M494E Biostation IM Perfusion Unit

7/23/2019 M494E Biostation IM Perfusion Unit

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Time Lapse Imaging System

BioStation IM

BS-IM Perfusion Unit

Instructions

M 9 E 08.6.NF.1

*M494EN01*


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i

Introduction

Thank you for purchasing the Nikon products.

This instruction manual is written for the users of the BS-IM Dispense Unit, an accessory for the Time lapse imaging

system BioStation IM.

To ensure correct usage, read this manual carefully before operating the product.

No part of this manual may be reproduced or transmitted in any form without prior written permission from Nikon.

The contents of this manual are subject to change without notice.

Although every effort has been made to ensure the accuracy of this manual, errors or inconsistencies may remain. If

you note any points that are unclear or incorrect, please contact your nearest Nikon representative.

Some of the equipment described in this manual may not be included in the set you have purchased.

If you intend to use any other equipment with this product, read the manual for that equipment too.

If the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may

be impaired.

Microsoft, Windows, and Internet Explorer are registered trademarks of Microsoft Corporation in the U.S. and other

countries. Other product and company names mentioned in this manual are trademarks or registered trademarks of

their respective owners.


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ii

Safety Precautions

To ensure correct and safe operation, read this manual before using the product.

Warning and Caution Symbols Used in This Manual

Although this product is designed to be completely safe during use, incorrect usage or failure to follow the safety

instructions provided may cause personal injury or property damage. To ensure correct usage, read this manual carefully

before using the product. Do not discard this manual and keep it handy for easy reference.

Safety instructions in this manual are marked with the following symbols to highlight their importance. For your safety,

always follow the instructions marked with these symbols.

Symbol Description

WARNINGDisregarding instructions accompanying this symbol may lead to serious injury or death.

CAUTIONDisregarding instructions accompanying this symbol may lead to injury or property

damage.


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Safety Precautions

iii

WARNING

1. Intended application of this product

The BS-IM Dispense Unit is a unit attached inside the BioStation IM (temperature controller area),

allowing the user to inject or to replace test fluids and fluid substances without opening/closing the door

or culture chamber. Do not use this product for other purpose.

The product is classified as an in-vitro diagnostic device.

2. Do not disassemble.

Disassembling may cause malfunction and will lead to the forfeiture of all claims against warranty. Do not

disassemble any part that is not indicated in this manual. If you notice any abnormality, contact your

nearest Nikon representative.

3. Read the instructions carefully.To ensure safety, carefully read this manual and the manuals for other equipment used with this product.

Especially be sure to follow the warnings and cautions indicated in the first of the manual.

4. Note on handing a hazardous specimen

Before handling a biological specimen, its risk must be checked.

When a hazardous specimen is used with this product, consult your safety supervisor or safety standard

of your facility. When a potentially infectious specimen is used, you must wear rubber gloves to avoid

direct touch.

If such specimen is spilled onto this product, the portion must be decontaminated in a safety manner.

Consult your safety supervisor or safety standard of your facility.

CAUTION

1. Disposal of the product

To avoid biohazard risks, dispose of the product as contaminated equipment according to the standard

procedure specified for your facility.


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Safety Precautions

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Notes on Handling the Product

1. Handling the Product.

This product is a precision instrument. Handle the product with care to avoid a physical shock or impact.

Cautions on moving the product

Remove the test fluid bottle and disconnect the tubes before carrying the BioStation IM with the Dispense Unit

attached.

Cautions on assembling and installing the product

Take care to avoid pinching your finger or hand.

Scratches or fouling such as fingerprints on optical components (such as lenses or filter) will degrade

microscope images.

Be careful to avoid scratches or direct contact with the objective when assembling the Dispense Unit.

2. Installation location and storage location

This product is an accessory of the BioStation IM. Select an installation location and a storage location as specified

for the BioStation IM. For details, refer to Safety Precautions in the separate manual BioStation IM Instructions

.


