Biomerieux Previ Color Gram User Manual

USER’S MANUAL

4501 - 1689 - en

bioMérieux S.A. 69280 Marcy l'Etoile / France		REF	29675
Tel. [phone] - Fax [phone]		version B
http://www.biomerieux.com

IVD

Printed in France / 673 620 399 RCS Lyon	03/2009
© 2008. bioMérieux, S.A.

The performance characteristics and the results of the products presented are subject to a proper and skilled use of the product in strict conformance with the User’s Manual.

Photo and illustration credit: bioMérieux.

bioMérieux, the blue logo and PREVI are used, pending and/or registered trademarks belonging to bioMérieux S.A. or one of its subsidiaries.

Revisions

The list of revisions below summarizes replacements or additional pages in your User’s Manual.

Version	Date of printing	Modifications	Pages modified
A	09/2008	Creation	All
B	03/2009	Addendum # 4501-1842 insert	P 2-6

Table of Contents

1. How to use this manual		1-1
	Warnings ...............................................................................................................................................................	1-1
	Specific warnings...................................................................................................................................................	1-1
2.	General	2-1
	Functional description............................................................................................................................................	2-1
	Key features ..........................................................................................................................................................	2-1
	Intended use..........................................................................................................................................................	2-1
	Specifications ........................................................................................................................................................	2-2
	General ............................................................................................................................................................	2-2
	Performance ....................................................................................................................................................	2-2
	Run timing sequence and reagent consumption - General references ............................................................	2-3
	References ............................................................................................................................................................	2-3
	Description.............................................................................................................................................................	2-4
	Front panel and keypad description .................................................................................................................	2-4
	Back panel .......................................................................................................................................................	2-5
	Bowl .................................................................................................................................................................	2-5
	Maintenance kit................................................................................................................................................	2-5
	Nozzle..............................................................................................................................................................	2-6
	Nozzle cleaning solution ..................................................................................................................................	2-6
	Other parts.......................................................................................................................................................	2-6
3.	Installing the PREVI™Color Gram	3-1
	Unpacking procedure ............................................................................................................................................	3-1
	Installation procedure ............................................................................................................................................	3-2
	Installing waste tubing......................................................................................................................................	3-2
	Connecting power ............................................................................................................................................	3-3
	Installing reagent bottles ..................................................................................................................................	3-4
	Preparation procedure...........................................................................................................................................	3-5
	Nozzle checking...............................................................................................................................................	3-5
	Pattern and volume testing ..............................................................................................................................	3-6
4.	Operating the PREVI™ Color Gram	4-1
	CLEAN reprime stage............................................................................................................................................	4-2
	Pattern test ............................................................................................................................................................	4-3
	Loading slides........................................................................................................................................................	4-5
	Starting a staining cycle.........................................................................................................................................	4-8
	Unloading slides ....................................................................................................................................................	4-9
	CLEAN alcohol purge stage ................................................................................................................................	4-10
	Alcohol fixation (ethanol or methanol) .................................................................................................................	4-11
	Monitoring reagent level ......................................................................................................................................	4-12
	Replacing a reagent bottle .............................................................................................................................	4-12
	Emptying the waste container........................................................................................................................	4-13
	Other settings ......................................................................................................................................................	4-13
	Decolorizer setting .........................................................................................................................................	4-13
	Stain adjustment ............................................................................................................................................	4-15

PREVI™Color Gram User’s Manual

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Table of Contents

5. Maintenance and cleaning	5-1
Preventive maintenance plan ................................................................................................................................	5-1
Preventive maintenance chart..........................................................................................................................	5-1
DAILY maintenance .........................................................................................................................................	5-2
WEEKLY maintenance ....................................................................................................................................	5-2
MONTHLY maintenance..................................................................................................................................	5-3
YEARLY maintenance .....................................................................................................................................	5-3
Nozzle handling .....................................................................................................................................................	5-4
Disassembly.....................................................................................................................................................	5-4
Cleaning...........................................................................................................................................................	5-5
Reassembly .....................................................................................................................................................	5-6
Unclogging the tubing ......................................................................................................................................	5-8
Volume test..........................................................................................................................................................	5-10
B line flushing procedure .....................................................................................................................................	5-13
Priming all lines simultaneously...........................................................................................................................	5-16
Long-term storage ...............................................................................................................................................	5-16
Cleaning ..............................................................................................................................................................	5-18
Cleaning the instrument and the carousel......................................................................................................	5-18
Liquid spills ....................................................................................................................................................	5-18
Preventing injury..................................................................................................................................................	5-18
Disinfecting and/or sterilizing...............................................................................................................................	5-19
Disinfection solutions .....................................................................................................................................	5-19
Disinfection procedure ...................................................................................................................................	5-20
Instrument shipment ............................................................................................................................................	5-22
Disinfection declaration..................................................................................................................................	5-22
Fuse replacement................................................................................................................................................	5-23

