GuangzhouWanfu BiotechnologyCo., Ltd.

Number：VS—VM/RD—1—1211—00

Fully Automated Immunohistochemistry Staining System (PA-3600)

Packaging and TransportationTestingReport

SolutionDevelopment

Hardware and New Technology Department/Prepared by: Date:

SchemeAudit

Hardware and New Technology Department/ReviewPerson: Date:

R&D Department 1/Reviewer: Date:

Scheme Approved

R&DLeader: Date:

Table of Contents

Directory 1

Chapter 1: Overview 2

1.1 Verification Purpose 2

1.2 Responsibilities 2

1.3 Project Basic Information 2

1.4 References 2

Chapter 2 Test Plan 3

2.1 Overall Performance Indicators 3

2.1.1 Appearance and Structure Verification 3

2.1.2 Instrument Function Verification 3

2.1.3 Basic performance verification 3

2.1.4 Test vibration, collision, simulated transportation and drop test 4

2.1.5 Transportation Verification 4

2.4 Overall Machine Performance Verification Plan 5

2.4.1 Appearance and Structure 5

2.4.2 Instrument Functional Test 5

2.4.3 Basic Performance Testing 6

2.4.4 Test vibration, collision, simulated transportation and drop test 9

2.4.5 Transport Verification 11

Chapter 3 Verification Records 12

3.1 Verification Result 12

3.2 Test Data Records 12

3.2.1Performance test data before and after vibration, collision, simulated transportation and drop tests/ 12

3.2.2 Performance test data before/after transportation test 18

Chapter 1: Overview

1.1 Verification Purpose

To ensurethe fully automated immunohistochemical staining machinePA-3600prototypedevelopmentprototype can achieve the required stability, refer to the " GB/T 14710-2009Medical electrical equipment environmental requirements and test methods" climate environmental test Igroup requirements, " Fully automated immunohistochemical staining machinePA-3600technical requirements" and " ISTA 1B—2005 Packaging product test standards" for the developmentprototype to conduct packaging and transportation verification.

1.2 Responsibilities

（1）Hardware and New Technology DepartmentSystem Testing Group:Responsible forthe design and implementation ofthe test plan, andconfirming the test results andcompleting the test report；

(2)InstrumentR&D Department 1 :Responsible for providing the test prototype and test software for this test..

1.3 Project Basic Information

1.3.1 Instrument Use Environment

1）Working temperature: 18℃～30 ℃；

2）Relative humidity:20%～70%（No condensation）；

3）Atmospheric pressure:Less than2000m；

4）Rated voltage:AC100V～240V；

5） Frequency:50Hz/60Hz；

6）Avoid direct exposure to strong light;

7）Use in a well-grounded environment and avoid dust, acidic volatile gases, vibration and strong electromagnetic interference.

1.3.2 Test prototype

Prototype numbers are:PA36002009100001(abbreviated as1#)

1.4 References

(1 ) PA-3600 Fully Automatic Immunohistochemical Staining Instrument Technical Requirements ;

(2)《GB/T 14710-2009 Environmental requirements and test methods for medical electrical equipment》

(3) ISTA 1B—2005 Packaging Product Test Standards

Chapter 2 Test Plan

2.1 Overall Performance Indicators

2.1.1 Appearance and StructureVerification

1) The appearance should be clean, without cracks, burrs or scratches, and the text and markings on the panel should be clear and accurate;

2) Fully automated immunohistochemical stainingmachinemovementparts should be smooth, without jamming or jumping, operate smoothly and without significant noise;

3) Fastener links should be firm and reliable, and no looseness is allowed.

2.1.2 Instrument FunctionVerification

1）Used for pre-pathological analysis sample processing, fully automated completion including dewaxing, antigen retrieval, staining, and fluorescence in situ hybridization (FISH) detection pretreatment, post-hybridization washing；

2）The fully automatic immunohistochemical staining machine has a total of36staining positions；

3）The fully automated immunohistochemical staining machine has40reagent positions；

4）Equippedwith a slide QR codescanning system；

5）The software interface can display the buffer volume in real time；

6）Has the function of classifying and storing waste liquid；

7）Possesses a patient information management system；

8）Has a reagent information management system

2.1.3 Basic Performance Verification

2.1.3.1 Dyeingmodule heating componenttemperatureaccuracy

The dyeing module requires the following temperature control points:Tat37℃,65℃,85℃,101℃:TheglassNTCtemperature measurement for all the collection points in the reaction area temperature control point (the maximum value of the stable temperature (MAX) and the minimum value (MIN)) should be within the range ofT±2℃.