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v

ContentsIntroduction ............................................................................................................................................................ i

Safety Precautions ................................................................................................................................................ ii

Chapter 1 Equipment Overview......................................................................................................................... 1

1.1 Components .............................................. .................................................... ................................ 1

1.1.1

Perfusion induction unit (MFC99010) ................................................ ................................. 1

1.1.2

Perfusion medium exchange unit (MFC99015) ..................................................... ............. 2

1.2 Overview of the Perfusion Induction Unit................................... .................................................... 3

1.3 Overview of the Perfusion Medium Exchange Unit............ ........................................................ .... 4

1.3.1 Medium exchange - [degraded culture mediumfresh culture medium].......................... 5

1.3.2 Medium exchange - [culture mediumculture medium containing test fluid] ................... 6

1.3.3 Medium exchange

- [culture mediumculture medium containing test fluid culture medium] ...................7

Chapter 2 Names and Functions of Each Part ................................................................................................. 8

2.1

Parts Names and Functions of the Perfusion Induction Unit............................................. ............. 8

2.2 Parts Names and Functions of the Perfusion Medium Exchange Unit - [degraded culture medium

fresh culture medium] and [culture medium culture medium containing test fluid]................ 9

2.3 Parts Names and Functions of the Perfusion Medium Exchange Unit

- [culture mediumculture medium containing test fluidculture medium] ............................ 10

Chapter 3 Attaching to the BioStation IM ....................................................................................................... 11

3.1 Attaching the Perfusion Induction Unit.......................................................... ............................... 11

3.1.1 Removing the culture chamber from the BioStation IM .................................................... 11

3.1.2 Attaching the dispense needle assembly and bottle holder to the culture chamber ......... 13

3.1.3 Attaching the culture chamber to the BioStation IM..................................................... ..... 14

3.1.4

Installing the bottle and connecting the tubes and fittings ................................................ 15

3.2 Attaching the Perfusion Medium Exchange Unit

(for [degraded culture mediumfresh culture medium] / [culture mediumculture medium

containing test fluid] setup) ................................................... ....................................................... 17

3.2.1 Removing the culture chamber from the BioStation IM .................................................... 17

3.2.2 Attaching the dispense/drain needle assembly and connecting tubes ............................. 19

3.2.3

Attaching the culture chamber to the BioStation IM..................................................... ..... 22

3.2.4 Connecting tubes from outside of the BioStation IM.................................................... ..... 23

3.2.5 Piping arrangement for medium exchange

(for [degraded culture mediumfresh culture medium] /

[culture mediumculture medium containing test fluid] setup) ...................................... 25

3.3 Adjusting the Needle Position......................... ....................................................... ...................... 26

Attaching the Perfusion Medium Exchange Unit ......................................................... ............... 27

(for [culture mediumculture medium containing test fluid culture medium] setup)................. 27

1.

Removing the culture chamber from the BioStation IM .................................................... 27

2. Attaching the dispense/drain needle assembly and connecting tubes ............................. 27

3. Attaching the culture chamber to the BioStation IM..................................................... ..... 32

4. Connecting tubes from outside of the BioStation IM.................................................... ..... 32

5. Piping arrangement for medium exchange

(for [culture medium

culture medium containing test fluid

culture medium] setup)......... 34

Chapter 4 Operating Procedures..................................................................................................................... 35

4.1 Filling the Bottle with Culture Medium................................................ .......................................... 35


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Contents

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4.2 Setting and Removing the Specimen........................................... ................................................ 37

4.2.1

Setting the specimen.......... ....................................................... ....................................... 37

4.2.2 Removing the Specimen ................................................... ............................................... 38

4.3 Test Fluid Dispense/Medium Exchange ................................................ ...................................... 39

4.3.1

Dispensing procedure for test fluid, etc. ........................................................ ................... 39

4.3.2 Draining procedure for culture medium, etc...................................................................... 40

4.3.3 Focus adjustment ................................................... ....................................................... ... 40

4.3.4 Use of peristaltic pumps ................................................... ................................................ 40

Chapter 5 Care and Maintenance .................................................................................................................... 41

5.1 Cleaning ............................................ ..................................................... ..................................... 41

5.2 Parts Treatable in Autoclave Sterilization ................................................... ................................. 41

Chapter 6 Specifications.................................................................................................................................. 42


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1

Equipment Overview11

The BS-IM Perfusion Unit is a unit attached inside the BioStation IM (temperature controller area), allowing the user to

inject or to replace test fluids and fluid substances without opening/closing the door or culture chamber.

All the components of the unit are attached to the interior of the temperature controlled area of the BioStation IM. This

configuration eliminates the need for an additional device to warm replacement culture medium, and maintains the

environment of tubes connected to the dish at a controlled temperature.

