Pathology Submissions
Special Stains
NecropsyMaximum number of animals (similar age and clinical problem) to be included in 1 accession:
Cattle: 1 adult or 2 calves
Cats: 1 adult or 3 kittens
Dogs: 1 adult or 3 puppies
Horses: 1 adult or 2 foals
Sheep: 1 adult or 3 lambs
Swine: 1 adult or 3 pigs
Abortion/Stillbirth: 3 fetuses w/placenta
Shipping containers for mail or courier services must be leak-proof and airtight-All species

-Carcasses must be held at refrigeration temperature until delivery to ADDL.

-DO NOT freeze unless interval between death and delivery > 3 days.

-Carcasses smaller than 150lbs can be delivered after hours to the walk-in refrigerator on the south dock (be sure to place red hold tag with owner's name, date and contact information on animal before placing in white cooler).

-After hours, if delivering an animal over 150lbs and will not fit in the white cooler please contact ADDL at 765-494-7440 to be put in contact with the pathologist on duty.

-Gross necropsy results will be reported in 1-5 working days; final reports are completed within 5-14 working days.

-ADDL does not euthanize companion animals, including horses.
Cosmetic necropsy1 animal-Prior arrangements with ADDL are required. However, cosmetic necropsy is discouraged because it prevents thorough postmortem examination and may hinder diagnosis.

-Animal must be in ADDL before 1 pm and will usually be returned to owner the same day. Additional charge applies.
Legal/Insurance Necropsy1 animalMust be designated as such prior to necropsy. Additional charge applies.
Field ConsultationCall to schedule. Field consultations are made only at the request of a veterinarian.
Practitioner-performed necropsy-Brain: whole or half

-Spinal cord: entire, with opened dura

-Eyes: whole

-Endocrine glands: small, whole; larger glands, bisected

-Heart, lung, liver, kidney, spleen: 5 mm thick slices (whole heart if cardiac defect is suspected)

-GI tract: 3-5 cm segments
10% formalin-Brain, heart, lung, liver, kidney and spleen plus any other tissues with gross lesions should be submitted from all necropsy examinations. Other tissues should be included according to history or clinical signs.

-Formalin-fixed specimens accompanied by fresh (unfixed) specimens for ancillary testing (microbiology, toxicology, virology etc) are processed as long-form accessions. Histologic findings are reported in 3-5 working days with final reports in 7-15 days.
Amputations-Small specimens, e.g. digits, may be submitted whole

-Larger specimens, e.g. limbs, should be refrigerated and delivered to ADDL within 24 hours
10% formalin -Refrigeration for large specimensBony specimens will require 1-10 day delay for decalcification. Additional charges apply.

-Limb amputations can be submitted as a "Biopsy Plus" for dissection and gross examination by a pathologist before histopathology. These will be charged as a necropsy.
HistopathologyVarious tissue specimensLeak-proof wide-mouth jar or Whirl Pak-Tissues should be fixed in 10% buffered formalin with 10:1 formalin to tissue ratio.

-Excess formalin can be decanted after 1-2 days fixation.
Endoscopic, pinch, core (Tru-cut), or punch biopsiesEntire biopsy specimen: if <3mm in any dimension, submit in histologic cassette to avoid specimen damage or loss10% formalinMultiple specimens should be submitted in individually labeled containers to indicate tissue site. Histologic findings from Neoplasm Submission Form accessions are reported in 1-3 working days. Histochemistry or immunohistochemistry requires another 1-5 working days.
Excisional biopsiesIf >2cm in diameter, slice accordingly to allow fixation10% formalinUse paint or suture tags to orient specimen or identify margins, if pertinent. Histologic findings from Neoplasm Submission Form accessions are reported in 1-3 working days.
Histopathology Submission Information
  • Please complete an appropriate Submission Form, available on the Forms page, and submit with sample.
  • Tissue samples should be no more than 0.5 cm thick and placed in a wide mouth jar containing 10% buffered formalin fixative.
  • For fixation, the formalin amount should be at least 10X the volume of the sample.
  • Alternatively, tissues may be fixed for 24-48 hours and then transferred to a Whirl Pak bag with enough formalin to prevent drying.
  • Tissues should arrive at ADDL already fixed in formalin.
  • If formalin is not used as the fixative, please indicate which fixative was used.
  • Before shipping, contact the postal service or courier service to be sure to comply with their regulations.
  • Histopathology mailers are available from ADDL. See the Supplies page.
Painting the Surgical Margins of a Tumor Biopsy
A common problem faced by pathologists is interpreting surgical margins of tumor biopsies. Although it is often easy to distinguish surgical margins from those produced during the trimming process in the histology laboratory, it is sometimes difficult and frustrating for both pathologists and clinicians. Diagnosticians know that the three most common questions posed by the clinician regarding a neoplastic process are "What kind of tumor is it?," "Is it benign or malignant," and "Are the margins free of neoplastic cells?"
In human medicine, surgical margins of a biopsy are commonly painted with a dye that adheres to tissue and is visible under the microscope. The procedure is simple and does not interfere with histologic evaluation. Its advantage is that it clearly distinguishes surgical margins from trimming margins. This is essential to assess complete/incomplete excision of a tumor.
Biopsy margin painting can be done on unfixed or fixed tissues; however, painting unfixed tissues is easier and preferred. There are several commercially available products for this purpose (call the ADDL for additional information, if needed). The use of different colors for different aspects of mass orientation is superior to using sutures of different colors. Painting biopsy margins is also inexpensive; a 20 ml bottle of dye will last several years.
Please check the Winter 2004 Newsletter for additional information.
How to Paint the Surgical Margins of a Tumor Biopsy
  1. Blot the biopsy margins of the mass.
  2. Select the dye color.
  3. With a wooden applicator stick or cotton swab, "paint" the biopsy margin.
Do not pour dye on the surface; apply as if painting.
  1. Let the dye dry for 5-10 minutes, and immerse the sample in regular fixative solution. Some of the dye will dissolve with the fixative; this will not affect the sample. If the sample is thicker than 4-6 mm, section it to improve penetration of fixative as you usually do.
Alcian Blue pH 0.4
Alcian Blue pH 1.0
Alcian Blue pH 2.5
Alican Blue/PAS
Brown & Brenn
Brown & Hopps
Congo Red
Gomori One-step Trichrome
H & E
Jones PASM
Luxol Fast Blue
Melanin Bleach
Modified ZN (lipofuschsin)
Movat Pentachrome
Oil Red O Fat Stain
PAS (Periodic Acid-Schiff)
PAS/ Diastase
Picrosirius red
Prussian Blue
Rubeanic Acid
Toluidine Blue
Trichrome (Masson)
Van Gieson
Verhoeff's (VVG)
Von Kossa
Warthin Starry
Ziehl-Neelson - Acid Fast