ERG-PIMA-02-05-En How To Interpret Error Codes Using Data From The Pima Analyser

Alere Technologies GmbH
Löbstedter Straße 103-105

07749 Jena,
Pima Error Resolution Guide
Germany
How to interpret Error Codes using data from

the Pima Analyser
Revision date: 5-Nov-12
The information contained in this document is intended to guide Service and Repair activities
For internal use only. Do not hand out to customers.
ERG-PIMA-02-05-EN
How to interpret Error Codes using data from the Pima Analyser
Revision Notification
The following changes have been made compared to the previous version of this document.
Changes to:
Error 810: Misalignment of optics block removed as potential trigger

Error 820, 870: Images and related text for focus beads in the cartridge channel removed
because of change from focus beads to laser spots
Error 825, 830, 840, 850: editorial changes
Error 890: removed because eliminated for all software-versions since 0.38g
P2
Revision date: 5-Nov-12 Alere Technologies GmbH ERG-PIMA-02-05-EN
Accessories required:
To establish a connection with the Pima Analyser you need a PC running Windows XP or higher
and an USB-to-LAN adapter and a Crosswire LAN cable or Molex-to-LAN cable.
Additional documents:
SRG-PIMA-01-XX How to establish a Remote Desktop Connection with the Pima Analyser
SRG-PIMA-02-XX How to download data files from the Pima Analyser
SRG-PIMA-05-XX How to open and close the Pima Analyser
SRG-PIMA-06-XX How to adjust Pima Analyser Optics Block towards the Cartridge Channel
SRG-PIMA-07-XX How to adjust the focus of the Pima Analyser Optics
NOTE
For every Error Code a suggestion is given for the most likely Primary Product that needs to be
named in SFDC when opening a complaint case. Follow these suggestions, but review for
correctness before closing a case in SFDC.
For every Error Code or Error Message a suggestion for a SFDC Trending Code is given.
P3
Contents
Error 810 – Channel Filling (SFDC: 3304) ............................................................................................... 5

Error 820 – Focus Control (SFDC: 3305)................................................................................................. 7
Error 825 – Grid Detection at Focus Control (SFDC: 3305).................................................................. 9
Error 830 – Exposure Time (SFDC: 3306)............................................................................................. 11
Error 840 - Focus Range (SFDC: 3307)................................................................................................. 13
Error 850 – Exposure Position Control (SFDC: 3308) ......................................................................... 15
Error 860 – Reagent Control (SFDC: 3309) ......................................................................................... 17
Error 870 – Correspondence Control (SFDC: 3310) ............................................................................ 19
Error 880 - Cell Movement Control (SFDC: 3311) ................................................................................ 22
Error 910 – Image (SFDC: 3313) ............................................................................................................ 24
Error 920 – Plausibility (SFDC: 3314) ................................................................................................... 25
Error 930 – Homogeneity (SFDC: 3315) ................................................................................................ 26
Error 940 – Gating (SFDC: 3316)............................................................................................................ 27
P4
Error 810 – Channel Filling (SFDC: 3304)

Primary Product: Pima CD4 25X or 100X Cartridge Kit
Control Item:
A change in brightness at the position of the outlet hole of the detection channel (caused by
fluorescent antibody in solution) is checked after each filling attempt. A maximum of 6 filling
attempts are performed before Error 810 is triggered.
Potential Trigger:
• Cartridge cap was not properly closed or failure in the peristaltic loop
• Air bubbles at the outlet hole
Action:
Necessary data can be looked at by connecting the Pima Analyser to your PC

(see SRG-PIMA-01-XX & SRG-PIMA-02-XX).
The result of the control item is recorded in the ControlItems.txt document. In case of an Error 810
the line may read: PrefillingControl delta_perc_90 -1 failed
Images that should be examined can be found in the respective ‘entry’ folder in the ‘.datafifo’ folder
of the Pima Analyser. Important images are CartridgeUsageControl.bmp. and
PrefillingControl_01.bmp to PrefillingControl_06.bmp.
Sequence of images with successful Channel Filling: no Error 810
CartridgeUsageControl.bmp PrefillingControl_02.bmp
Dark circle on slightly gray background Bright background indicates that antibody is
indicates that no antibody is present. present. This background can be so bright that
the whole image appears white.
PrefillingControl_02.bmp indicates that filling
was successful at the second attempt.
No corrective action needed