1.1 Components

This option unit includes perfusion induction unit and a perfusion medium exchange unit.

1.1.1 Perfusion induction unit (MFC99010)

This is the basic unit for the BS-IM Perfusion Unit. Mounting the perfusion induction unit to the BioStation IM culture

chamber allows injection of test fluid to the culture medium without taking the dish out of the culture chamber.

Components in the perfusion induction unit

Dispense needle assembly(BS-P1): x1 Bottle: x1

Tube Clamp (blue): x1 Fitting (female): x1

Silicon tube (1100 mm): x1 Fitting (male) + silicon tube: x1

L-shaped fitting + silicon tube: x2


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Chapter 1 Equipment Overview

2

1.1.2 Perfusion medium exchange unit (MFC99015)

This unit, either partly or entirely, is used in combination with the perfusion induction unit. The perfusion medium

exchange unit alone does not provide any function. The use of this unit allows culture medium replacement without

taking the dish out of the culture chamber.

Components in the perfusion medium exchange unit

Drain needle assembly

(BS-P2): x1

Bottle: x1

Tube clamp

(red, yellow): x2

Fitting (female): x2

Silicon tube (1100 mm): x2

Fitting (male): x1

T-shaped fitting

+ L-shaped fitting

+ silicon tube: x1

Silicon tube (1100 mm): x2

Bottle holder: x1 Check valve: x2 Clamp: x2

Hexagonal socket

head bolt (M3 x 4): x2

L-shaped fitting+ silicon tube: x2


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1.2 Overview of the Perfusion Induction Unit

This arrangement allows only test fluid injection to the culture medium.

Operators can observe the sample reactions induced by test fluid injection.

Figure 1.1-1 Overview of the perfusion induction unit

Test fluid bottleCulture chamber

Dish

Objective

Dispense

needle

BioStation IM (slide door)

Pressure applied

(from a syringe or peristaltic

pump) to send test fluid


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1.3 Overview of the Perfusion Medium Exchange Unit

This unit allows users to exchange culture medium without taking the dish out of the culture chamber.

This unit and the perfusion induction unit can be configured in two ways, and the two configurations can be used for

three different applications.

Table 1.1-1 Medium exchange combinations

PartQua

ntity

Medium exchange -

[degraded culture medium

fresh culture medium] and

[culture mediumculture

medium containing test fluid]

Medium exchange


medium containing test

fluidculture medium]

Dispense needle assembly x1

Bottle x1

Tube clamp (blue) x1

Fitting (female) x1

L-shaped fitting + silicon tube x2 X

Fitting (male) + silicon tube x1

Perfus

ioninductionunit

Silicon tube (1100 mm) x1

Drain needle assembly x1

Bottle x1 X

Bottle holder x1 X

Tube clamp (red, yellow) x2 (x1)

Fitting (female) x2 (x1)

Fitting (male) x1

Silicon tube (1100 mm) x2 (x1)

Silicon tube (110mm) x2

Hexagonal socket head bolt x2

Check valve x2

Clamp (for check valve) x2

L-shaped fitting + silicon tube x2

Per

fusionmediumexchangeunit

T-shaped fitting + L-shaped fitting+ silicon tube

x1

= used / X = not used / = partially used


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Table 1.1-2 Medium exchange applications

Name Application

Medium exchange -

[degraded culture medium

fresh culture medium]

Drain the degraded culture medium launch fresh culture medium

Drain the culture medium degraded over prolonged culturing from the dish, and

launch fresh culture medium.

Medium exchange -



Drain culture medium launch culture medium containing test fluid

Drain the culture medium in the dish, and launch culture medium containing test

fluid.

Medium exchange -


medium containing test fluid

culture medium]

Drain culture medium launch culture medium containing test fluid drain

culture medium containing test fluid launch culture medium.

Drain the culture medium in the dish, and launch culture medium containing test

fluid. Drain the culture medium containing test fluid in the dish, and launch

culture medium.

1.3.1 Medium exchange - [degraded culture mediumfresh culture medium]

After draining degraded culture medium in the dish, launch fresh culture medium.

Culture medium is exchanged by draining the dish of culture medium degraded over prolonged culturing, and then

injecting fresh culture medium. This enables culturing for an extended period.

