Beckman Coulter MicroScan Manuel D'utilisation page 4

3. After the panel has been inoculated with the 50 μL inoculum, lightly tap each side of the panel to help ensure mixing with
the substrate.
NOTE: If the panel is to be read on the WalkAway or autoSCAN-4 the LOC well must also be inoculated.
Incubation
1. Stack panels in groups of 3 to 5 with a cover tray over the top panel to prevent evaporation.
2. Incubate the inoculated panels for 4 hours at 35-37°C in a non-CO
Reading the Panels
1. Remove the Peptidase reagent from the refrigerator at least 30 minutes prior to reading the panels. The Peptidase
reagent may precipitate at 4°C, but it will redissolve at room temperature. Do not expose the Peptidase reagent to
elevated temperatures to accelerate the redissolving of the crystalline precipitate because high temperatures will cause
the reagent to degrade.
2. Read the panels using reflected light from a white background.
3. After 4 hours of incubation, add the Peptidase and Sodium Hydroxide (NaOH) reagents to the appropriate wells.
A. Add 1 drop of Peptidase reagent to the BNAC, HPR, ILE, PRO, TYR, GLY, GGLY, GLAR, GLPR, AARG, LYAL,
ALA, STY and HIS wells. Allow the color to develop for 30 seconds before recording the results. Results must be
recorded within 3 minutes of reagent addition. The presence of ANY shade of pink or red in the solution throughout
the entire well or any portion of the well is interpreted as positive. Colors should be compared to the BNAC well.
Some wells may appear a darker orange than the BNAC well. These should be recorded as negative. Reactions
are positive only if there is pink or red present in the solution. The color in these wells will darken with time. A
negative yellow color may become a darker orange color. This should be interpreted as a negative reaction.
NOTE: Following addition of Peptidase to the BNAC well, it should be yellow. If there is any pink or red present,
the Peptidase reagent should not be used.
B. Add 1 drop of 0.05N NaOH to AGL2, BDF, AGAL, NPC, NAG, CELL and NGAL wells. Wait at least 5 seconds,
but no longer than 5 minutes, before recording results. Any shade of yellow should be recorded as a positive.
Compare the substrate-containing wells to the NPC well.
NOTE: These wells may appear darker than the NPC well. This should be interpreted as negative, but a yellow
color must be present to be considered positive.
4. Two control wells are provided to assist in interpreting the reactions.
A. BNAC = β-naphthylamide control. After addition of Peptidase reagent, this well will be yellow in color.
B. NPC = nitrophenyl control. This well should be colorless.
5. Refer to the RESULTS section for aid in biochemical interpretation.
NOTE: The WalkAway reads wells that do not require reagents first to prevent altered results due to reagent fumes that
may be trapped under the tray lid, therefore, biochemical reactions cannot be verified manually.
RESULTS
Substrate Interpretations
Well Reagent
HPR Add 1 drop of Peptidase reagent. Allow color reaction
to develop for at least 30 seconds, but no longer than
ILE
3 min.
PRO
TYR
GLY
GGLY
GLAR
GLPR
AARG
LYAL
ALA
STY
HIS
SUC1
SUC2
TRE
AGL1 Compare to the NPC control well.
BGL
BGAL
URE
C29879–AB
incubator.
2
Positive
Any shade of
Pink, Red to
Magenta in the
1
solution
Any shade of
Brown or Yellow
Any shade of
2
yellow
Pink to Magenta Clear/Beige to
4 of 141
Negative
Yellow to Orange
Purple
Clear
Yellow