We dilute & cytospin fluids depending on WBC counts thusly;

 If a culture is ordered  we usually make 2 slides, 1 for diff & 1 for
gram.

HAEMATOLOGY  MANUAL

AUTHOR:
Technical Coordinator, Core Laboratory
FOLDER:
Body Fluids

APPROVED BY:
Medical Director, Laboratory Services
REVIEW FREQUENCY:
Yearly

RESPONSIBILITY:
Technical Coordinator, Core Laboratory
ORIGINAL

APPROVAL DATE:
03/05/90

REVIEWED/REVISED DATE:
15/02/98

REVISION NUMBER:
2

110.723.005  CYTOSPIN SLIDES FOR BODY FLUIDS

(MSH SITE USE ONLY)

Principle:

The cytospin concentrates cells from a fluid onto a microscope glass
slide using high speed centrifugation.  Only a small amount of fluid is
required.  The cells are distributed in a single layer in an area 6 mm
in diameter.  The slides are air dried for a few minutes, then stained
and examined.

Material Requirements and Preparation:

·         Shandon Cytospin 2

·         2 frosted end glass microscope slides

·         2 reusable cytofunnel sample chambers

·         2 cytoclips

·         plastic transfer pipettes

·         wax pencil

Procedure:

1.       Gently place the rotor bowl into the Cytospin 2 and close the
safety lid.  Set the speed control to 150 (1500 rpm) on HI for 3
minutes.

2.       When spinning has stopped, remove the sample chamber assembly.
Carefully remove the slide.

3.       Leave one slide as concentrated smear.  Before the second one
dries, take a clean slide and spread out concentrated cells.  This
allows for undistorted morphology in very concentrated Label 2 glass
microscope slides with the computer labels of the corresponding
patient.

4.       Place slide behind the cytofunnel sample chamber with the
attached filter paper.  Make certain the bottom of the slide matches up
with the bottom of the sample chamber.  Draw a circle on the slide
around the sample area using a wax pencil.

5.       Place the slide and cytofunnel sample chamber into the
cytoclip and close by bringing the spring clip up and holding it into
place using the retaining hooks.

6.       Remove rotor bowl from Cytospin 2 and remove the lid.  Place
sample chamber into the rotor bowl.  Pipet samples into the sample
chamber, cap and replace the lid while the rotor bowl is outside the
Cytospin 2.

7.       Guideline for fluid dilutions for Cytospin preparation slides
to be done on all body fluids.

WBC Count             Ratio     Drops of Fluid    Drops of CD3500
Diluent

X10 9/L

0    - 0.1                                          20
  ---

0.1 - 0.3                  1:2                    10
10

0.3 - 0.7                  1:4                      6
18

0.7 - 1.5                  1:10                    2
18

1.5 - 3.0                  1:20                    1
19

3.0 - 5.0                  1:30                    1
20

8.       Place 5 drops of diluted fluid into sample chambers.

9.       Centrifuge specimens.

10.   Air dry slides, then stain in the routine manner.

11.   Save all fluids in the refrigerator for 7 days.

12.   Disinfect the useable cytofunnel sample chambers in 1%
hypochlorite for 24 hrs.

NOTE:  For troubleshooting aberrant results refer to the Shandon
Cytospin 2 manual under Qualitative Determinants, page 14.





References:

1.       Shandon Cytospin 2 manual