Figure 1.1-2 Medium exchange - [degraded culture mediumfresh culture medium]

Culture bottle

Culture chamber

Dish

Objective

Dispense

needle


Pressure applied (from a

syringe or peristaltic pump)

to send fresh culture medium



to drain degraded culture

medium

Drain needle


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1.3.2 Medium exchange - [culture mediumculture medium containing test fluid]

After draining the culture medium in the dish, launch culture medium containing test fluid.

This is mechanically identical to the medium exchange - [degraded culture medium fresh medium]. Drain the culture

medium in the dish, then launch culture medium containing test fluid. This system allows the user to steadily manage theconcentration of the test substance and the time required to reach the sample.

Figure 1.1-3 Medium exchange - [culture mediumculture medium with test fluid]

Bottle of culture

medium with test fluid

Culture chamber

Dish

Objective

Dispense

needle



syringe or peristaltic pump)to send culture medium

containing test fluid


syringe or peristaltic pump)to drain culture medium

Drain needle


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1.3.3 Medium exchange -

[culture mediumculture medium containing test fluid culture medium]

After draining the culture medium, launch the culture medium containing a desired test fluid. Drain the culture medium

containing the test fluid to launch the culture medium.Two bottles are provided with this exchange unit: One is applied for the culture medium, the other one for the culture

medium containing the test fluid. After draining the culture medium from the dish, inject the culture medium containing

the test fluid. After observing sample reaction to the test substance, repeat the procedures of draining and injecting the

culture medium until almost any test substance is remained in the dish. In this way, you can verify if the test results are

accurate.

Figure 1.1-4 Medium exchange - [culture mediumculture medium containing test fluid culture medium]

Bottle of culturemedium containingtest fluid

Culture chamber

Dish

Objective

Dispense

needle




to send culture medium



syringe or peristaltic

pump) to drain culture

medium/ culture medium


Drain needle

Culture bottle

Check valve

Pressure applied

(from a syringe or

peristaltic pump)

to send culture

medium

Check valve


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Names and Functions of Each Part22

This chapter explains the name and function of each part in both units.

2.1 Parts Names and Functions of the Perfusion Induction Unit

This setup is for test fluid dispense only. The unit consists only of the parts used for test fluid dispense.

Figure 2.1-1 Unit configuration

Table 2.1-1 Parts names and functions of the perfusion induction unit

Name Function

Dispense needleassembly

Needle assembly for dispensing test fluid into the dish.The position of the dispense needle can be adjusted axially and vertically. Additionally, the

dispense needle can be set in the dispense position or the standby position. The needle

should be set in the standby position for changing dishes.

Piping parts Silicon tube and couplings.

Bottle 1 bottle for holding the test fluid is installed.

*Culture chamber and dish are not included in this product.

Bottle

Silicon tube (1100 mm)

Fitting (male) + silicon tube

Dispense needle assembly

L-shaped fitting + silicon tube

Tube clamp (blue)

Fitting (female)


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Chapter 2 Names and Functions of Each Part

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2.2 Parts Names and Functions of the Perfusion Medium Exchange Unit -[degraded culture mediumfresh culture medium] and [culture mediumculture medium containing test fluid]

This setup is for [draining degraded culture medium and injecting fresh culture medium], or [draining culture medium and

injecting culture medium containing a test fluid].

Components of both the perfusion induction unit and the exchange unit are used for this configuration.

Figure 2.1-2 Unit configuration [degraded culture mediumfresh culture medium] and [culture medium culture


Table 2.1-2 Parts names and functions of the perfusion medium exchange unit [degraded culture mediumfresh culture

medium] and [culture mediumculture medium containing test fluid]

Name Function

Dispense needle

assembly

Needle assembly for dispensing culture medium or culture medium containing test fluid into the dish.

The position of the dispense needle can be adjusted axially and vertically. Additionally, the

dispense needle can be set in the dispense position or the standby position. The needle should

be set in the standby position for changing dishes.

Drain needle

assembly

Needle assembly for draining culture medium or degraded culture medium from the dish.

The position of the drain needle can be adjusted axially and vertically. Additionally, the drain

needle can be set in the drain position or the standby position. Both the dispense and drain

needles should be set in the standby position for changing dishes.


Bottle 1 bottle for holding the culture medium or culture medium containing test fluid is installed.