2.1.3.2 Dyeing Module Heating ComponentTemperatureFluctuation

Table2.3.1.2.1 Temperature Fluctuation Determination Standards

2.1.3.3 Reagent Addition Accuracy

Reagent addition volume at100μL、130μL、240μL、370μL、500μLthese control points require: P_vdeviation ≤3%。

2.1.3.4 Reagent addition volume repeatability

Reagent addition volumes at100μL、130μL、240μL、370μL、500μLThese control points require:CV≤5%。

2.1.4 VibrationImpact、Simulated Transportand DropTest

After the testhost and packageappearance inspection;

1) External packaging conditionNo damageand sample machineShould be neat and clean, free of cracks, burrs or scratches, the text and logos on the panel should be clear and accurate;

2) Fully automated immunohistochemical stainingmachine movementparts should be smooth, without jamming or jumping, operate smoothly and without significant noise;

3) Fastener connections should be secure and reliable, and should not be loose.

Instrument function test after trial:

1）For sample processing prior to pathological analysis, fully automated completion of dewaxing, antigen retrieval, staining and fluorescence in situ hybridization (FISH）detection preprocessing, post-hybridization washing；

2）The fully automatic immunohistochemical staining machine has36staining sites；

3）The fully automated immunohistochemical staining machine has40reagent positions;

4）Equippedwith slide QR codescanning system；

5）The software interface can display the buffer volume in real time；

6）Features liquid waste classification storage function；

7）Has a patient information management system；

8）Has a reagent information management system

Instrument Performance Testing after Trials

1）The temperature control range of the dyeing module heating component is:37℃、65℃、85℃、101℃。

Temperature accuracy is ±2℃, temperature fluctuation is less than 0.5℃.

2）Reagent addition amount

Reagent addition volume control range is:100μL、130μL、240μL、370μL、500μL。

Dosage accuracy ≤3%, coefficient of variation CV≤5%.

2.1.5 Transportation Verification

After the experimenthost and packageappearance inspection;

1) Outer packaging conditionNo damageand sample machineshould be clean, without cracks, burrs or scratches, the text and logos on the panel should be clear and accurate;

2) Fully automated immunohistochemical staining machine movement parts should be smooth, without jamming or skipping, operate smoothly and without significant noise;

3) Fastener connections shall be secure and reliable, without looseness.

Instrument function testing after trial:

1）Used for sample processing before pathological analysis, fully automated completion of dewaxing, antigen retrieval, staining and fluorescence in situ hybridization (FISH ) detection pretreatment, post-hybridization washing；

2）The fully automated immunohistochemical staining machine has36staining positions；

3）The fully automatic immunohistochemical staining machine has40reagent positions；

4）Equipped withslide QR codescanning system;

5）The software interface can display the buffer solution volume in real time；

6）It has the function of classifying and storing waste liquid；

7）Have a patient information management system；

8）Has a reagent information management system

Instrument Performance Testing After Experimentation

1）The temperature control range of the dyeing module heating component is:37℃、65℃、85℃、101℃.

Temperature accuracy is ±2℃, temperature fluctuation is less than 0.5℃.

2）Reagent addition amount

The reagent addition volume control range is:100μL、130μL、240μL、370μL、500μL。

Accuracy of dosage≤3%, coefficient of variationCV≤5%.

2.4 Whole Machine Performance VerificationScheme

2.4.1 Appearance and Structure

2.4.1.1 Test method

Inspect1prototype,visually inspect the appearance, structure, buttons, and fastener connections of the prototype in natural light with normal or corrected vision.

2.4.1.2 Judgment Criteria

1) The appearance should be clean, free of cracks, burrs or scratches, and the text and markings on the panel should be clear and accurate;

2) Fully automated immunohistochemical stainingmachinemotionparts should be smooth, without jamming or jumping, working smoothly and without significant noise;

3) Fastener links should be firm and reliable and should not be loose.

2.4.2 Instrument Function Test

2.4.2.1 Test methods

Check1prototypeand system software functions, according tothe followingrequirements for standardized operationconduct full-process testing:

Visually assess whether the fully automatic immunohistochemical staining machine has36positionsfulfillingthe design requirements.

