Electron Microscopy Facility 

Frequently Asked Questions

General Questions
Clinical Application Questions
   Platelets and Storage Pool Deficiency
   Muscle Biopsies
   Nerve Biopsies
   Renal Biopsies
   Tumors
   Bone Marrow
Research Questions
Atlas Questions


General Questions
Why use a transmission electron microscope (TEM)?
Do you have a scanning electron microscope (SEM)?
How much tissue is needed for study?
Why does the sample have to be so small?
What advantages does the TEM offer over light and/or fluorescence microscopy?
How long does it take to process tissue?
What types of TEM procedures are available?
Why can’t frozen samples be used for TEM?
Why are TEM images in black and white/grey scale?
I still have questions….

Clinical Application Questions

What clinical electron microscopy services do you offer?
How should I send my sample?
Can I send a sample at any time?
Should I chill or freeze my sample?
How will you bill my sample?
When should I expect a report?
If I haven’t received a report, who should I contact?
    Platelets and Storage Pool Deficiency
What is delta (dense) granule storage pool deficiency?
How many delta granules should platelets have?
What are common symptoms of delta granule storage pool deficiency?
What is the CPT code for a Platelet Storage Pool Deficiency Analysis?
    Muscle Biopsies
How should we prepare and ship a muscle biopsy specimen for an EM study?
    Nerve Biopsies

How should we prepare a nerve biopsy specimen for an EM?
    Renal Biopsies

How should we prepare a renal biopsy specimen for an EM study?
    Tumors

How should we prepare a tumor biopsy specimen for an EM study?
    Bone Marrow
How should we prepare a bone marrow specimen for an EM study?
I still have questions…


Research Questions

What is the first step in starting a research project involving the facility?
As a student/researcher, can I learn how to use the TEM? The ultramicrotome? The histology equipment?
I want to send you a sample for EM processing. What do I need to do? What does the facility need to know?
I want to send you a sample for histological processing. What do I need to do? What does the facility need to know?
How much will a research project cost and how much time it will take?
I still have questions…

Atlas Questions

 

General Questions
Why use a transmission electron microscope (TEM)?
Many kinds of questions about high resolution structure can be answered with a TEM including diseased tissue, bacterial and viral structure, cultured cells and manmade materials like support films and particle suspensions. We offer a full line of TEM services but primarily for with biological samples.

Do you have a scanning electron microscope (SEM)?
If you need to use a SEM, contact the University of Toledo's Center for Materials and Sensor Characterization (CMSC) or call 419-530-6080 for more information.

How much tissue is needed for study?
Tissue should be minced into very small cubes no greater than 1mm in the greatest dimension.  If tissues require special orientations (i.e. muscle or cilia) the sample should be no thicker than 1mm in two dimensions while the 3rd dimension should be long enough so that a sample appears as a ‘strip’ of tissue. All tissues must be fixed immediately in glutaraldehyde or glutaraldehyde/paraformaldehyde fixative solution. Fluids for viral characterization do not require fixation.

Why does the sample have to be so small?
The chemicals used to fix tissues for high resolution do not penetrate very fast.

What advantages does the TEM offer over light and/or fluorescence microscopy?
The main advantage that TEM offers is increased resolution over that of microscopy involving visible spectrum light.

How long does it take to process tissue?
Usually 3-5 days but large projects, improper fixation or infiltration, and difficulty sectioning can complicate the process. Our facility is primarily a diagnostic facility and clinical samples take priority over research projects.

What types of TEM procedures are available?
There are many specialized procedures that we offer including negative staining of particles (i.e. viruses, nanoparticles), immunogold labeling and various other techniques. If you are interested in trying some specialized or advanced techniques, please contact us.

Why can’t frozen samples be used for TEM?
Unless special freezing devices are used, ice crystal formation within cells will rupture membranes and cause massive ultrastructural changes.

Why are TEM images in black and white/grey scale?
The electron microscope a beam of monochromatic electrons with an extremely short wavelength; there are no visible wavelength colors to visualize. Color images from electron microscopes have been pseudo-colored using computer software to increase contrast or to help the viewer study the image.

Clinical Application Questions
What clinical electron microscopy services do you offer?
We are a full service diagnostic electron microscopy facility offering:
•    Platelet evaluation for delta granule storage pool deficiency (δ-SPD)
•    Platelet evaluation for structural investigation (i.e. screening for chromosome 11q deletion defects manifesting as giant alpha granules, Gray Platelet Syndrome, Bernard-Soulier Syndrome)
•    Renal pathology
•    Muscle and nerve pathology
•    Tumor ultrastructural morphology
•    Metabolic disease (glycogen storage disease, lysosomal storage processes, drug toxicity)

How should I send my sample?
Detailed instructions for sample submissions including procedures, justification paperwork and mailing addresses are available on our website. Platelet storage pool deficient sample information can be found here. All other TEM submission information can be found here. If you have any other questions on sample submission, please contact us.

Can I send a sample at any time?
No, unfortunately we do not have laboratory coverage on weekends. Our laboratory is staffed Monday through Friday 6:30 AM – 5:00 PM, Eastern time-zone. Blood samples must be received during laboratory hours within 24 hours of venipuncture, therefore any sample requiring express airmail should be sent for next morning delivery. Do not send a blood sample on a Friday.

Should I chill or freeze my sample?
All samples must be sent at room temperature.

How will you bill my sample?
We don’t bill patients or insurance companies. Billing is interinstitutional only. The following form is beneficial for diagnostic test referral justification and can be sent to third party payers: Request for Approval for Platelet Electron Microscopy Form

When should I expect a report?
We attempt to have results within a week but occasionally results may take 2-3 weeks.