Bottle





Tube clamp (blue)

Fitting (female)

Rotatable cover stopper

Fitting (female)Drain needle assembly


Fitting (male) Tube clamp (red or yellow)


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Chapter 2 Names and Functions of Each Part

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2.3 Parts Names and Functions of the Perfusion Medium Exchange Unit - [culturemediumculture medium containing test fluidculture medium]

This setup is for [draining culture mediumdispensing culture medium containing test fluid draining culture medium


dispensing culture medium].All the components for both the perfusion induction unit and the exchange unit are used for this configuration.

Figure 2.1-3 Unit configuration [culture mediumculture medium containing test fluidculture medium]

Table 2.1-3 Parts names and functions of the perfusion medium exchange unit [culture mediumculture medium containing

test fluidculture medium]

Name Function

Dispense needle

assembly

Needle assembly for dispensing culture medium or culture medium containing test fluid into the

dish.

The position of the dispense needle can be adjusted axially and vertically. Additionally, the

dispense needle can be set in the dispense position or the standby position. Both the dispense

and drain needles should be set in the standby position for changing dishes.

Drain needle

assembly

Needle assembly for draining culture medium or degraded culture medium from the dish.

The position of the drain needle can be adjusted axially and vertically. Additionally, the drain

needle can be set in the drain position or the standby position.

Both the dispense and drain needles should be set in the standby position for changing dishes.

Check valve Prevents the culture medium and culture medium containing test fluid from mixing.


Bottle Two bottles for holding the culture medium and culture medium containing test fluid are installed.


Bottle




L-shaped fitting +

silicon tube

Tube clamp (blue)

Fitting (female)

Tube clamp (red or yellow)

Fitting (female)

Drain needle assembly


Fitting (male)



Check valve

Clamp

T-shaped fitting +

L-shaped fitting +

silicon tube

Bottle holder


Rotatable cover stopper


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Attaching to the BioStation IM33

3.1 Attaching the Perfusion Induction Unit

Necessary tool

The following tool is required for the procedure below.

Hexagonal screwdriver (supplied with BioStation IM)

3.1.1 Removing the culture chamber from the BioStation IM

When installing the Perfusion Unit to the culture chamber, remove the culture chamber from the BioStation IM in

advance.

1. Open the slide door on the BioStation IM.

Figure 3.1-1 Opening the slide door

2. Move the diascopic illuminator and duct out of

the way.

Figure 3.1-2 Moving the diascopic illuminator and

duct out of the way

3. Remove the temperature sensor.

Figure 3.1-3 Removing the temperature sensor

Slide door

Diascopic illuminator

Duct

Temperature sensor


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Chapter 3 Attaching to the BioStation IM

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4. Remove the exhaust tube and wet mixed gas

tube.

Figure 3.1-4 Removing the tubes

5. Remove the culture chamber.

Remove the three (x3) hexagonal socket head bolts using

the hexagonal screwdriver.

Figure 3.1-5 Removing the culture chamber

Exhaust tube

Hexagonal

socket head

bolt

Hexagonal socket head bolt

Wet mixed gas tube


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3.1.2 Attaching the dispense needle assembly and bottle holder to the culture chamber

Attach the dispense needle assembly and bottle holder to the culture chamber that was removed earlier.

1. Remove the right side plate of the culturechamber.

Remove the two (x2) hexagonal socket head bolts using


Be careful not to lose the removed plate or hexagonal

socket head bolts.

Figure 3.1-6 Removing the right side plate of

culture chamber

2. Attach the dispense needle assembly to the

culture chamber.

Attach the dispense needle assembly to the culture

chamber using the two (x2) hexagonal socket head bolts.

Figure 3.1-7 Attaching the dispense needle

assembly

CAUTION

Caution in passing the dispense needle through the opening in the culture chamber

When passing the dispense needle through the culture chamber, be careful not to bump the needle tip against the

culture chamber. This assembly may fail to operate properly, if the dispense needle is deformed.

Hexagonal socket

head bolt

Hexagonal socket

head bolt

Plate

Hexagonal

socket head

bolt


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3.1.3 Attaching the culture chamber to the BioStation IM

Attach the culture chamber preassembled with the dispense needle assembly to the BioStation IM.

1. Attach the culture chamber.Attach the culture chamber to the BioStation IM using the

three (x3) hexagonal socket head bolts.

Figure 3.1-8 Attaching the culture chamber

2. Connect the exhaust tube and wet mixed gas

tube.