Visually inspect and manually install the reagent rack, confirm the number of reagent bottles that can be installed ;

Is there aQR code scansystem;

Visually inspect the software interface buffer bottle options;

Visually assess if there are two separate categories for storing waste liquid containers;

Detect if the software interface has a patient information management system ;

Detect whether the software interface has a reagent information management system;

2.4.2.2 Judgment Criteria

1）For sample processing before pathological analysis, it automatically completes the processes including dewaxing, antigen retrieval, staining, and pre-processing for fluorescence in situ hybridization (FISH ) detection, as well as post-hybridization washing ;

2）The fully automated immunohistochemical staining machine has36staining sites；

3）The fully automated immunohistochemical staining machine has40reagent positions；

4）Equippedwith slide QR codescanning system；

5）The software interface can display the buffer volume in real time；

6）Has the function of classifying and storing waste liquid；

7）Has a patient information management system；

8）Has a reagent information management system.

2.4.3 Basic performance testing

2.4.3.1 Dyeing module heating component temperature accuracy

2.4.3.1.1 Test method

Use a temperature-measuring slide (with a temperature probe attached) to test according to the following method:

Lowermachine control36modules together, heat them all, prepare the test slides before testing, fix the working area of the slide1个NTCtemperature sensor,NTCas close to the upper surface as possible to ensure the flatness of the slide surface;

Prototypepower on, place theglassslide and control the cover according to table3.3.1.1press down to the preset position and then reset, then select the different temperature points for different testscover pressing preset position， control the heating element to reach the temperature control point corresponding to the preset positionT；

③ After waiting for the temperature to stabilize (please refer to the temperature stabilization time table for the stabilization time at each temperature point), the sampling interval is1S,the total samplingtime is5min. Select the maximum (MAX) and minimum (MIN) temperatures within the stable temperature range.

Table2.4.3.1.1 Correspondence Table of Preset Locations and Temperature Control Points:

2.4.3.1.2Judgment criteria

In the dyeing module, at temperature control pointsTof37℃, 65℃, 85℃, 101℃, respectively, the following requirements are made: temperature measurement of the NTC temperature sensor at the temperature control point in the reaction zone, all collection points (maximum temperature (MAX）and minimum temperature (MIN)) should be within the range of T±2℃.

2.4.3.2 Dyeing Module Heating ComponentTemperatureFluctuation

2.4.3.2.1 Test Method

Test using a temperature measurement slide (with a temperature probe attached) in accordance with the following method:

① Lowermachine controlall modules are heated together. Prepare the test slides before testing, and the NTC temperature sensor in the working area of the slide is fixed. NTCtemperature sensor, NTCas close as possible to the upper surface to ensure that the surface of the slide is flat. SampleTurn on the power, place the slideand control the cover slip according to Table 2.4.3.1.1press down to the preset position and reset. Then, according to different test temperature points, select the cover slip pressing down preset position, control the heating plate to reach the temperature control point corresponding to the preset position T;

② Wait for the temperature to stabilize (please refer to the temperature stabilization time table for the stabilization time at each temperature point), then collect the temperature. The collection interval is 1S, the total collection time is 5min. Calculate the temperature fluctuation according to the formula (2), and record the temperature data.

Related Calculation Formulas:

△Tf = ( Tf_max - Tf_min ) / 2………………………………（2）

Where:

△Tf ------ Temperature fluctuation, unit is ℃;

Tf_max ------ Test point is stable5minwithin the measured highest temperature value, the unit is℃；

Tf_min ------ Testing point is stable5minwithin the measured minimum temperature value, the unit is ℃.

2.4.3.2.2Judgment Criteria

Table2.4.3.2.1 Criteria for Determining Temperature Fluctuation

2.4.3.3 Reagent addition volume accuracy

2.4.3.3.1 Test method

（1）Weigh multiple centrifuge tubes with pre-prepared labels using a high-precision balance (accuracy to 0.0001g) before the sample is added, record the weight as M₁. After the pump is initialized, measure the density of deionized water ρ_{deionized water} using a densitometer. Then, inject the controlled amount of deionized water into the centrifuge tube and record the weight as M2. _{Compare the mass of the centrifuge tube before and after adding the liquid according to the controlled points of the reagent dosage:} 100μL, 130μL, 240μL, 370μL, 500μL. Repeat the measurement 5 times.

（2）Letthecontrol_Rvariable beV，calculate the actualsamplingamountViaccording to formula (2).Calculate the percentage deviation ofdispensingP_Vaccording to formula (3).

Related Calculation Formulas:

V = ………………………………………（2）

P_v = ……………………………（3）

Where:

We -----------------actualsample volume(i=1,2,3,4,5);

V_R ----------------Control quantity；

M₁ ----------------Centrifuge before adding sampleTube weight；

M₂ ----------------Weight of centrifuge tube after adding sample；

ρ_{Deionized water}------------Deionized water density (measured using a hydrometer);

P_v -----------------Spit sample deviationpercentage.

