Shandon Cytospin 4 Standard Workflow Diagram
SPECIMEN EXAMINATION
Page 89
- Specimen origin – precise anatomical size
- Volume of specimen
- Physical appearance of specimen – colour, viscosity, whether it is homogenous or contains tissue fragments or blood
DETERMINE THE CELL COUNT
Page 90
The concentration chosen should allow enough space for cells to form a monolayer with minimum overlap, but not leave too much space between cells.
Average cells (diameter 10-12 microns) produce excellent Cytospin preparations at cell densities of 1×106 cells per ml. Larger cells will require lower concentrations; smaller cells, cell organelles or bacteria will require higher concentrations.
CONCENTRATE OR DILUTE THE SPECIMEN AS REQUIRED
Page 94 (to concentrate);
Page 95 (to dilute)
LOAD SHANDON CYTOSPIN SAMPLE CHAMBERS
Page 40-44 and page 95
Place the EZ Cytofunnels into the sealed head. Make sure they are distributed evenly so that the Shandon Cytospin 4 is not out of balance.
Load the EZ Cytofunnels after they have been inserted into the sealed head.
Do not place more than 0.5ml of a sample in an EZ Cytofunnel sample chamber with white filter card (0.4ml for an EZ Cytofunnel sample chamber with brown filter card or 6ml maximum in an EZ Megafunnel). Make sure that the sample is deposited directly into the bottom of the sample chamber – do not allow the sample to drip down the sides of the chamber.
BE AWARE OF SAMPLES USED. THEY MAY POSE A BIO HAZARD.
SELECT THE TIME, SPEED AND ACCELERATION FOR THE SHANDON CYTOSPIN PROGRAM
Page 44-47 and page 96
Average cells will require an approximate speed of 1000 rpm with medium acceleration.
Large or fragile cells should be spun at a slower speed (e.g. 500-800 rpm) with low acceleration; small cells or bacteria may require higher speeds (e.g. up to 2000 rpm) with high acceleration. Note that the maximum speed for the EZ Megafunnel is 1500 rpm, and the maximum run duration for EZ Single and Double Cytofunnels is 60 minutes.
RUN THE SHANDON CYTOSPIN
Page 48
When the Shandon Cytospin 4 is programmed and the sealed head is loaded, press START to start the run.
UNLOAD CYTOSPIN SAMPLE CHAMBERS
Page 97
The Shandon Cytospin 4 lid will unlock automatically as soon as the Shandon Cytospin has stopped spinning.
As soon as possible after the Shandon Cytospin has stopped spinning, remove the sealed head from the instrument and open it in a biological safety cabinet. Open the sealed head lid and remove the EZ Cytofunnel assemblies.
FIXATION
Page 98
Fix the samples as soon as possible to avoid autolysis.
STAINING
The specimen can now be stained and examined according to laboratory procedures and examined microscopically.
MICROSCOPY AND DIAGNOSIS
The page and paragraph references refer to the Shandon Cytospin 4 Operator Guide A78310250 issue 4