6.	Troubleshooting		6-1
	Error messages and Troubleshooting Table..........................................................................................................		6-1
7.	Preventive maintenance chart		7-1
	Note:	Screenshots, figures, and messages are given for information purposes only.

I-2	PREVI™Color Gram User’s Manual

1. How to use this manual

This manual provides the appropriate instructions to install, operate and maintain the

PREVI™ Color Gram instrument.

W a r n i n g s

Different types of warnings are used throughout the manual:

•for safety reasons (DANGER!),

•to ensure that the instruments are maintained in good working condition (CAUTION!),

•for regulatory reasons (WARNING!) or,

•for optimum performance of operations, procedures, etc. (IMPORTANT!).

Before using your instrument, please also read the "General safety and regulatory information" booklet, provided with the instrument, and the above mentioned warnings.

S p e c i f i c w a r n i n g s

DANGER!	The PREVI™ Color Gram must be installed in a well ventilated area.
	If power is lost while the stainer is running, do not open the lid until carousel rotation
	ceases. Check the amber rotation light on the front panel and listen for the sound of
	carousel rotation to cease.
	The anhydrous alcohol used in the instrument is highly flammable. A break or
	malfunction in the reagent delivery system has the potential of releasing up to 500 ml
	of alcohol in and around the instrument. If this occurs, carefully shut off the power to
	the stainer and consult the safety sheet for information in handling alcohol spills.
	Do not use the instrument again until any leaks are repaired.
	Regarding the PREVI™ Color Cytocentrifuge Rotor, lid and rotor gaskets and related
	components are intended to be part of biosafety systems such as are specified in
	international and national biosafety guidelines, and cannot be relied on as the only
	means of safeguarding workers and the environment when handling pathogenic
	micro-organisms.
	bioMérieux recommends distilled water for reagent D and ethanol for reagent E.
	Methanol can also be used for reagent E, nevertheless its use is subject to specific
	safety precautions and requires the instrument to be placed under a safety hood.

CAUTION!	To avoid serious instrument damage, never use reagents other than those supplied by
	bioMérieux.
	Except for both reagent mentioned above, using reagents not supplied by bioMérieux
	may seriously damage the stainer and may void your warranty.

PREVI™Color Gram User’s Manual

1-1

How to use this manual

Specific warnings

IMPORTANT!	Please read this manual carefully before using the PREVI™ Color Gram.
	In all cases of maintenance and repair, work should only be undertaken by a skilled
	technician. Untrained personnel should not attempt to dismantle the instrument.
	Only spare parts supplied or specified by bioMérieux should be used. Fitting of non-
	approved parts may affect the performance and safety features designed into the
	instrument.
	If the equipment is used in a manner not specified in the instructions for use provided
	by bioMérieux, the protection provided by the equipment may be impaired.
	If in doubt, contact your bioMérieux representative.

1-2	PREVI™Color Gram User’s Manual

2. General

F u n c t i o n a l d e s c r i p t i o n

The PREVI™ Color Gram is designed to Gram stain specimen smears on microscope slides.

A Gram stain aids in the presumptive diagnosis of the illness, and it gives preliminary classification of the causative agent.

This microprocessor-controlled slide stainer	is dual purpose. Additionally, the
PREVI™ Color Cytocentrifuge Rotor allows to	quickly sediment sample cells onto
microscope slides for staining or other purposes.

The chemical mechanism of Gram differentiation is based on the differential cell wall permeability to the Crystal Violet-Iodine complex. The Gram stain yields a specimen in which the Gram-positive organisms are purple to black and the Gram-negative organisms are pink to red.

K e y f e a t u r e s

PREVI™ Color Gram’s features are:

•minimized reagent consumption

•fast staining

•high volume staining productivity (12 or 30 slides at a time)

•CLEAN cycle to purge each spray nozzle with ethanol or methanol

•9 operator-selected decolorizer cycles*

•separate reservoir, delivery tube, pump and spray nozzle for each reagent

•operator-selectable ethanol or methanol fixation function to fix samples. The correct accessory must be used for each function.