Figure 3.1-9 Connecting the tubes

3. Attach the temperature sensor.

Figure 3.1-10 Attaching the temperature sensor

Hexagonal socket

head bolt

Exhaust tube

Temperature sensor


Wet mixed

gas tube


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3.1.4 Installing the bottle and connecting the tubes and fittings

Place the bottle into the bottle holder and connect the tubes and fittings.

1. Place the bottle into the bottle holder.

Figure 3.1-11 Installing the bottle

2. Connect the tubes and fittings.

Connect the tubes and fittings as shown in the figure

below.

Figure 3.1-12 Connecting the tubes and fittings

Bottle




Fitting (female)

Tube clamp (blue)


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CAUTION

Caution in connecting the tubes and fittings

Make sure all tubes and fitting couplings are tightened. Test fluid or culture medium may leak from loose tube

connections and cause contamination or damage this instrument.

Pass the silicon tube (1100 mm x1) for dispense through

the points shown in the figure on the right and pull the tube

out of the BioStation IM.

Figure 3.1-13 Routing silicon tubes

CAUTION

Caution in pulling out the silicon tube

When pulling the silicon tube (1100 mm) out of the BioStation IM, be careful not to pinch it in the front door of the

BioStation IM.

Silicon tube

(1100 mm)

for dispense


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3.2 Attaching the Perfusion Medium Exchange Unit(for [degraded culture mediumfresh culture medium] / [culture mediumculture medium containing test fluid] setup)

This section describes the installation procedure for the perfusion medium exchange setup for [degraded culture medium

fresh culture medium] and [culture medium culture medium containing test fluid].

For installation procedures of the perfusion medium exchange setup for [culture mediumculture medium containing

test fluidculture medium], refer to Section 3.4, Attaching the Perfusion Medium Exchange Unit (for [culture medium

culture medium containing test fluidculture medium]).

Necessary tool

The following tool is required for the procedure below.

Hexagonal screwdriver (supplied with BioStation IM)

3.2.1 Removing the culture chamber from the BioStation IM

When installing the Perfusion Unit to the culture chamber, remove the culture chamber from the BioStation IM in

advance.


Figure 3.2-1 Opening the slide door

2. Move the diascopic illuminator and duct out of

the way.

Figure 3.2-2 Moving the diascopic illuminator and

duct out of the way

Slide door

Diascopic illuminator

Duct


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3. Remove the temperature sensor.

Figure 3.2-3 Removing the temperature sensor

4. Remove the exhaust tube and wet mixed gas

tube.

Figure 3.2-4 Removing the tubes

5. Remove the culture chamber.Remove the three (x3) hexagonal socket head bolts using


Figure 3.2-5 Removing the culture chamber

Temperature sensor

Exhaust tube

Hexagonal

socket head

bolt


Wet mixed gas tube


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3.2.2 Attaching the dispense/drain needle assembly and connecting tubes

Attach the dispense needle assembly and drain needle assembly to the culture chamber that was removed earlier.

1. Remove the right and left side plates of theculture chamber.

Remove the two (x2) hexagonal socket head bolts on each

side using the hexagonal screwdriver.


socket head bolts.

Of the plates removed, the left plate is made taller than the

right plate to serve as a stopper for the rotatable cover.

Make sure to return the plates to their correct positions

when reassembling.Figure 3.2-6 Removing the right and left side plates

of culture chamber


culture chamber.



Figure 3.2-7 Attaching the dispense needle

assembly

CAUTION





Plate

Hexagonal socket

head bolt

Hexagonal

socket headbolt



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3. Attach the drain needle assembly to the

culture chamber.

Attach the drain needle assembly to the culture chamber

using the two (x2) special hexagonal socket head bolts.

Figure 3.2-8 Attaching the drain needle assembly

CAUTION

Caution in passing the drain needle through the opening in the culture chamber

When passing the drain needle through the culture chamber, be careful not to bump the needle tip against the

culture chamber. This assembly may fail to operate properly, if the drain needle is deformed.

4. Adjust the height of the drain needle.

The height and axial position of both needles are adjustable. Normally, to attach the drain needle, adjust the height

of the drain needle. Apply additional adjustments if necessary.For details on height adjustment procedures for the drain needle, refer to Section 3.3 "Adjusting the Needle

Position".

5. Connect the tubes and fittings to the bottle.

1) Connect the L-shaped fittings and silicon tubes (2 sets)

to the bottle.

