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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

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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.

Principles of specimen collection in pathology


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.

Frozen sectioning

Frozen sectioning

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

Principles of specimen collection in pathology


ชื่อสกุล และHN ไม่ชัดเจน



ไม่ระบุหน่วยงานนำส่ง, แพทย์เจ้าของไข้




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