The PREVI™ Color Cytocentrifuge Rotor is available from bioMérieux as an option offering additional features.

*program no. 2 and 3 recommended by bioMérieux.

I n t e n d e d u s e

	The PREVI™ Color Gram is an in vitro diagnostic medical device for professional use only.
	The PREVI™ Color Gram has been designed to perform automated Gram staining of human
	sample micro-organisms for in vitro diagnostics with bioMérieux reagents only.
	The PREVI™ Color Gram has been qualified with the following cell suspensions:
	•	blood
	•	bronchoalveolar liquid (BAL)
	•	cerebrospinal fluid (CSF)
	•	feces
	•	sputum
	•	vaginal swab
	•	urine.
WARNING!	bioMérieux disclaims any responsibility for the use of the PREVI™ Color Gram with
	biological cell suspensions other than those mentioned above.

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2-1

General

Specifications

S p e c i f i c a t i o n s

General

	PARAMETERS			CHARACTERISTICS
	Main power input			100 – 240 V~ / 50 - 60 Hz
				The power setting is indicated on the back panel.
	Power Consumption			85 W max
	Fuse rating			2A-250V ~ IEC TYPE T
				5x20 mm (0.2x7.9 inches)
	Outside dimensions
	Width			560 mm (22 inches)
	Depth			530 mm (21 inches)
	Height - Lid open			580 mm (23 inches)
	Height - Lid closed			240 mm (9.5 inches)
	Weight			16.8 kg (37 lbs)
	Ambient temperature
	Operation			15 °C to 30 °C (59 °F to 95 °F)
	Storage			- 20 °C to 60 °C (- 4 °F to 140 °F)
	Relative humidity			maximum 80% at up to 31 °C
	Altitude			≤ 2000 m
	Pollution degree		2
	Over-voltage category			II

Performance

	PARAMETERS			CHARACTERISTICS
	Reagent spray nozzles			Each reagent has a separate spray nozzle
				to dispense the correct amount of reagent.
				A - Fuchsin or Safranin (with or without acetone)
	Reagent channel			B - Iodine
				C - Crystal Violet
				D - Distilled water *
				E - Ethanol or methanol **
	Carousel capacity			12 slides
				30 slides
	Run			9 operator-selected decolorizer cycles
				(Program no.2 and no.3 validated by bioMérieux)
	Rotor speeds			20 rpm for staining (rpm - revolutions per minute)
				950 rpm for drying

*Not provided by bioMerieux. Since distilled water and ethanol or methanol are not available from bioMérieux, it must be obtained locally. Water should be filtered (0.2 micron inline filter) to remove bacteria and particulates.

**Ethanol or methanol should be 99.8% pure.

2-2	PREVI™ Color Gram User’s Manual

General

References

Run timing sequence and reagent consumption - General references

The following table includes the general references for a staining cycle at DECOLORIZER 3.

										Slides carousel
	Phases			PUMP			12							30
							Time			Volume			Time			Volume
							sec			ml			sec			ml
	(0)	FIXATION*		(E)			(70)			(3.0)		(82)			(3.5)
	1	CRYSTAL VIOLET		C			35			4.5		43			5.5
	2	WASH		D			25			17.3		34			23.5
	3	IODINE		B			30			4.5		38			5.7
	4	WASH		D			33			22.8		69			47.8
	5	DECOLORIZER		A			39			6.0		47			7.2
	6	WASH		D			20			13.8		34			23.5
	7	HIGH SPEED DRY		-			55			-		55			-
		Total without alcohol fixation					237			69			320			113.3
							4 min						5.3 min
		Total with alcohol fixation					307			144		402			116.8
							5.1 min						6.7 min

*Option for ethanol or methanol fixation.

