational research disciplines.

Fig.1 TEM image of an ultrathin section of a plastic-embedded liver sinusoidal endothelial cell after isolation¹.

Sample Requirements

• Animal and Plant Tissue
  1. Swift action: Once the tissue is isolated, swiftly submerge it into electron microscope fixation solution so the tissue cells maintain their original state as much as possible.
  2. Minimize damage: Use sharp cutting tools to reduce the pulling or squeezing of the tissue.
  3. Tissue size: The tissue sample should be about the size of a grain of rice and as thin as possible.
  4. Sampling location: Aim to acquire the target location; clumping or rolling animal tissue samples can be adhered to filter paper for fixation. For less dense plant tissue samples, add appropriate objects to ensure ample contact between the fixative and tissue.
  5. Transport conditions: After fresh samples are fixed at room temperature, transfer them to 4°C for storage, and transport them in a foam box with ice packs to the lab. Avoid direct contact with the ice pack to prevent the fixative from freezing.
• Cells
  1. Try to use a 60mm dish for cell culture, collect in a centrifuge tube, centrifuge at 1500 rpm for 10 minutes, and discard the supernatant.
  2. Slowly add approximately 0.5% glutaraldehyde fixative solution (diluted 1:6 with PBS) along the wall of the tube, and let it sit for 10 minutes at 4°C.
  3. Then centrifuge at 10000-13000 rpm for 10-15 minutes and discard the supernatant.
  4. Slowly add 3% glutaraldehyde fixative solution along the wall of the tube. If the sample cannot be sent immediately after collection, it can be stored at 4°C. The volume of the sample after high-speed centrifugation should not be less than half the size of a green bean.
  5. Transport conditions: After centrifuging and discarding the fixative, add new electron microscope fixative, transfer to 4°C for storage, and transport in a foam box with ice packs to the lab. Avoid direct contact with the ice pack to prevent the fixative from freezing.
• Bacteria
  1. Bacterial collection: Directly scoop up the visible bacterial colonies on the solid culture medium and place them in the fixative along with the culture medium. Suspended bacteria and others need to be collected by centrifugal fixation.
  2. Transport conditions: After fixation at room temperature, transfer to 4°C for storage, and transport in a foam box with ice packs to the lab. Avoid direct contact with the ice pack to prevent the fixative from freezing.
• Others
  1. Nanomaterials: Can prepare suspensions or powders, don't use too little sample.
  2. Others: For tissues or specimens that are difficult to collect or sample, please call for consultation.
  3. Transport conditions: After fixation at room temperature, transfer to 4°C for storage, and transport in a foam box with ice packs to the lab. Avoid direct contact with the ice pack to prevent the fixative from freezing.
• Precautions:
  1. Transmit samples at 4°C to the lab. The recommended volume ratio of fixative to tissue samples is 10:1.
  2. For cell (bacterial) samples, use a pointed bottom EP tube of about 1.5mL, centrifuge into clusters (about the size of a grain of rice), the sample should not disperse.
  3. The sample should not be too large, and the fixative should not be too little.
  4. Only choose 3% (or 2.5%) glutaraldehyde as the fixative.

Service Turnaround Time and Deliverables

Service Turnaround Time:

The experiment is expected to be completed within approximately 2 weeks.

Deliverables:

The deliverables will include electron microscope images, pathology reports, and experimental reports detailing the experimental procedures, reagents, and instruments used.

Creative Biolabs' TEM Service

Creative Biolabs delivers a rapid, adaptable, and dependable TEM (Transmission Electron Microscopy) service, ensuring results within a mere 15 days. Specializing in intricate structural specimens, this service addresses diverse biological and inorganic samples, from organs to creatures and nanoparticles, extending to viruses, vesicles, and bacteria. Rigorous guidelines paired with unwavering quality assurance make it an invaluable choice for researchers. Partnering with Creative Biolabs ensures top-tier support for your TEM imaging requirements.

For more information, please contact us.

FAQs

Q1: What types of samples can be analyzed using Creative Biolabs' TEM service?

A: Creative Biolabs' TEM service can analyze a wide variety of samples, including biological tissues, cells, inorganic materials, and more.

Q2: How does Creative Biolabs ensure the quality of the TEM imaging?

A: Creative Biolabs provides detailed guidelines for sample preparation and follows a rigorous process to ensure the quality of the TEM imaging.

Q3: How to prepare a 2.5% glutaraldehyde fixative?

• Commercially available 25% glutaraldehyde aqueous solution: 10 ml
• 0.2 M phosphate buffer (pH7.0): 50ml
• Dilute to 100 ml with distilled water.

Q4: How to prepare a 0.2 M phosphate buffer with pH 7.0?

• Solution A: Dissolve 31.61 g of Na₂HPO₄·H₂O, or 53.63 g of Na₂HPO₄·7H₂O, or 71.64 g of Na₂HPO₄·12H₂O in distilled water to make up to 1000 ml.
• Solution B: Dissolve 27.6 g of NaH₂PO₄·H₂O, or 31.21 g of NaH₂PO₄·2H₂O in distilled water to make up to 1000 ml.
• The 0.2 M phosphate buffer (pH 7.0) is made by combining 61 ml of Solution A with 39 ml of Solution B (to make a total of 100 ml).

Reference

1. Wisse, Eddie, et al. "Fixation methods for electron microscopy of human and other liver." World journal of gastroenterology: WJG 16.23 (2010): 2851.