2) Connect the male fitting and silicon tube to the

L-shaped fitting and silicon tube on the dispensing side

of the bottle (with a tube inside).

Figure 3.2-9 Connecting tubes to the bottle

CAUTION


Make sure all tubes and fittings couplings are tightened. Test fluid or culture medium may leak from loose tubeconnections and cause contamination or damage this instrument.

Hexagonal socket

head bolt

Hexagonalsocket head bolt

Fitting (male)

+ silicon tube

L-shaped fitting

+ silicon tube


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6. Place the bottle into the bottle holder and

connect the tubes.

Place the bottle with tubes connected into the bottle holder

and insert the male fitting into the dispense needle.

Figure 3.2-10 Installing and connecting tubes to the

bottle

Fitting (male)

+ silicon tube

Bottle


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3.2.3 Attaching the culture chamber to the BioStation IM

Attach the culture chamber preassembled with the dispense needle assembly and drain needle assembly to the

BioStation IM.

Figures below illustrate setup for medium exchange - [culture mediumculture medium containing test fluid]. Setupprocedure for this application is the same as the procedure for medium exchange - [culture mediumculture medium

containing test fluidculture medium].

1. Attach the culture chamber.

Attach the culture chamber to the BioStation IM using the

three (x3) hexagonal socket head bolts.

Figure 3.2-11 Attaching the culture chamber

2. Connect the exhaust tube and wet mixed gas

tube.

Figure 3.2-12 Connecting the tubes

Hexagonal socket

head bolt

Exhaust tube

Hexagonal socket

head bolt

Wet mixed

gas tube


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3. Attach the temperature sensor.

Figure 3.2-13 Attaching the temperature sensor

3.2.4 Connecting tubes from outside of the BioStation IM

Connect the silicon tubes for dispense and drain from outside of the BioStation IM.

1. Insert the silicon tubes for dispense and drain.

Pass the silicon tubes (1100 mm) for dispense (x1) and

drain (x1) through the points shown in the figure on the

right and pull the tubes into the BioStation IM.

Figure 3.2-14 Routing silicon tubes

CAUTION



BioStation IM.

Temperature

sensor

Silicon tube

(1100 mm)

for dispense

Silicon tube

(1100 mm)

for drain


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2. Connect each silicon tube (1100 mm) to the BS-IM Dispense Unit.

Test fluid dispense tube

Connect the silicon tube (1100 mm) to the L-shaped fitting and silicon tube on the air pressure side of the bottle

(with no tube inside).

Drain tube

Connect the male fitting to the tip of the silicon tube (1100 mm) and connect the male fitting to the drain needle.

Figure 3.2-15 Connecting the silicon tubes for dispense and drain

CAUTION


Make sure all tubes and fittings couplings are tightened. Test fluid or culture medium may leak from loose tube


3. Attach the male fitting and tube clamp to the

silicon tubes for dispense and drain.

Three tube clamps are provided: blue, red and yellow. Besure that each silicon tube is recognized when attaching it.

Figure 3.2-16 Attaching the fitting and the tube

clamp


for dispense

Silicon tube (1100 mm) for drain

Fitting (male)

Tube clamp

Fitting (female)


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3.2.5 Piping arrangement for medium exchange

(for [degraded culture mediumfresh culture medium] / [culture medium culture

medium containing test fluid] setup)

The piping arrangement for medium exchange [degraded culture mediumfresh culture medium] and [culture mediumculture medium containing test fluid] is as shown below.

After connecting the tubes, make sure that the parts and tubes are connected correctly.

Figure 3.2-17 Connecting the tubes and fittings

CAUTION

Caution in connecting tubes to the bottle

Be careful not to connect the wrong tube to the bottle.The inlet with a tube: connect the silicon tube that leads to the dispense needle

The inlet without a tube inside: connect the silicon tube (1100 mm) that leads to outside the BioStation IM.




Fitting (male)

Silicon tube (1100mm)

Fitting (female)

Fitting (female)

Tube clamp (blue)



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3.3 Adjusting the Needle Position

The needle has adjustments for height and axial positioning.

Normally, only the height of the drain needle needs to be adjusted. The axial position of the drain needle and the

dispense needle position should not require adjustment. Make these adjustments if needed.