R e f e r e n c e s

PRODUCTS	REFERENCES

PREVI™Color Gram CD Rom

PREVI™Color Gram User’s Manual

PREVI™Color Cytocentrifuge Rotor User’s Manual

General Safety and Regulatory Information

•fr, en, de, es, it, pt, el, sv, da, pl, hu, sk

•et, lt, lv, ru, ja, tr, na, zh, ko, th

REAGENTS

•Acetone safranin-A

•Safranin-A

•Acetone fuchsin-A

•Fuchsin-A

•Iodine-B

•Crystal violet-C

Nozzles * 2

Nozzle o-ring

Nozzle wire

Nozzle cleaning solution

Slide carousel : 12 / 30

D&E empty bottle

D&E 5-Liter empty bottle

Waste tubing

Waste container

Maintenance kit

See your bioMérieux representative

PREVI™ Color Gram User’s Manual

2-3

General

Description

D e s c r i p t i o n

The PREVI™ Color Gram consists of:

Lid with safety lock

Carousel 30

or

Carousel 12

Bowl (see page 2-5)

Front panel (see page 2-4)

Front panel and keypad description

Indicators

Cytocentrifuge mode selection

Access to PATTERN and VOLUME tests (see "Pattern and volume testing", page 3-6)

Power Rotation

Cleaning process

Right side panel with 5 prime buttons for 5 reagent channels :

Fuchsin or safranin (with or without acetone)

Iodine

Crystal Violet

Distilled water

Ethanol or methanol

Run a staining process

Program the staining setting

Abort any operation or Exit

Digital display

Numeric keys (programs)

Control keys

2-4	PREVI™ Color Gram User’s Manual

General

Description

Back panel

Bowl

DR E C

B

A

DF

Maintenance kit

Volume button:

(signal tone setting)

Exhaust vent

Power switch:

On Off

Fuse door; alternative current

(see "Specifications", page 2-2).

Waste tubing

Power cord

Spray nozzle orifices: DR - Distilled water Rear E - Ethanol or methanol C - Crystal Violet

B - Iodine

A -- Fuchsin or Safranin (with / without acetone)

DF - Distilled water Front

Drive hub

6 nozzle tubes

6 volume test tubes

1 nozzle tool

1 cleaning wire

1 unclogging tool

1 nozzle disassembly tool

2 nozzles

4 nozzle O-ring

Grease (silicon lubricant)

PREVI™ Color Gram User’s Manual

2-5

General

Description

Nozzle

Compression screw

Nozzle housing

Swirl cone

O-ring

Nozzle cleaning solution

	The nozzle cleaning solution is used for the PREVI™ Color Gram maintenance operations.
	The 2.5 L concentrated nozzle cleaning solution (reference 29525) is contained in a 5-liter
	bottle. The concentrate is composed of 25.2 g of Oxalic Acid Dihydrate per liter of deionized
	water.
	It contains at least 1% of Oxalic Acid Dihydrate (the Material Safety Data sheet is available
	on request).
	The nozzle cleaning solution must be reconstituted before use by adding
	2.5 liters of ethanol (C2H6O) or methanol (CH4O) (99.8%) to the 5 L bottle.
	This instruction is symbolized on the bottle label by the following mention:
	“--> + 2.5 L (C2H6O or CH4O)”.
	The reagents required but not provided (ethanol or methanol) may present a chemical risk
	and should be handled with care. Reconstituted Nozzle Cleaning Solution with ethanol or
	methanol, can be hazardous and should be handled with the same care.
	The concentrated nozzle cleaning solution must be stored at 15-25°C until the expiry date
	indicated on the bottle label.
	After reconstitution, the solution must be stored at 15-25°C until the expiry date.
	For practical reasons, the solution can be transferred into a 500 mL or a 1-liter bottle.
	In this case, it is important to record the expiry date on the bottle.
	Please refer to “Maintenance and cleaning” section, page 5-4.
	Waste disposal: the Nozzle Cleaning Solution effluents must be treated in the same
	manner as the reagents, see warning page 5-3.
	Note:	The nozzle cleaning solution is available separately.
		Please contact your bioMérieux representative.
Other parts
	Since the following parts are not available from bioMérieux, they must be obtained locally:
	• ethanol or methanol more than 99.8 % pure
	• distilled water.
	Empty bottles can be provided by bioMérieux (see "References", page 2-3).
WARNING!	ETHANOL OR METHANOL are not provided by bioMérieux.
	The customer should purchase them locally and observe the recommendations, in
	terms of safety and chemical risk.

2-6	PREVI™ Color Gram User’s Manual

3. Installing the PREVI™Color Gram

CAUTION!	If you observe any damage on the packaging or equipment, please contact your
	bioMérieux representative before installing the instrument.
	Place the instrument with at least 30 cm (12 inches) clearance from obstructions
	or hazardous materials.

U n p a c k i n g p r o c e d u r e

Open the 2 boxes and remove the protection (foam packaging).

