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Principles of specimen collection in pathology. Jumphol Mitchai MD Department of pathology KKU. Pathology services. Surgical pathology Cytopathology Exfoliative cytology Body fluids Fine needle aspiration (FNA, FNAC, FNAB ) Cytogenetics Autopsy . Surgical pathology.

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principles of specimen collection in pathology

Principles of specimen collection in pathology

Jumphol Mitchai MD

Department of pathology KKU

pathology services
Pathology services
  • Surgical pathology
  • Cytopathology
    • Exfoliative cytology
    • Body fluids
    • Fine needle aspiration (FNA, FNAC, FNAB)
  • Cytogenetics
  • Autopsy
surgical pathology
Surgical pathology
  • Examined all tissues and foreign objects removed from living patients
  • Specimens not to be submitted must be written as hospital policy
  • Practice of pathologists
  • Pathological diagnosis : “ Gold standard in medicine ”
specimen collection in surgical pathology
Specimen collection in surgical pathology
  • Incisional biopsy : for diagnostic purpose
  • Excisional biopsy : for diagnostic and therapeutic purposes
  • Tissue collection by instruments : needle biopsy, core biopsy, punch biopsy, curettage, shave biopsy, Bard biopsy, endoscopic biopsy
  • Necropsy = biopsy from dead body
  • Organs from operation rooms
general rules for the biopsy
General rules for the biopsy
  • Larger the lesion - more numerous biopsies
  • Ulcerated tumor – recommend peripheral biopsy
  • Avoid necrosis and hemorrhage areas
  • All fragments must be sent to the pathology department
  • Not crushing, squeezing, cautery
  • Immediately fix in fixatives
  • Orientating specimen e.g. deep margin, superior and inferior margins by using sutured threads (long, short)
  • Transparency
  • Lid
  • Leak-proof
  • Wide mouth, flat bottom
  • Size corresponding to specimen volume
  • Label
fixation of the tissue
Fixation of the tissue
  • Hard tissues take more time to thoroughly fixed than soft tissue.
  • Penetration rate, around 1 mm./hr. in general (variable, depends on consistency of the tissue).
  • 10 times volume of fixative over the specimen volume is needed.

Neutral buffered formalin - fixation time 12-24 hours.

  • Formalin (40% aqueous solution of formaldehyde) - 100ml
  • Sodium dihydrogen orthophosphate (monohydrate) - 4g
  • Disodium hydrogen orthophosphate (anhydrous) - 6.5g
  • Distilled water - 900ml

This fixative is suitable for most histological purposes. It is to be preferred to formol-saline (a single 10% solution of formalin in 9% aqueous NaCl) as formalin pigment is avoided. Specimens may be stored in this fluid. The solution is isotonic.


1 การติดสลากบนภาชนะใส่ข้อมูลผู้ป่วยให้ครบถ้วน ชื่อสกุล HN เพศ อายุ แพทย์เจ้าของไข้ หอผู้ป่วย


2 การเขียนใบนำส่ง


specimen(organ), method of specimen collection, characteristics, clinical informations)

filling the request forms 1
Filling the request forms (1)

1. identified of the patient

2.identified specimen ( gross appearance, cyst, vascular, inking for margin) type of surgery ( biopsy, wide excision) surgical finding

3. adequate clinical history(necessary)

prior diagnosis, immune status (HIV), prior treatment : radiation, chemotherapy bone lesion : X-ray finding

filling the request forms 2
Filling the request forms (2)

bone lesion : X-ray finding hematolgic disease: CBC,bone marrow

female reproductive lesion : LMP, hormone

hepatitis : LFT4. Clinicaldiagnosis or differential diagnosis

immunohistochemistry and her2 study
Immunohistochemistry and HER2 study
  • Use formalin-fixed paraffin embedded tissue (paraffin block)
  • Must have request forms
frozen section
Frozen section
  • Intraoperative consultation
  • Fresh tissue was immediately freezed in liquid nitrogen and make an ultrathin section in cryostat
  • Frozen section is not a permanent section
  • Cryostat = microtome in the freezing cabinet.
indications for frozen section
Indications for frozen section
  • Benign or malignant lesions
  • Adequacy of surgical margins
  • Metastatic lymph node : sentinel nodes in breast carcinoma
  • Presence or absence of ganglion cells in large intestinal wall of Hirschsprung disease
diagnosis of frozen section
Diagnosis of frozen section
  • Benign VS malignant : not specific diagnosis
  • In case of doubt – wait for permanent section
specimens for frozen section
Specimens for frozen section
  • Fresh tissue
    • < 1cm. in size is recommended.
  • Put in plastic bag, tightly closed with rubber bands
  • Label on the plastic bag
  • Place the plastic bag in ice
  • Send it to pathology lab immediately
inappropriate specimens for frozen section
Inappropriate specimens for frozen section
  • Large tumor or specimens
  • Highly infectious specimens : TB, HIV
  • Follicular lesion of thyroid (can not diffentiate between benign and malignant)
  • The study of individual cells or small clusters of cells for diagnosis and screening of diseases, including cancers
  • practice of screeners, cytotechnicians, cytopathologists
specimen collection in cytopathology
Specimen collection in cytopathology
  • Specimens
    • Smear slide
      • Cervix (Pap smear)
      • Fine needle aspiration (FNA)
      • Bronchial brush
    • Fluid
      • Sputum
      • Bronchial wash
      • Body fluids, urine, CSF
วิธีการเตรียมสิ่งส่งตรวจทาง cytopathology

1 สิ่งส่งตรวจที่ป้ายบนสไลด์ให้แช่ใน95%Alc. ทันที

2 Fluid specimen ให้นำส่งห้องlabทันที หรือเก็บในตู้เย็นได้นาน12-24ชม.

3 เขียนใบนำส่งที่มีรายละเอียดของผู้ป่วย และประวัติการเจ็บป่วย

4 ติดสลากระบุข้อมูลผู้ป่วยบนภาชนะหรือสไลด์

5 ส่งห้องปฏิบัติการทันที

  • Study structure of chromosome from cell culture
  • FISH in chromosome of leukemia
  • Specimens
    • Whole blood
    • Bone marrow
    • Amniotic fluid
specimen collection for cytogenetics
Specimen collection for cytogenetics
  • Sterile techniques
  • 2-5 ml. of blood or bone marrow in heparinized tube or sterile heparinized syringe
  • 20 ml. of amniotic fluid in sterile disposable syringe
  • Send to lab immidiately (with request form) or keep in refrigerator not more than 12 hours
service hours for cytogenetics
Service hours for cytogenetics
  • For blood specimen : Mon, Tue, Fri 9.00-16.00
  • For Bone marrow : Mon, Thur 9.00-16.00
  • For amniotic fluid : Tue 9.00-15.00
  • Blood and bone marrow for FISH : Mon-Fri 9.00-16.00

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