SYSMEX SP-10 - Staining Troubleshooting Guide; Common Staining Issue Resolution

Document Ref: SUKBMS-24-
601
Version: 2.0
Page | 14
Date: 19/07/2021
Classification
Unrestricted
Whichever combination of stains are used the process must be carried out at the correct pH. If the
pH is too acidic the basophilic components appear too pink. If the pH is too alkaline conversely
there is over staining as too much basic stain is taken up. These reaction are also dependent on
time as DNA binds to staining compounds more rapidly than RNA and Haemoglobin is even slower
and as a results incorrect concentration of stains and/or timings will result in poor stains.
Stain trouble shooting
Appearance
Possible cause
Too blue, nuclei
Eosin concentration too low
Blue to Black
Stock stain exposed to bright day light
Batch of stain solution over used
Staining time too short
Staining solution to acid
Smear to thick
Inadequate time in buffer solution
Too pink
Incorrect proportion of azure B – eosin Y
Buffer pH too low
Excessive washing in buffer solution
Pale staining
Old staining solution
Overused staining solution
Azure A and/or C contamination
High ambient temperature
Neutrophil granules not
stained
Insufficient azure B
Neutrophil granules pseudo-
toxic
Excess azure B
Blue background
Inadequate fixation or prolonged storage before fixation
Blood collection into heparin as anticoagulant