… continue with following page
P5
Error 810 caused by not properly closed cap or failure in the peristaltic loop
Dark circle on slightly gray background Dark circle on slightly gray background
indicates that no antibody present. indicates that still no antibody present after
6 attempts.
Corrective action: Retraining of site personnel on how to properly close a cartridge
Error 810 caused by air bubbles at the outlet hole
Dark circle on slightly gray background An air bubble prevented the access of blood to
indicates that no antibody present. the outlet hole.
Corrective action: A new cartridge needs to be used. No immediate corrective action by TS
P6
Error 820 – Focus Control (SFDC: 3305)

Primary Product: Pima CD4 25X or 100X Cartridge Kit or Pima Bead Standard
Control Item:
After autofocus of the optics is completed, laser spots in the cartridge channel (red and green
signals) are counted. More than 10 red and green signals need to be detected. If the number is
lower, Error 820 is triggered.
Potential Trigger:
• Number of laser spots in the cartridge is too low
• Dirt on the detection channel lid at the Focus Position
Action:

the line may read: FocusControl bead count 10 failed
The image that should be examined can be found in the respective ‘entry’ folder in the ‘.datafifo’
folder of the Pima Analyser. The important image is FocusControl.bmp.
Image of successful Focus Control: no Error 820
FocusControl.bmp
More than 10 red and green events have been counted.
P7
Error 820 caused by too low number of laser spots in the focus area
FocusControl.bmp
The number of red and green events is too low.
Corrective action: A new cartridge needs to be used. No immediate corrective action by TS.
Error 820 caused by Dirt on the detection channel lid at the Focus Position
FocusControl.bmp
The Analyser set the focus on the fiber. As a
consequence, the number of laser spots required is not
detected.
P8
Error 825 – Grid Detection at Focus Control (SFDC: 3305)

Control Item:
After autofocus of the optics is completed, the Pima Analyser verifies the focus position.
A segmentation image called FocusControl_grid.png of the focus position will be created where a
minimum of 17 signals have to be detected within a 5-by-5 grid. If less than 17 signals are found,
Error 825 is triggered.
NOTE: Error 825 can occur only with Analysers running on software-version 0.39.1 and higher
Potential Trigger:
• Incorrect focus control position
• Misalignment of the of the optics block
Action:

the line may read: grid detection failed
folder of the Pima Analyser. The important image is FocusControl.bmp.
Image of successful Focus Control Grid: no Error 825
FocusControl_Grid.png
5-by-5 grid is detected
... continue with following page
P9
Error 825 caused by low number of laser spots in the focus area
FocusControl.bmp
5-by-5 grid is not detected
P10
Error 830 – Exposure Time (SFDC: 3306)

Control Item:
The exposure settings (time and gain) to acquire cell images are based on the average image
intensity (grayscale 70±5 of 256) for both the red and green filter.
The resulting exposure time must lie between 10 and 2000 milliseconds. In addition, the target
image intensity of 70±5 needs to be reached by changing the exposure time.
Seven attempts are granted for this. If any of these two is not possible, Error 830 is triggered.
NOTE: Pima Analysers up to Pima-D-001472 have target image intensity of 90±5.
Potential Trigger:
• Unusual sample characteristics (like plasma)
• Failure of the LED
Action:

The result of the control item is recorded in the exposure_log.txt documents.
Exposure Time successfully adjusted: no Error 830
exposure_log.txt
Exposure time for both colors was adjusted at an image intensity of 70±5.
P11
Error 830
exposure_log.txt
Exposure time for red was correctly adjusted, but even after 7 attempts the exposure time for
green could not be raised enough to reach an image intensity of 70±5. This triggered Error 830
If the Error is frequent and no sample related issues can be found, contact higher level TS.
P12
Error 840 - Focus Range (SFDC: 3307)

Control Item:
In Pima Analyser running with software version 0.39.1 the distance (z-axis-position) between the
lens and the object (when in focus) needs to lie between 230 incr and 1270 incr. A substraction of
the marker offset is not necessary anymore. If this is not possible, Error 840 is triggered.
NOTE: In Pima Analyser running with software versions lower than 0.39.1 the z-axis-position
AFTER subtraction of the marker offset needs to lie between 230 incr and 1270 incr.
Potential Trigger:
• Dirt on the detection channel lid
• Optics out of focus or defective focus servo
Action:
Necessary data can be looked at by connecting the Pima Analyser to your PC(see SRG-PIMA-01-
XX-EN & SRG-PIMA-02-XX-EN).
The result of the control item is recorded in the ControlItems.txt and autofocus_log.txt files. In case
of an Error 840 the line in ControlItems.txt may read: FocusControl green focus range 210 failed
An example of the autofocus_log.txt file is shown below
Focus between 230 and 1270: no Error 840 Error 840 caused by focus at 210
autofocus_log.txt autofocus_log.txt
No corrective action needed Corrective action: depends on analysis of the

