Principles of specimen collection in pathology
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Principles of specimen collection in pathology

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Pathology services. Surgical pathologyCytopathology Exfoliative cytologyBody fluidsFine needle aspiration (FNA, FNAC, FNAB)CytogeneticsAutopsy . Surgical pathology. Examined all tissues and foreign objects removed from living patientsSpecimens not to be submitted must be written as hospital
Principles of specimen collection in pathology

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1. Principles of specimen collection in pathology Jumphol Mitchai MD Department of pathology KKU

2. Pathology services Surgical pathology Cytopathology Exfoliative cytology Body fluids Fine needle aspiration (FNA, FNAC, FNAB) Cytogenetics Autopsy

3. 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 ?

4. 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

5. 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)

6. Containers Transparency Lid Leak-proof Wide mouth, flat bottom Size corresponding to specimen volume Label

7. 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.

8. Fixatives 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.

9. ???????????????????? 1 ??????????????????????????????????????????? ???????? HN ??? ???? ??????????????? ????????? ?????????????????????? 2 ??????????????? ????????????????????????????????????? specimen(organ), method of specimen collection, characteristics, clinical informations)

11. 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

12. Filling the request forms (2) bone lesion : X-ray finding hematolgic disease : CBC, bone marrow female reproductive lesion : LMP, hormone hepatitis : LFT 4. Clinical diagnosis or differential diagnosis

15. Immunohistochemistry and HER2 study Use formalin-fixed paraffin embedded tissue (paraffin block) Must have request forms

16. 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

17. Cryostat Cryostat = microtome in the freezing cabinet.

18. Frozen sectioning

19. 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

20. Diagnosis of frozen section Benign VS malignant : not specific diagnosis In case of doubt ? wait for permanent section

21. 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

22. 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)

23. Cytopathology The study of individual cells or small clusters of cells for diagnosis and screening of diseases, including cancers practice of screeners, cytotechnicians, cytopathologists

24. Specimen collection in cytopathology Specimens Smear slide Cervix (Pap smear) Fine needle aspiration (FNA) Bronchial brush Fluid Sputum Bronchial wash Body fluids, urine, CSF

30. ??????????????????????????? cytopathology 1 ?????????????????????????????????95%Alc. ????? 2 Fluid specimen ????????????lab????? ???????????????????????12-24??. 3 ????????????????????????????????????? ????????????????????? 4 ???????????????????????????????????????? 5 ??????????????????????

33. Cytogenetics Study structure of chromosome from cell culture FISH in chromosome of leukemia Specimens Whole blood Bone marrow Amniotic fluid

34. 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

35. 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

36. ????????????????????????? ???????? ???HN ????????? ???????????????????????????????????? HN??????????????????????????? ????????????????????, ??????????????? ???????????????????????????????????????????? ?????????????????????????????????? ??????????????????????

37. Thank you


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