If I haven’t received a report, who should I contact?
Please contact our report transcription office at 419-383-3485.

What is delta (dense) granule storage pool deficiency?
Delta granule storage pool deficiency is a disorder that causes common bleeding symptoms due to a reduced number of small storage vesicles contained by the blood platelet.

How many delta granules should platelets have?
Platelets have on average 4-6 dense granules per platelet. Some laboratories have a range of 3-8 dense granules for a normal range but we have established a range of 3.68-6 dense granules per platelet based upon biochemical correlation studies (adenine nucleotide determination) of people without any bleeding history (bleeding checklist questionnaire) or laboratory assessment (CBC and coagulation aggregation study) for our normal range.

What are common symptoms of delta granule storage pool deficiency?
The most common symptoms of delta granule storage pool deficiency include easy bruising, frequent nose bleeds, excessive bleeding with trauma or surgery, excessive bleeding of the gums upon teeth-brushing and for women, heavy menstrual bleeding.

What is the CPT code for a Platelet Storage Pool Deficiency Analysis?
The CPT code is 88348.

How should we prepare and ship a muscle biopsy specimen for an EM study?
A muscle biopsy should be obtained using a Price muscle clamp.  The biopsy site should not be injected with anesthetic and the tissue should not be repositioned once placed in the clamp.  A portion of fresh tissue should be excised from the excess tissue at a cut edge of the biopsy for enzyme histochemistry studies; the fresh tissue should be placed in gauze soaked with saline in a secure sample container.  The clamped tissue should be placed in 3% glutaraldehyde fixative once the tissue for histochemistry has been obtained.  The container should have a tightly fitting cap to prevent fixative spills during transport.  Notify our Histology Laboratory (Telephone number: 419-383-5229) including method of shipment.  Place the specimen container inside a large container packed with crushed ice.  Do not use dry ice to avoid freezing during transport.  Send the specimen by courier following biopsy (our address can be found here).  Please provide the following clinical information: Patient’s age, sex, and site of muscle biopsy, symptoms, signs and duration (including family history), results of EMG study, other relevant clinical laboratory studies, concurrent disease (connective tissue or neoplastic disease).

How should we prepare a nerve biopsy specimen for an EM study?
Place intact (not minced) specimen in 3% glutaraldehyde fixative immediately.  Send the specimen at ambient temperature in a vial containing 3% glutaraldehyde fixative.  Please provide the following clinical information: Patient’s age, sex, and site of biopsy, clinical symptoms, signs and duration (including family history), other relevant clinical laboratory studies, systemic/concurrent disease.

How should we prepare a renal biopsy specimen for an EM study?
Place minced (1mm cubes) specimen in 3% glutaraldehyde fixative immediately.  Send the specimen at ambient temperature in a vial containing 3% glutaraldehyde fixative.  Please provide the following clinical information: Patient’s age, sex, and site of biopsy, clinical symptoms, signs and duration (including family history), other relevant clinical laboratory studies, systemic/concurrent disease.

How should we prepare a tumor biopsy specimen for an EM study?
Place minced (1mm cubes) specimen in 3% glutaraldehyde fixative immediately.  Send the specimen at ambient temperature in a vial containing 3% glutaraldehyde fixative.  Please provide the following clinical information: Patient’s age, sex, and site of biopsy, clinical symptoms, signs and duration (including family history), other relevant clinical laboratory studies, concurrent disease.

How should we prepare a bone marrow specimen for an EM study?
Place bone marrow core specimen in 3% glutaraldehyde fixative immediately. Send the specimen at ambient temperature in a vial containing 3% glutaraldehyde fixative. Include a bone marrow aspirate smear slide. Please provide the following clinical information: Patient's age, sex, and site of biopsy, clinical symptoms, signs and duration (including family history), other relevant clinical laboratory studies, systemic/concurrent disease.



Research Questions
What is the first step in starting a research project involving the facility?
We suggest setting up a short consultation either through email or face to face to discuss your specific research goals and to get you acquainted with our facility. Please feel free to contact us.

As a student/researcher, can I learn how to use the TEM? The ultramicrotome? The histology equipment?
We are more than happy to teach you or a member of your lab how to use our equipment. This may save you time and money (especially if you have a large volume of samples) as we would only charge you for the consumables used. We have specific equipment available that students and researchers can learn and use that is separate from the ones used for clinical samples. No previous experience is need. There is a nominal fee schedule for electron microscope beam time. Please contact us if you are interested in learning to use our equipment.

I want to send you a sample for EM processing. What do I need to do? What does the facility need to know?
The best course of action is to contact us to set up a short consultation to discuss the nuances of sample preparation, embedding, sectioning and staining before you bring your samples to us. We will need to discuss things like sample composition, size, orientation and fixation along with get your contact and billing information.

I want to send you a sample for histological processing. What do I need to do? What does the facility need to know?
The best course of action is to contact us to set up a short consultation to discuss the nuances of sample preparation, embedding, sectioning and staining before you bring your samples to us. We will need to discuss things like sample composition, size, orientation and fixation along with get your contact and billing information.

How much will a research project cost and how much time it will take?
You can discuss research project costs by contacting us anytime. As for time, that may vary depending on our clinical case load, as those cases take priority over research samples. We try and complete every project as timely as possible.

I still have questions….
Please contact us using the information on this page. We will be happy to answer any questions you may have.

Atlas Questions

Last Updated: 3/23/15