Check the contents according to the packing lists included in the 2 boxes (instrument and accessories).

Lift the stainer out of the box.

Place the instrument on a flat surface, free from dust and vibration and away from direct sunlight.

PREVI™ Color Gram User’s Manual

3-1

Installing the PREVI™Color Gram

Installation procedure

I n s t a l l a t i o n p r o c e d u r e

CAUTION!

Installing waste tubing

Clip the waste tubing into the PREVI™Color Gram and push until you hear a "click".

Adjust the tube length (L < 2m).

Connect the waste tubing to the waste container.

Drip tray

Install the waste container into a drip tray to prevent from spilling any liquid.

Ensure that the distance between the back panel of the PREVI™Color Gram and the wall is at least 30 cm (12 inches).

Before the PREVI™ Color Gram is installed and switched ON it should be left to stand for at least three hours so that there is no possibility of condensation causing a hazardous situation.

Keep the waste container tube straight and as short as possible: 2 meters (79 inches) maximum.

The waste container tube should be positioned at a lower level than the table.

If possible, bioMérieux recommends you to install the instrument at an extremity of the bench top.

Click

3-2	PREVI™ Color Gram User’s Manual

Installing the PREVI™Color Gram

Installation procedure

Connecting power

Make certain the switch is OFF (O).

Check that the voltage label complies with the power supply.

Insert the electric cord into the power socket.

Plug the power cord into the mains

(AC power).

Turn the power switch ON ( I ).

After 2 seconds the digital display will show:

bioMérieux PREVI Color Gram

Version X.X

X: firmware version.	…	…

PREVI™ Color Gram User’s Manual

3-3

Installing the PREVI™Color Gram

Installation procedure

Installing reagent bottles

DANGER!	Reagents used in the PREVI™ Color Gram contain moderately hazardous
	chemicals that require care in handling. Always use appropriate safety
	measures including gloves and eye protection when handling reagents.

Fill the E bottle with ethanol or methanol 99.8% pure and the D bottle with distilled water.

Place each bottle in the appropriate position.

Open the new bottle, record the letter on the cap and keep it for future use (e.g. long term storage).

Insert the dip tube “E” into the reagent bottle “E” and screw on the ring cap.

Repeat step 3 for bottles D,C,B,A.

Note: For frequent use 5-L bottles D & E are available,

see "References" page 2-3.

To install the bottles, cut the tubing just above the dip tube. Install the bottles in a drip tray.

Right side panel with 5 prime buttons for 5 reagent channels :

Fuchsin or Safranin (with or without acetone)

Iodine

Crystal Violet

Distilled water

Ethanol or methanol

3-4	PREVI™ Color Gram User’s Manual

	Installing the PREVI™Color Gram
	Preparation procedure
P r e p a r a t i o n p r o c e d u r e
IMPORTANT!	To obtain optimum performance, thoroughly purge and prime each reagent
	delivery line using the following instructions.
CAUTION!	This procedure must be applied before the 1st use of the instrument.
WARNING!	Never operate a dry pump for more than 10 seconds.

Nozzle checking

Remove each spray nozzle with the nozzle tool by turning the counter clockwise.

Note the location of each nozzle so you can return it to the original location.

Hold a sheet of white paper in front of

the nozzle A.

Press and hold each prime button.

Stain should appear within 5 seconds.

If this is the case: go to step 6.

If not, immediately stop the procedure and

see "Unclogging the tubing", page 5-8.

PREVI™ Color Gram User’s Manual

3-5

Installing the PREVI™Color Gram

Preparation procedure

Flush with distilled water and wipe dry the bowl and the nozzles.

Replace each spray nozzle by hand and then with the nozzle tool by turning clockwise.

Pattern and volume testing

For first use, please refer to both of the following tests:

•	The "Pattern test", page 4-3

	•	Then the "Volume test", page 5-10.
IMPORTANT!	After this preparation procedure, follow the ROUTINE use described in
	"Operating the PREVI™ Color Gram", page 4-1.

3-6	PREVI™ Color Gram User’s Manual

4. Operating the PREVI™ Color Gram

PREVI™ Color Gram routine process

Key

Orange line:	run process
Blue line:	maintenance process
Grey elements:	optional part

This routine process flow diagram describes the necessary stages for the routine use of the instrument. Details are given in the following sections.

PREVI™ Color Gram User’s Manual

4-1