Transcript Renal cases - Fagdyrlaegen

Cytology cases
Kathleen Tennant BVetMed Cert SAM Cert VC FRCPath
MRCVS
Clinical Lead
Diagnostic Laboratories
Langford Veterinary Services
Aims
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Case based
Mixture of levels
Some recurring problems
Practice ‘screening’ slides
First, the set-up
• A major reason many people won’t screen
their cytology is because they can’t see down
the microscope properly....
Which of these would apply to you?
1. Microscope’s always covered in oil
2. Forgotten what I’m supposed to do with
the condenser
3. Our Diff-Quik’s scummy and won’t stain
right
4. All of the above
5. None of the above
Quick fixes
Microscope’s always covered in oil
Clean it! Ethanol, methanol, not acetone. You can take
the lenses off to make it easier but be careful not to
get anything inside it...
Forgotten what I’m supposed to do with the condenser
Up – just below the stage, if it will move, usually open
Our Diff-Quik’s scummy and won’t stain right
You can scrape the scum off the last one with filter paper
– use small narrow upright jars so you are more
willing to throw it away
Which of the following can not be
successfully used as immersion oil?
1. Liquid paraffin
2. A mix of two different commercial
immersion oils
3. Perfume grade cedar oil
4. Coverslipping glue
Immersion oils
• You cannot mix different oil types – each has
its own refractive index and a combination will
be impossible – clean the oils off both the lens
and the slide to fix it
• Coverslipping glue works suprisingly well for
approximately 20 seconds. Takes about 2
hours to mend the microscope (sorry, Nikon
technician....)
Which of these is not a ‘cytology sin’?
1. Putting thick samples in a slide holder before
they are dry
2. Putting the slides in the same bag as the
histopath sample
3. Staining a slide with Diff-quik for a look and
then sending it to an external lab
4. Staining a slide with Diff-quik then not
including it with the rest of the slides to go
out
Cytology ‘sins’
• Slides going into holders stay wet in the humid
environment for a long time and this can
affect morphology
• Even a small amount of formalin affects the
way the cells stain and drastically cuts down
detail
• External labs prefer to have all slides – the one
that you have stained may be the only one
with material on it
Case 1
• 5 year old Great Dane
from a rescue centre with
cough, pyrexia and
anorexia
• Radiographs show widely
distributed alveolar
pattern
• Tracheal wash taken as
initially unresponsive to
antibiotics
• Gelatinous material
retrieved – direct smears
made
Direct smear, 100x lens oil immersion
– what is this?
Is this/are these....?
1.
2.
3.
4.
5.
6.
Bacteria
Stain precipitate
Necrosis
Inflammation
KY gel
Mucus
KY/ ultrasound gel
• Pink to deeply purple
staining, irregular
flecked material
• Stain precipitate looks
very similar but
(hopefully) should not
be this thick
Mucus
Case 1
• Same dog, second
sample (one reason it’s
good to screen a sample
before waking up – can
hair dry them if in a
hurry, pre- and post
staining)
Tracheal wash # 2
Is this...
1.
2.
3.
4.
Normal
Neutrophilic inflammation
Septic
Mast cell metastasis
Normal tracheal wash
• When screening your samples, consider what
the normal cell population in that area should
be
• Airways have regular columnar epithelial cells
– sometimes the cilia are visible
• Goblet cells (mucus producing) look very
similar to mast cells
Tracheal wash # 2
Same dog, BAL left
Closer up
What’s your diagnosis?
1.
2.
3.
4.
Fungal pneumonia
Gram positive bacterial pneumonia
Gram negative bacterial pneumonia
Mixed bacterial pneumonia
Same dog, BAL left, neutrophils
Septic neutrophilic inflammation
• The demonstration of organisms INSIDE
neutrophils is needed to confirm sepsis
• Use neutrophil size to compare organisms to –
bacteria rarely > 1/20th neutrophil diameter
• Don’t forget that Diff-Quik tells you nothing
about the Gram characteristics of bacteria...
• Neutrophils will often degenerate in the
presence of bacterial toxins, or irritants such
as pancreatic fluid, bile or urine
Non – degenerate/ degenerate
Aspirate from a nodule on a 2 y.o.
DSH’s head
What’s your diagnosis?
1.
2.
3.
4.
5.
Fungal infection
Gram positive bacterial infection
Gram negative bacterial infection
Mycobacterial infection
Mast cell tumour
Aspirate from a nodule on a 2 y.o.
DSH’s head
• The cell here is a
macrophage
• The white (non- staining)
linear stripes within it are
Mycobacteria
• Macrophages are very
good at ‘presenting’ the
cause of some infections
– keep an eye out for
what’s inside them
Peritoneal fluid from an ascitic dog
• The gross appearance
of the fluid is
serosanguinous
• The nucleated cell is a
macrophage
When did the bleeding occur?
1. Current/ iatrogenic
2. Minutes to hours ago
3. Last week
Peritoneal fluid from an ascitic dog
• There are no platelets
present, which makes
current/ iatrogenic bleeding
less likely (not impossible)
• The macrophage has had
time to phagocytose a red
cell – minutes to hours
• Might expect a week old
bleed to have cleared, or
macrophages to contain
haem pigment breakdown
products
Recent haemorrhage - cytology
Long standing haemorrhage - cytology
Longer standing haemorrhage
Tracheal wash from a 7 y.o. Coughing
Yorkshire Terrier
Is this...
1. Normal
2. Squamous cell carcinoma
3. Oropharyngeal contamination and
neutrophilic inflammation
4. Septic neutrophilic inflammation
Oropharyngeal contamination
• Oropharynx contains
nucleated squamous cells
• The ‘striped’ organisms
are Simonsiella, which are
particular to the
oropharynx
• Neutrophilic
inflammation also present
– but interpreting any
culture results will be
difficult
Pollen and mucus
• Other ‘interlopers’ in
airway washes include
plant, food and pollen
material
• Anything green on a
stained slide has to be
self pigmented and is
often of plant origin
Nasal flush from a 3y.o. GSD
Is this...
1.
2.
3.
4.
Bacterial infection
Yeast/fungal infection
Plant contamination
Nematode infection
Yeast infection
• Yeast are generally
larger than bacteria and
often have a nonstaining capsule
• Look for septae
• Aspergillus usually large
and may have long
hyphae
Aspergillus
Yeast from laboratory cactus!
BAL from a 4y.o. Coughing cat
Similar BAL from a coughing dog
Is this...
1.
2.
3.
4.
Eosinophilic inflammation
Basophilic inflammation
Neutrophilic inflammation
Normal
Eosinophils
• Round granules in dogs,
elongated granules
(more difficult to see) in
cats
• Look for potential
causes
• Pulmonary infiltrate
with eosinophils in
dogs, ‘asthma’ in cats
Angiostrongylus
• Note the size of the
organism versus the
dots in the background,
which are neutrophils
• Neutrophilic
inflammation can be
associated with this as
much as eosinophils