ExposureControl_X_bead_red_X.bmp images
…continue with following page
P13
folder of the Pima Analyser. The important image is ExposureControl_X_bead_red_X.bmp.
(NOTE: this image only exists if there was an Error 840)
Error 840 caused by dirt on the detection channel lid
ExposureControl_X_bead_red_X.bmp.
The Analyser set the focus on the fibers on the detection
channel lid. As a consequence, the distance of the lens
to the channel bottom is outside the acceptable range.
Error 840 caused by optics out of focus
ExposureControl_X_bead_red_X.bmp.
The Analyser could not set the focus within the
acceptable range, because the focus set-points of the
optics are slightly off.
Corrective action: The Analyser optics focus needs to be re-adjusted (see SRG-PIMA-07-XX)
SFDC: 6092 Primary Product: Pima Analyser
P14
Error 850 – Exposure Position Control (SFDC: 3308)

Control Item:
The exposure settings (time and gain) to acquire cell images are based on the average image
intensity (grayscale 70±5 of 256) for both the red and green filter.
In order to do this, a position in the detection channel needs to be validated.
There are a total of 7 predefined image positions, of which at least one position needs to fulfill the
following criteria. Error 850 is triggered if none of the 7 image positions fulfills these criteria.
- Bubble surface less than 2%
- (fluorescent) blood is visible in the image
Potential Trigger:
• Air bubbles in the detection channel
Action:

The result of the control item is recorded in the ControlItems.txt and exposure_log.txt documents.
In case of an Error 850 the line in ControlItems.txt may read: Exposure Control bubble area [%]
65.5 failed
The images that should be examined can be found in the respective ‘entry’ folder in the ‘.datafifo’
folder of the Pima Analyser. The important images are ExposureControl_pos_X.bmp.
Error 850 caused by air bubbles in the detection channel
None of the 7 positions

to adjust exposure time
for cell images met the
acceptance criteria.

P15
Error 850 caused by misalignment of the optics block
ExposureControl_pos_7.bmp
The Analyser cannot adjust exposure time because the
optics block is shifted to the right in relation to the
cartridge detection channel. Bright areas are close to
dark ones. The Analyser does not know what area to set
the exposure time for.
Corrective action: The Analyser optics needs to be re-aligned (see SRG-PIMA-06-XX)

and the optics focus re-adjusted (see SRG-PIMA-07-XX)
SFDC: 6091 Primary Product: Pima Analyser
P16
Error 860 – Reagent Control (SFDC: 3309)

Control Item:
At the end of the experiment it is examined whether at least 21 CD3+CD4+ cells can be detected
at the Reagent Control position. If this is not possible, Error 860 is triggered.
Potential Trigger:
• Defective Reagent Control (many reasons)
• Air bubbles at the Reagent Control position
Action:

the line may read: PositiveControl cell count post/pre 9/6 failed
of the Analyser. Important images are PositiveControl_2_0.bmp and PositiveControl_2_1.bmp
Image of successful Reagent Control: no error 860
PositiveControl_2_0.bmp
More than 21 CD3+CD4+ cells can be detected at the
Reagent Control position.
Error 860 caused by Defective Reagent Control
PositiveControl_2_0.bmp
For whatever reason, less 21 CD3+CD4+ cells are
detected at the Reagent Control position.
Corrective action: A new cartridge needs to be used.

No immediate corrective action by TS
P17
Error 860 caused by air bubbles at the Reagent Control position
PositiveControl_2_0.bmp PositiveControl_2_1.bmp
P18
Error 870 – Correspondence Control (SFDC: 3310)

Control Item:
The laser spots at the focus position emit (after excitation) light in a broad range of wavelengths.
This enables their detection with both the red and green filter of the Pima Analyser. Due to the
chromatic aberration (refraction of the light by the optical lens, depending on the wavelength) the
red and/or green signals of the laser spots are not located at the same position in the image. This
misalignment is dependent on the optical lens and the filter lenses. During manufacturing the
extent of misalignment for every position on the image is determined and the parameters are
integrated in the image analysis algorithm. A max shift of 6 pixels is allowed between the images.
If the shift is larger, Error 870 is triggered.
IMPORTANT: It is therefore not possible to switch optic blocks from one instrument to the other.
Potential Trigger:
• Damaged optics block
• Defective filter servo
Action:

the line may read: CorrespondenceControl shift 7.1 failed
The images that should be examined can be found in the respective ‘entry’ folder in the ‘.datafifo’
folder of the Pima Analyser. The important images are FocusControl.bmp and
CorrespondenceControl.png.
P19
Sequence of images with successful Correspondence Control: no Error 870
FocusControl.bmp CorrespondenceControl.png
Error 870 caused by low number of laser spots in the focus area
The number of red and green events is too low and the events are inhomogeneously distributed
so that the Pima Analyser has difficulties aligning corresponding signals. In cases of images that
are extremely out of focus, Error 870 may also be triggered.
If the Error is frequent and no cartridge related issues can be found, contact higher level TS.
P20
Error 870 caused by defective filter servo
All events in FocusControl.bmp appear yellow because the corresponding red and green events
are located at the same positions across the whole image. Since a shift is expected in certain
areas of the image (defined during manufacturing), Error 870 will be triggered.
Corrective action: Contact higher level TS. SFDC: 6034 Primary Product: Pima Analyser
P21
Error 880 - Cell Movement Control (SFDC: 3311)

Control Item:
Right before and immediately after the capture of the 20 images for the determination of the CD4
cell count, cell movement control images are taken to check whether the blood has moved in the
detection channel during the analysis. It is paramount that the blood has properly settled in the
channel, otherwise moving cells may impede on the ability of the Analyser to count them.
Two successive images are taken at the same position with a short time delay using the same filter
setting. After overlapping the images the distance of the cells is evaluated (this is the distance the
cells have moved in the blood during the time between the two images). A maximum shift of 8 pixel
is allowed between the images. If the movement is larger, Error 880 is triggered.
Potential Trigger:
• Cells have not settled completely before image taking (can be caused by air bubbles in the
detection channel, slow sedimentation rate with some blood samples or vibrating surfaces)
Action:

the line may read: MovementControl shift 9.9 failed
of the Pima Analyser. Important images are MovementControl_1.bmp and
MovementControl_2.bmp.
Sequence of images with successful Movement Control: no Error 880
MovementControl_1.bmp MovementControl_2.bmp
In both images (before and after the analysis) all signals appear yellow. This means the signals
from both images are on top of each other. No cell movement.

P22
MovementControl_1.bmp MovementControl_2.bmp
The air bubble in the movement control area was heated up by the excitation beam of the LED
when the first image (here colored in red) was taken. The expanding air pushed the cells out of
position before the second image (here colored in green) was taken. The two signals do not
overlap, triggering Error 880.
NOTE: The error can also be triggered before the actual analysis for CD4 count started. In this
case no MovementControl_2.bmp was taken.
P23
Error 910 – Image (SFDC: 3313)

Control Item:
The Pima Analyser uses 20 images to determine the absolute CD4 cell count. At least 11 of these
images need to be valid. An image can be invalidated due to several reasons, mainly air bubbles.
If there are less than 11 valid images available, Error 910 is triggered.
Potential Trigger:
• Air bubbles in the detection channel
Action:

of the Pima Analyser. Important images are XX.bmp.
The amount of air bubbles in the detection channel invalidated too many images to give a valid
result.
P24
Error 920 – Plausibility (SFDC: 3314)

Control Item:
Error 920 occurs because less CD3 carrying cells (i.e. lymphocytes; ‘CD3+CD4+’ [redgreen] plus
'CD3+CD4-' [red]) were counted than monocytes (‘CD3-CD4+’ [green]). This is a biologically
implausible result and therefore invalid.
Potential Trigger:
• cause is unknown, but in general sample related
Action:

The result of the control item is recorded in the PostProcess_CellcountLog.txt document.
Plausibility successfully passed: no Error 920
PostProcess_CellcountLog.txt
The sum of ‘CountRed’ and ‘CountRedGreen’ is larger than ‘CountGreen‘. The result is
biologically plausible
Error 920
PostProcess_CellcountLog.txt
The sum of ‘CountRed’ and ‘CountRedGreen’ is smaller than ‘CountGreen‘. The result is
biologically implausible. Error 920 is triggered.
P25
Error 930 – Homogeneity (SFDC: 3315)

Control Item:
When filling the cartridge with blood the lyophilized reagents (including the CD3 and CD4
antibodies marked with fluorescent dyes) contained in the cartridge capillary get re-suspended.
The reagents are distributed in the blood by diffusion and the mixing process. However the mixing
may not always be perfect, causing inhomogeneous distribution of fluorescent signal along the
detection channel.
The gradient is computed between the darkest successive pictures as difference of the average
values of the 5 brightest successive pictures. If it exceeds a grey value of 80 the Error 930 is
triggered.
Potential Trigger:
• Fluorescent antibodies are inhomogeneously distributed in the detection channel.
Action:

(see SRG-PIMA-01-XX-EN & SRG-PIMA-02-XX-EN).
of the Pima Analyser. Important images are XX.bmp.
Error 930 caused by inhomogeneous distribution of fluorescent antibodies
XX.bmp
A gradient of fluorescent signal is visible along the detection channel. Error 930 is triggered.
P26
Error 940 – Gating (SFDC: 3316)

Control Item:
The Gating algorithm examines whether there are more than one distinct population of objects. If,
based on the detected contrasts, two or more populations can be separated. A gate can be set
between them (hence, gating). Since additional gating is not intended in our algorithm, Error 940 is
triggered.
Potential Trigger:
• cause is unknown, but in general sample related
• fluorescing dust particles in the detection channel that produce their own population in the
scatter plot
Action:

folder of the Pima Analyser. The important image is PostProcess_Scatterplot.png.
No additional gating: no Error 940
PostProcess_Scatterplot.png
A single population of cells is
identified between the x and y axis.
P27
Error 940
PostProcess_Scatterplot.png
Two populations of cells are identified
between the x and y axis.
P28

ERG-PIMA-02-05-En How To Interpret Error Codes Using Data From The Pima Analyser

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Alere Technologies GmbH

Löbstedter Straße 103-105

How to interpret Error Codes using data from

Revision date: 5-Nov-12

For internal use only. Do not hand out to customers.

Error 810: Misalignment of optics block removed as potential trigger

SRG-PIMA-02-XX How to download data files from the Pima Analyser

SRG-PIMA-05-XX How to open and close the Pima Analyser

SRG-PIMA-07-XX How to adjust the focus of the Pima Analyser Optics

Error 810 – Channel Filling (SFDC: 3304) ............................................................................................... 5

Error 810 – Channel Filling (SFDC: 3304)

• Air bubbles at the outlet hole

Necessary data can be looked at by connecting the Pima Analyser to your PC

Sequence of images with successful Channel Filling: no Error 810

No corrective action needed

Corrective action: Retraining of site personnel on how to properly close a cartridge

Error 810 caused by air bubbles at the outlet hole

Corrective action: A new cartridge needs to be used. No immediate corrective action by TS

Error 820 – Focus Control (SFDC: 3305)

• Number of laser spots in the cartridge is too low

• Dirt on the detection channel lid at the Focus Position

Necessary data can be looked at by connecting the Pima Analyser to your PC

Image of successful Focus Control: no Error 820

No corrective action needed

… continue with following page

Error 825 – Grid Detection at Focus Control (SFDC: 3305)

• Number of laser spots in the cartridge is too low

• Incorrect focus control position

• Misalignment of the of the optics block

Necessary data can be looked at by connecting the Pima Analyser to your PC

Image of successful Focus Control Grid: no Error 825

No corrective action needed

... continue with following page

Error 830 – Exposure Time (SFDC: 3306)

NOTE: Pima Analysers up to Pima-D-001472 have target image intensity of 90±5.

• Unusual sample characteristics (like plasma)

• Failure of the LED

Necessary data can be looked at by connecting the Pima Analyser to your PC

Exposure Time successfully adjusted: no Error 830

No corrective action needed

… continue with following page

Error 840 - Focus Range (SFDC: 3307)

• Dirt on the detection channel lid

• Optics out of focus or defective focus servo

No corrective action needed Corrective action: depends on analysis of the

Error 840 caused by dirt on the detection channel lid

Error 840 caused by optics out of focus

SFDC: 6092 Primary Product: Pima Analyser

Error 850 – Exposure Position Control (SFDC: 3308)

- Bubble surface less than 2%

- (fluorescent) blood is visible in the image

• Air bubbles in the detection channel

• Misalignment of the of the optics block

Necessary data can be looked at by connecting the Pima Analyser to your PC

Error 850 caused by air bubbles in the detection channel

None of the 7 positions

Corrective action: A new cartridge needs to be used. No immediate corrective action by TS

Error 850 caused by misalignment of the optics block

Corrective action: The Analyser optics needs to be re-aligned (see SRG-PIMA-06-XX)

Error 860 – Reagent Control (SFDC: 3309)

• Defective Reagent Control (many reasons)

• Air bubbles at the Reagent Control position

Necessary data can be looked at by connecting the Pima Analyser to your PC