> **Generic template - SUMMARY OF PROCEDURE FOR [KIDNEY
> BIOPSIES]{.underline}**
>
> The **kidney biopsy** (needle or open wedge) requires special
> handling. Materials and reagents required include (1) a specimen
> container with buffered formalin, (2) a specimen container with 3%
> glutaraldehyde in 0.1M sodium cacodylate buffer, (3) a specimen
> container with immunopathology fixative (Michel\'s fixative; Zeus
> medium), (4) a bottle of phosphate buffered saline (PBS), (5) a
> specimen container with a 2 X 2 gauze, (6) a pair of forceps, (7) a
> single edge razor blade, (8) a sheet of wax, and (9) a VA Surgical
> Pathology Request Form (SF 515).
>
> The biopsy should first be placed on the 2 X 2 gauze previously
> moistened with PBS to prevent desiccation, placed in a clean, empty
> specimen container, and transported as rapidly as possible to the
> Surgical Pathology Laboratory.
>
> Once in the laboratory, the biopsy should immediately be divided and
> fixed as follows: Place the cores on the sheet of wax in several
> droplets of PBS; be careful to keep PBS on the biopsies at all times.
> Place under a dissecting microscope and divide according to the scheme
> illlustrated.
>
> **a. Tissue for Immunofluorescence/Immunoperoxidase Studies**
>
> A portion of the central region of the core(s), from the cortex and
> ideally containing one or more glomeruli, should be excised for
> immunostaining. The specimen should be placed in the immunopathology
> (Michel\'s) fixative and kept at room temperature. Care must be
> exercised at this point to avoid contamination of the specimen with
> formalin or glutaraldehyde, as this precludes successful
> immunofluorescence or immunoperoxidase analysis.
>
> **b. Tissue for Electron Microscopy**
>
> One to four very small (1 mm) sections of renal cortex, ideally also
> containing glomeruli, should be cut from the ends of the biopsy core
> and placed in the bottle of glutaraldehyde for ultrastructural
> studies. Buffered formalin is an acceptable although not ideal
> alternative to glutaraldehyde for fixation and shipping.
>
> **c. Tissue for Routine Surgical Pathology**
>
> The remaining tissue should be placed in the specimen container with
> formalin for conventional paraffin embedding and histological
> examination by light microscopy.
>
> If a limited amount of tissue is available, priority should be given
> to the portion allocated for routine surgical pathology.
>
> Label all containers with the patient\'s name and social security
> number. On the accompanying SF515, include information regarding the
> type of biopsy (medical, transplant, or transplant donor; needle or
> wedge), pertinent clinical history (including current medications),
> and who should be contacted with results (including phone and/or Pager
> ID numbers).
>
> Make sure all caps on vials are secure and seal with parafilm.
> Transport or mail all specimens to:
>
> **Surgical Pathology Laboratory, Room XXXX**
>
> **Pathology and Laboratory Medicine Service, 113**
>
> **Local VA Medical Center**

##  000 Street

> **City, State Zip**
>
> Be sure to label package on outside as "URGENT: PATIENT SPECIMENS".
> Specimens obtained after hours or on weekends should be held in the
> appropriate fixatives and mailed on the next work day.
>
> Address questions concerning submission of renal biopsies or other
> materials to: Local Names/contacts/addresses/phone numbers.
>
> **REAGENTS FOR KIDNEY BIOPSIES**

**1. Electron Microscopy**

**Buffer**

**[0.2 M Sodium Cacodylate Buffer]{.underline}**

42.8 gm. Na cacodylate

fill to 1000 ml. with distilled H2O

Adjust pH to 7.4 using 37% HCl

Osmolality 262 mOsm.

Store in refrigerator for one year.

**Fixative**

Glutaraldehyde comes in 25% sealed glass ampoules. Keep refrigerated.

Amount of stock glut. to use =

[% desired x volume desired]{.underline}

\% stock

Half of the final volume should be 0.2 M Na Cac. buffer so final
concentration is 0.1 M.

For 100 ml of 3% made from 25% stock:

[3 x 100 m]{.underline}l- = 12 ml.

25

to 12 ml. 25% glutaraldehyde

add 50 ml. 0.2 M Na cacodylate buffer

and 38 ml. distilled H2O

pH should be 7.2

osmolality 600-650 mOsm.

> Store in refrigerator for one month.
>
> Label with name of solution and concentration, pH, osmolality,
> preparation date, expiration date and initials of person preparing
> solution.
>
> **2. Immunofluorescence**
>
> Michel's Fixative\--available commercially from Wampole/Carter
> Wallace, P.O. Box 1001, Cranbury, NJ 00512; 800-257-9525; Vendor Stock
> \# 0102; 10 ml/bottle, 12 bottles/case; purchase by the case ;
> Michel's Washing Solution, Vendor Stock# 0103.
>
> **3. Light Microscopy (Conventional Histology)**
>
> 10% Formalin is available commercially from Fisher Scientific or
> others.