Handling renal biopsies
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Renal biopsy (also kidney biopsy) is a medical procedure in which a small piece of kidney is removed from the body for examination, usually under a microscope. Microscopic examination of the tissue can provide information needed to diagnose, monitor or treat problems of the kidney.
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Handling renal biopsies
- 2. Indications of renal biopsy •Unexplained acute or rapidly progressive renal failure •Nephrotic syndrome and significant non-nephrotic proteinuria •Persistent glomerular hematuria •Systemic diseases with renal involvement •Renal allograft dysfunction.
- 3. Required Information Label the containers with the patient’s name and a 2nd patient identifier such as a date of birth or a medical record number. Detailed clinical history should be provided by the clinician.
- 4. RENAL BIOPSIES LIGHT MICROSCOPY IMMUNOFLUORESCENCE ELECTRON MICROSCOPY IMMUNOHISTOCHEMISTRY
- 5. Adequacy The minimum diagnostic sample size varies with the specific diagnosis; for instance only one glomerulus is enough for making a diagnosis of membranous glomerulonephritis while 25 glomeruli may be required to make an accurate diagnosis of focal lesion like FSGS. For most of the light microscopic assessment, 8 - 10 glomeruli are considered adequate.
- 6. Adequacy The adequacy of the renal tissue core should be assessed on - site using a dissecting microscope. The tissue sample should be transferred using a wooden spatula to a glass slide with few drops of saline and examined. Under a dissecting microscope, glomeruli appear as reddish, circular structures while medulla is identified by red streaks running almost parallel.
- 8. If possible, 2 - 3 cores should be taken: one for light microscopy (LM), another for immunofluorescence(IF) and one for electron microscopy(EM), if required. In cases where taking extra passes is not possible, a cutting protocol may be followed: both the ends of the core are taken for EM, one-third of the core, including some glomeruli, is placed in transport medium for IF and the rest is kept for LM.
- 11. Precautions while dividing renal tissue cores Forceps should not be used, as they may lead to crush artefacts. Thin wooden stick, for example, a spatula or toothpick, is ideal. Avoid touching the tissue with a fixative-contaminated scalpel or blade; this may contaminate the tissue meant for IF.
- 12. LIGHT MICROSCOPY The tissue sections should be no greater than 2-3 μm in thickness Good histology also demands good fixation. If fixation is delayed and imperfect, it cannot be improved later. The most commonly used fixative for LM is buffered, 10% aqueous formaldehyde solution (formalin). Some laboratories prefer alcoholic Bouin’s or Zenker’s fixatives for better morphological details. However, these interfere with recovery of material for EM, IHC or molecular studies.
- 14. IMMUNOFLUORESCENCE Immunofluorescence is best performed on unfixed, frozen sections. Tissue can be transported to the laboratory fresh on saline-soaked gauze or in Michel's fixative. Serial sections are cut at 2-4 μm in a cryostat. IgG, IgM, and IgA, C3, C1q, C4, fibrin, and kappa and lambda light chains.
- 15. Electron Microscopy The tissue for EM may be fixed in 2-3% glutaraldehyde or 1-4% paraformaldehyde. Adequate fixation can also be obtained when tissue is fixed in buffered formalin. EM cannot be performed on tissues exposed to mercury-based fixatives (e.g., Zenker's). Tissue can be reprocessed from the paraffin or the frozen block if no glomeruli are available in the EM sample.