Michener TITLE: SECTION:4

Simulated Health Cytocentrifugation

Network CLCY-4-03

DIAGNOSTIC LABORATORY STATUS: VERSION: 3 Page 1 of 3

CYTOLOGY MANUAL APPROVED

1.0 Purpose:
 In Diagnostic Cytology, the cytocentrifuge is used to separate cells from a small amount
of fluid.
 This procedure may be used for all body cavity fluids, urines, bronchial wash or any fluid
specimen where the volume is too small to produce a pellet or sediment, after routine
centrifugation.
 It is the method of choice for the preparation of CSF samples. Urine samples may also be
processed with this method for maximum cell retention.

2.0 Scope:
 Cytospin (or cytocentrifugation) utilizes a technique where the cells are concentrated
onto a 6 mm diameter section of a slide for rapid screening with minimal cell loss.
 Used for hypocellular samples or samples with volumes less than 5 mL

3.0 Definitions:
 Centrifugation: using centrifugal force to separate substances of different density or
particle size, when suspended in a fluid, by spinning them about an axis.

4.0 Related Policies/Procedures:

 Laboratory Safety CLCY-2-01
 Centrifugation CLCY-4-02
 Cytocentrifugation Quality Control Record CLCY-9-09

5.0 Equipment: 5.1 Supplies 5.2 Reagents

 Laboratory Centrifuge  Disposable pipettes  Cytospray or

 Cytocentrifuge
 Vortex
 Biological Safety Cabinet
 PPE – Lab coat, gloves
 Disposable Centrifuge Tubes  95% ethyl alcohol
(15ml)
 Cytofunnels with lids
 Cytoclips
 Coplin Jar(s)
 Cytospin slides
 Forceps
 Pen / Pencil / Grease pencil
 Paper Towel

6.0
PROCEDURE WORK INSTRUCTIONS RATIONALE

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge May 03, 2010
Revised March 08, 2017
 Michener TITLE: SECTION:4
Simulated Health Cytocentrifugation
Network CLCY-4-03

DIAGNOSTIC LABORATORY STATUS: VERSION: 3 Page 2 of 3

CYTOLOGY MANUAL APPROVED

STEPS
6.1 Getting 1. Treat all samples as if they were infectious. Infection
started 2. Wear appropriate PPEs – lab coats and gloves. control.
3. Turn on Biological Safety Cabinet (BSC) and allow air
to stabilize for 5 minutes.
4. Take samples from refrigerator/designated area and
place in the BSC once they are accessioned.
5. Allow samples to come to room temperature.
Specimens are
6. Label appropriate sized test tube with accession centrifuged
number using grease pencil prior to being
6.2 Centrifugation 7. Centrifuge the specimen (see CLCY-4-02) cytocentrifuged
8. Pour off supernatant, leaving a few drops of
supernatant, and re-suspend the cell button (using a
6.3 Cyto-
vortex mixer or disposable pipette)
centrifugation 9. Gather required cytospin equipment in BSC:
cytofunnels with lids, cytoclips, glass slides, rotor
bucket and lid.
10. Assemble the slides and cytofunnels into the cytoclips.
11. Place a few drops of the centrifuged specimen into the
cytofunnel. (On average 2 drops per cytofunnel) It is essential to
12. Secure small plastic lids on cytofunnel. remove
13. Place the cytoclip apparatus into the cytospin specimens
rotor/bucket. carefully but
quickly from the
14. Be sure to balance the cytospin with equal numbers of cytospin to
samples placed opposite each other in the rotor/bucket. prevent air
15. Secure the lid on the rotor/bucket. drying
16. Spin specimen for 4 minutes at 600 rpm.
17. When cytospin comes to a complete stop, remove
rotor/bucket and place in BSC (Biological Safety Tilting the
Cabinet) assembly
18. Remove slides by carefully by undoing the cytoclips toward the
and pulling the cytofunnel gently away from the slide funnel side
as not to disrupt the cell preparation. prevents any
residual
19. Place slides directly into 95% ethanol fixative for 30 supernatant
minutes. from washing
20. Place disposable cytofunnels and materials into away the cells
biohazardous disposal bag. on the slide
21. Place reusable equipment (cytoclips) into disinfectant

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge May 03, 2010
Revised March 08, 2017
 Michener TITLE: SECTION:4
Simulated Health Cytocentrifugation
Network CLCY-4-03

DIAGNOSTIC LABORATORY STATUS: VERSION: 3 Page 3 of 3

CYTOLOGY MANUAL APPROVED

for 15 to 20 minutes.
22. Wash equipment and dry thoroughly.
23. Clean and disinfect centrifuge and cytocentrifuge after
each use
24. Remove contaminated gloves and wash hands.
25. Follow staining procedure when fixation time is
complete

7.0 Documentation:
 File requisitions in appropriate lab binder.
 Enter data into Laboratory Daily Log Book
 Complete all Quality Control forms.

8.0 References:
 Bibbo M., Wilbur D., Comprehensive Cytopathology, 3rd ed., Elsevier Inc, 2008.
 https://www.fishersci.ca/shop/products/cytospin-4-cytocentrifuge-cytospin-4-2/29618304
 The Michener Safety Manual (current version).

9.0 Developed in Consultation With:

Catherine Brown Jane Mattson
Eileen McDonald Mary Davis

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge May 03, 2010
Revised March 08, 2017