2.4.3.3.3 Judgment criteria

Reagent addition volume at100μL、130μL、240μL、370μL、500μLThese control points require: P_vdeviation ≤3%。

2.4.3.4 Reagent addition volume repeatability

2.4.3.4.1 Test method

（1）Weigh multiple labeled centrifuge tubes using a high-precision balance (precision of0.0001g) to determine their initial weights. After controlling the pump initialization,use a densitometer to measure the density of deionized waterρ_{deionized water}. Theninject the above-controlled amount of deionized water into the centrifuge tubes. Based on the control points for reagent addition:100μL，130μL， 240μL，370μL、500μLcompare the mass of the centrifuge tube before and after adding the liquid. Repeat the measurement5times.

（2）Let the control _R be the control volume， calculate the actual addition volume Vi according to formula (2). Calculate the actual addition volume average valueV_AVE according to formula (4). Calculate the standard deviation of the actual addition volume VSTD according to formula (₅). Calculate the coefficient of variation CV according to formula (6).

Related Calculation Formulas:

V_AVE = ………………………（4）

V_std = …………………（5）

CV = ……………………………（6）

Where:

We -----------------actualsample volume(i=1,2,3,4,5)；

V_AVE --------------Actualaverage sample volume；

V_STD --------------Actualsample volume standard deviation；

CV ----------------Coefficient of variation.

2.4.3.4.2 Judgment Criteria

Reagent addition volumes at100μL、130μL、240μL、370μL、500μLthese control points require:CV≤5%。

2.4.4 Vibration, shock, simulated transport and droptests

2.4.4.1 Preparation before Testing

1、Test1instrument, instrument temperature is stable at normal room temperature (initial detection):

A、Checktheouter packaging andsample machine check the appearance and structure of the sample machine;

B、Checktheprototypeand system software functionality, conduct full process testing;

C、Check all peripheral interfaces;

D、Control points for each test using deionized water with 5 reagent dosages: 100μL、130μL、240μL、370μL、500μL；

E、AccordingtoTable3.3.1.1.1, test36modulesmodulesatallheatedtemperatures:37℃、65℃、85℃and101℃ and record the temperatures;

2、After recording the status, pack the instrument according to the actual packaging.

2.4.4.2 Vibration, collision, simulated transportation vibration and simulated transportation drop sequence testing

1）VibrationTest Method:

1）The sample device is placed on the vibration table, and the vibration equipment parameters are set as follows:① Frequency range:5~20~5Hz；② Amplitude value:0.15mm；③ Sweep frequency cycles:10 times; ④ Sweep rate: ≤1 octave/minute；Vibration tests are performed in the three directions of the X/Y/Z axes.

2）Inspect the packaging and appearance of the sample for damage after the test without changing the packaging and sample location：（Proceed directly to the collision test after inspection).

2） CollisionTest

1）After the sample device is packaged, it is fixed on the collision equipment, and the collision equipment parameters are set as follows: ① Acceleration:10m/s²；② Pulse duration:11±2ms；③ Number of collisions:500±10times；③ Pulse repetition rate:1.0Hz；④ Pulse waveform: half sine wave；Collision tests are conducted in sixdirections.

3）Simulation transportation vibration testmethod

Test speed180CPM, frequency3.0HZ,Test duration2H(intensified), After the test, inspect the packaging and the appearance of the sample for damage without changing the packaging and sample placement; After the inspection is complete, proceed to the drop test.

4）Drop TestMethod

6sides are subjected to free fall, the falling height150mm.After the test is completed, the following tests are carried out:

A、Check the prototype; check the prototype appearance and structure;

B、Checktheprototypeand system software functions, conduct full process testing;

C、Check all external device interfaces;

D、Control points using deionized water for each test5timeswith reagent dosage added:100μL、130μL、240μL、370μL、500μL；

E、AccordingtoTable.2.4.3.1.1.1,thetemperatureof36moduleswastestedatallheatingconditions:37℃、65℃、85℃and101℃.Theresultswererecorded.

2.4.4.3 Testing/Post-Check

1) Inspect the condition of the outer packaging during vibration, collision, simulated transport vibration, and drop tests

2）Perform the following checks after all experiments are completed:

A、Checkthe condition of the outer packaging/Sample inspection Appearance, structure of the sample machine;

B、Checktheprototypeand system software functions, conduct full-process testing;

C、Check each peripheral interface;

D、Control points for reagent dosage using deionized water measured 5 times:100μL、130μL、240μL、370μL、500μL；

E、AccordingtoTable2.4.3.1.1,36modulesweretestedatallheatedtemperaturepoints:37℃、65℃、85℃and101℃andthetemperatureswererecorded.

