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The present part of the E-sc@n questionnaire is devoted to conventional techniques: how samples are collected, transmitted, received, and treated. A second part will address diagnostic features using conventional stains: how normal, reactive and malignant cells are recognized. A third part will address ancillary techniques, of which immunocytochemistry is the best known, and the most widely applied method.
Your name and degree (optional)First part: Conventional methods
Background – laboratory activity
Your laboratory name and addressYour E-mail address
Number of cases you handle / monthly / annually
Collecting and receiving the fluid
What do you recommend? / 30 ml tubes ---lessmoreSterile containers / ---yesnono particular advice
What sort of container do you receive?
How many tubes?
With EDTA / ---yesno
With another anticoagulant? / ---yesno Which one?
With a fixative? / ---yesno Which one?
When do you process? / Immediately after receiptIn the hour following receiptThe day of receiptUp to 12 hoursUp to 24 hoursUp to 48 hours
Do you incubate at +4°C? / neversomtimes - Why?
Do you freeze? / neversomtimes - Why?
Free text
Medical history and biological data
On receipt, are you aware of the medical history? / ---yesno noteOn receipt, are you aware of the clinical context? / ---yesno note
On receipt, are you aware of radiological data? / ---yesno note
If yes at any step:
By means of a well-designed transmission sheet? / ---yesno noteBy means of handwritten data from the clinician? / ---yesno note
By means of connection with a secured server? / ---yesno note
Other (precise)
On receipt or at any phase of the process, are you aware of the exudative/transudative nature of the effusion? / ---yesno noteFree text
Controls
On receipt, do you note the total quantity of fluid? / ---yesno noteOn receipt, do you note the macroscopic aspect of the fluid? / ---yesno note
Cell fixation
At the laboratory, do you add some fixative? / ---yesno if yes, which one?Free text
Concentrating cells
Before concentrating:
On receipt, do you mix every tube? / ---yesnoDo you shake vigorously (if clot)? / ---yesno
Do you transfer into conical tubes? / ---yesno
Do you add albumine or other coating? / ---yesno
Which coating medium, and what quantity do you use exactly?
Sedimentation:
Do you use sedimentation? / neversometimesoftensistematicallyFor what reasons?
Centrifugation:
Do you use routinely centrifugation? / ---yesno / What speed (or how many g)?What time period? / min / Temperature? / room temperatureat 4°c
If the cell pellet is poorly, or hardly visible, do you use cytocentrifugation? / ---yesno
If no, why?
Cytocentrifugation:
Do you systematically use cytocentrifugation? / ---yesnoDo you know the name and characteristics of your device? / ---yesno note
Do you proceed to a cell count before cytocentrifugation? / ---yesno
Do you use reusable chambers? / ---yesno / Do you use disposable chambers? / ---yesno
If yes, of what type?
What quantity of fluid do you process? / Less than 500 microliter in each chamberMore than 500 microliter in each chamberFree text
Cell blocks not concerned
Are you used to prepare cell blocks? / ---yesnoHow do you proceed?
Shandon Cytoblock method? / Automated method?Personal technique (explain in the free text)
Free text
Liquid based cytology not concerned
Types
Methods
Free text
Removing blood
In case oh haemorrhagic specimen, how do you proceed?
No particular procedure
Wiping of the clot on slides as for a blood film* (if applicable)
*see Spriggs and Boddington, Atlas of serous fluid cytopathology, Kluwer Acad. publishers, 1989, pp. 127-128
Haemolysis:
Haemolysis with ammonium chloride No Yes
Haemolysis with saponine No Yes
Haemolysis with another method No Yes – which one?
Differential centrifugation:
Lymphoprep Ficoll Other medium
Liquid-based cytology:
LBC using haemolytic mediums and filtration (such as Cytyc Thinprep)
LBC and smears together No Yes
Other procedure
Free text
Unstained wet preparations (bright field, or phase-contrast microscopy)
Do you sometimes use unstained preparations? No, never Sometimes
For cholesterol crystals* identification? No Yes
For another purpose? No Yes – Precise
*Are you used to cholesterol crystals appearance on MGG preparations? No Yes
Air-dried direct smears
Are you globally satisfied with your smear technique? No Yes
Do you systematically pour off the supernatant after centrifugation? No Yes
Immediately after centrifugation? Not systematically Yes
Do you keep the tube upside-down when aspirating, as recommended? No Yes
*see Spriggs and Boddington, Atlas of serous fluid cytopathology, Kluwer Acad. publishers, 1989, p. 127
Do you use a micropipette or a glass pipette with a capillary tip?
Do you obtain satisfying smears? No Not always Often Almost always
How many smears do you systematically prepare?
How many of them are reserved for additional techniques?
Is immunocytochemistry performed on smears or on LBC slides?
Never Sometimes Often systematically
Wet-fixed smears
Do you use the same smearing method as for air-dried smears? No Yes
With what fixative? Ethanol Methanol Other
Do you fix immediately (before drying)? No Yes
Staining
Systematically used stains: Haematologic: MGG Giemsa PAS Polychromatic: Papanicolaou
Other:
Do you systematically use MGG or Giemsa + Papanicolaou stains combined? No Yes
Free text
Mounting of slides
Do you systematically mount slides? No Yes
Do you systematically unmount your slides No Yes
If yes, why?
Global appreciation
In your laboratory;
Are you globally satisfied by your techniques? No Yes
Would you consider that you have reached a high quality standard? No Yes
Would you consider that significant improvements have to be done? No Yes
Highest quality domains in your own laboratory: please quote
Controls on receipt
Transmission of medical history and clinical data
Delay before processing
Delay before result
Global processing of fluids before staining
Concentration methods (globally)
Smear technique
Centrifugation and/or cytocentrifugation
Quality of the cell fixation and preservation
Elimination of RBC
Cell blocks
Liquid-based cytology
Stains
Domains which could benefit from improvements;
Controls on receipt
Transmission of medical history and clinical data
Delay before processing
Delay before result
Global processing of fluids before staining
Concentration methods (globally)
Smear technique
Centrifugation and/or cytocentrifugation
Quality of the cell fixation and preservation
Elimination of RBC
Cell blocks
Liquid-based cytology
Stains
Give a subjective, global note to your own preparatory techniques: / 20
Contributors:
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The present questionnaire is intended to provide the most exhaustive view as possible of the way the practicing pathologists deal with effusions. Thank you very much for your help.
Your contribution will be acknowledged and you will be contacted at a later date.
The E-sc@n project team