The Ohio State University Health System
Department of Pathology – Renal/EM Lab
Weekend coverage for C4d immunostaining
Written By:
Gyongyi Nadasdy M.D.
Date Adopted:
9/10/03
Supersedes Document #:
Review Date:
Revision Date:
Signature:
PRINCIPLE:
Immunofluorescence (IF) for C4d is a sensitive test to diagnose humoral allograft
rejection and became an important diagnostic test in transplant pathology. Therefore, C4d
IF should be provided on a continuos (“rush”) basis, particularly following the
introduction of a protocol to transplant high-risk presensitized (cross-match positive)
patients.
SPECIMEN COLLECTION: The nephrologist or transplant surgeon contacts the surgical pathology
resident on call, that a rush transplant biopsy will be delivered to Surgical Pathology for
“stat” light microscopy and “stat” C4d test processing. The resident on call will arrange
the specimen pick-up according to the Gross Room protocol. After the specimen has
arrived the resident contacts the Renal Pathologist (Drs. Tibor Nadasdy Pager: 303-8625
or Daniela Proca Pager: 730-6212) on-call. The biopsy needs to be triaged according to
protocol using the “Renal biopsy triage sheet”. After the specimen is triaged the resident
contacts the histotechnologist on-call [(614) 303-8821) and the IF technician on-call
[Pager: (614) 637-7151] for “stat” processing. The tissue in Zeus Scientific transport
medium accompanied with a copy of the requisition slip should be placed in the IF-Renal
bucket by the accessioner’s desk in the gross room.
SPECIMEN PICK-UP: The IF technician on-call comes to the gross room (Doan Hall E415) to pick up
the specimen with the requisition slip from the IF bucket.
TEST PROCEDURE: This contains only the list of the steps for preparation of the C4d stain. For more
detail see individual procedures including, “Handling tissue specimens Received in Zeus
Scientific Tissue Fixative”, “Freezing Tissue Specimens for Immunofluorescence
Staining”, “C4d Immunofluorescence Staining For Frozen tissue”
1.
2.
3.
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Tissue received in Zeus Scientific Tissue fixative needs to be washed from the
fixative (Wash solution is located at the IF working area left drawer).
Snap freeze OCT (counter top or in the left drawer of the IF working area) embedded
tissue in Liquid Nitrogen (30 liter dewar is located under the counter opposite to the
IF work area).
Cut 2-3 m sections and pick them up to positively charged slides. 3 sections on two
slides. One should be fixed in 95% alcohol and stained with HE, the other should be
air dried for ½ hour and stained for C4d.
Renal/EM Lab
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The Ohio State University Health System
Department of Pathology – Renal/EM Lab
4.
Do C4d stain. All antibodies, solutions with the exception of PBS are located in the
left refrigerator door in a box labeled “C4d kit”. PBS for IF is on the top shelve of
the IF work are in one gallon plastic container (additional containers are located in
the back of the Immuno lab where the microscope is).
SLIDE DELIVERY: Slides in a covered folder with the requisition slip attached are placed on the right
bottom shelve of the left refrigerator by the ultra low freezer in the Immunopathology
laboratory (N322 Doan Hall). The Renal Pathologist on-call is paged simultaneously.
PARTICIPATING TECHNOLOGISTS: (as of 9/3/2003)
Anna Cherry
Mary Marin
Deborah Rhodes
Nancy White
ON-CALL SCHEDULE: The schedule is developed at least 3 mo. in advance and posted in the EM
Laboratory. Each participant is delivered a copy of the schedule. After the schedule has
been distributed in case the on-call person assigned to a certain date can not fulfill their
obligation it is their responsibility to find coverage. If a schedule change is made it has to
be changed on the EM Laboratory copy.
COVERED HOURS: Saturday, Sunday and holidays the technicians are standby on-call: 8:00 AM2:00PM. That means if a specimen is received before or at 2:00PM and the C4d test is
requested the technician has to come in and complete the test the same day. If the
specimen arrives after 2:00 PM, the IF technician on call has to be notified, but she does
not have to come in the same day, the staining will be done the next morning.
PAGER AND KEY PICK-UP: The on-call pager and keys can be picked-up on Friday or the day before
the holiday from the EM Laboratory (Starling-Loving Hall M018) till 5:00 PM and shall
be returned the following workday or Monday. Special arrangements are made for Nancy
White.
TIME REPORTING: A time stamped overtime sheet is filled out and turned in to Gyongyi Nadasdy (in
her absence Edward Calomeni) the following morning after the holiday or Monday.
REIMBURSEMNT: For technicians on hourly pay the reimbursement follows the standard on-call pay
schedule. For technicians on salary special fee is calculated.
IMPORTANT PHONE NUMBERS:
C4d on-call pager: 637-7151
Histotechnologist on-call pager: 303-8821
EM Laboratory: 293-9258
Renal Pathologists:
Tibor Nadasdy MD. : Pager 303-8625;
Daniela Proca MD. : Pager 730-6212
Laboratory technicians:
Anna Cherry: (937) 642-7185
Mary Marin: 451-8911
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Renal/EM Lab
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The Ohio State University Health System
Department of Pathology – Renal/EM Lab
Deborah Rhodes: 574-6474
Nancy White: 891-4484
QUALITY CONTROL: See “Quality Assurance” for the Immunopathology Laboratory.
REFERENCES: Procedure developed in-house
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