2.4.4.4 Judgment Criteria

Based on theISTA 1B—2005 Packaging Product Test Standards,GB/T 14710-2009 Environmental Requirements and Test Methods for Medical Devices,Fully Automatic Immunohistochemical Staining MachinePA-3600Technical Requirements”, sequential tests were conducted on the prototype: vibration test, collision test, simulated transportation testand drop test

After the testhost and packageappearance inspection;

1) External packaging conditionNo damageand sample machineshould be neat and tidy, without cracks, burrs or scratches. The text and logos on the panel should be clear and accurate.

2) Fully automatic immunohistochemical stainingmachine movementparts should be smooth and free of sticking or jumping, work smoothly and without significant noise;

3) Fastener connections must be strong and reliable, and must not be loose.

Post-experiment Instrument Functionality Test

1）Used for sample processing before pathological analysis, fully automated completion including dewaxing, antigen retrieval, staining and fluorescence in situ hybridization (FISH ) detection pretreatment, post-hybridization washing ;

2）The fully automated immunohistochemical staining machine has36staining positions；

3）The fully automated immunohistochemical staining machine has40reagent positions；

4）Equippedwithslide QR codescanning system;

5）The software interface can display the buffer volume in real time；

6）Features for classified storage of waste liquid；

7）Has a patient information management system；

8）Has reagent information management system

Instrument performance testing after the experiment:

1）The temperature control range of the dyeing module heating component is:37℃、65℃、85℃、101℃。

Temperature accuracy is ±2℃, and the temperature fluctuation is less than 0.5℃.

2）Reagent addition amount

Reagent addition volume control range is:100μL、130μL、240μL、370μL、500μL。

Accuracy of dosage addition ≤3%, coefficient of variationCV≤5%.

2.4.5 Transportation Verification

2.4.5.1 Test Methods

1、Test1instrument, instrument temperature is stable at normal room temperature (initial detection):

A、Inspectthe outer packaging and sample machine, check the appearance and structure of the sample machine;

B、Checktheprototypeand system software functions, conduct full process testing;

C,Check each peripheral interface;

D、Control points for testing reagent dosage with deionized water5times: 100μL、130μL、240μL、370μL、500μL；

E、按表2.4.3.1.1.1分别测试36个模组模组在全部加热状态下控温点：37℃、65℃、85℃和101℃的温度并记录；

2、After recording the status, pack the instrument according to the actual packaging.

<一> Transportation test:

1）The equipment is tied to the rear of the truck with the label “upwards”;

2.）During the experiment, the load of the car should be ensured to be1/3of the rated load.

3）Road surface: According toJTG B01-2003standards for Class III highway;

4）Driving distance:200km；

5）Driving speed:30km/h~40km/h。

2.4.5.2 Judgment Criteria

After testingthe host and packagingappearance inspection;

1) External packaging conditionNo damageand sample machineShould be clean, no cracks, burrs or scratches, the text and markings on the panel should be clear and accurate;

2) Fully automated immunohistochemical stainingmachine movementparts should be smooth, without jamming or jumping, work smoothly and without significant noise;

3) Fastener links should be strong and reliable, and there should be no looseness.

Instrument Function Test after Experiment

1）For sample processing before pathological analysis, fully automated completion including dewaxing, antigen retrieval, staining, and fluorescence in situ hybridization (FISH ) pre-treatment, post-hybridization washing；

2）The fully automated immunohistochemical staining machine has36staining positions；

3）The fully automated immunohistochemical staining machine has40reagent positions；

4）Equippedwith a slide QR codescanning system；

5）The software interface can display the buffer volume in real time；

6）It has the function of classifying and storing waste liquid；

7）Has a patient information management system；

8）Has reagent information management system

Instrument performance testing after the trial:

1）The temperature control range of the dyeing module heating component is:37℃、65℃、85℃、101℃。

Temperature accuracy is ±2℃, temperature fluctuation is less than 0.5℃.

2）Reagent addition amount

The reagent addition volume control range is:100μL、130μL、240μL、370μL、500μL。

Accuracy of addition≤3%, coefficient of variation CV≤5%.

Chapter 3 Verification Records

3.1 Verification results

3.2 Test data records

3.2.1TestVibration, Collision,Simulated Transportand DropTest Before/AfterPerformance Test Data

Table3.2.1.1 Pre-testTemperature Accuracy

Table3.2.1.2 Temperature FluctuationBefore Testing

Table3.2.1.3 Temperature AccuracyAfterTest

Table3.2.1.4 Post-TestTemperatureFluctuation