Example below describes how to adjust the position for the drain needle, which procedure is also applied to adjust the

position for the dispense needle.


1) Slightly loosen the hexagonal socket head bolts that

hold the drain needle assembly in place on the left and

right sides.

2) To adjust the clearance between the bottom surface of

the dish and the tip of the drain needle, gently shake

the left side of the mounting piece for the culture

chamber up and down using the hexagonal socket

head bolt on the right side as the pivot.

3) Once adjusted, fix the culture chamber in place by

tightening the left and right hexagonal socket head

bolts.

Figure 3.3-1 Adjusting the height of drain needle

Adjusting the height of the drain needle

The drain needle is made slightly longer than the dispense needle.

When adjusting the height of the drain needle, aim to put the needle tip as close to the bottom of the dish aspossible. If the drain needle tip is off the bottom of the dish, the more the amount of culture medium or test fluid left

undrained in the dish.

2. Adjust the axial position of the drain needle.

1) Loosen the hexagonal socket head bolt (x1) in the top

of the needle assembly using the hexagonal

screwdriver.

2) Slide the cover on the needle in the axial direction to

adjust the position of the tip of the drain needle.

3) Once adjusted, tighten the hexagonal socket head bolt

(x1) in the top of the needle assembly using the

hexagonal screwdriver.Figure 3.3-2 Adjusting the axial position of drain

needle

Hexagonal socket head bolt (pivot side)

Hexagonal socket

head bolt

Hexagonal socket head bolt (adjustment side)

Cover on

needle

assembly


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Attaching the Perfusion Medium Exchange Unit(for [culture mediumculture medium containing test fluidculture medium] setup)

1. Removing the culture chamber from the BioStation IM

When installing the Perfusion Unit to the culture chamber, remove the culture chamber from the BioStation IM in advance.

Refer to Section 3.2.1, Removing the culture chamber from the BioStation IM for details of the procedure.

2. Attaching the dispense/drain needle assembly and connecting tubes

Attach the dispense needle assembly, drain needle assembly, and bottle holder to the culture chamber that was removed

earlier.

1. Remove the bottle holder from the dispense

needle assembly.Remove the hexagonal socket head bolt using the

hexagonal screwdriver.

Figure 2-1 Removing the bottle holder

2. Combine the bottle holders.

Take the bottle holder removed from the dispense needle

assembly and combine it with the bottle holder included in

the perfusion medium exchange unit using the two (x2)

hexagonal socket head bolts.

Figure 2-2 Combining the bottle holders

3. Attach the clamp for the check valve to the

combined bottle holders.

Secure the clamp by inserting the tab on the clamp into

the slotted mounting hole. With the clamp pressed against

the bottle holder, pull upwards.

Figure 2-3 Attaching the clamps


Hexagonal sockethead bolt

Clamp

Bottle holder

Hexagonal socket

head bolt

Clamp


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4. Connect the tubes and fittings to the check

valve.

When connecting, the arrows on the check valves should

be pointed at the T-shaped fitting, L-shaped fitting, andsilicon tube.

Figure 2-4 Connecting the tubes and fittings to

the check valve

CAUTION

Caution in connecting the tubes and fittings to the check valve

Be sure that the arrow on the check valve faces the correct direction. The check valve will not function if it is

connected in the reverse direction.

5. Connect the check valve.

Attach the check valves to each clamp as shown in the

figure on the right, and then pass the tube through the

bottle holders.

Figure 2-5 Connecting the check valves

CAUTION

Caution in connecting the check valves to the clamps

Attach each check valve so that the ribs touch the inside of the clamp. The check valve is not fixed in place

because the recessed surfaces do not touch the inside of the clamp.

6. Attach the assembled bottle holders to the

dispense needle assembly.

Attach the assembled bottle holders to the dispense

needle assembly using the hexagonal socket head bolt

(x1).

Figure 2-6 Attaching the bottle holders

T-shaped fitting + L-shaped fitting + silicon tube


Check valve

Check valve

Dispense needle

assembly


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7. Remove the right and left side plates of the

culture chamber.

Remove the two (x2) hexagonal socket head bolts on each

side using the hexagonal screwdriver.


socket head bolts.

Of the plates removed, the left plate is made taller than the

right plate to serve as a stopper for the rotatable cover.

Make sure to return the plates to their correct locations

when reassembling.Figure 2-7 Removing the right and left side plates

of culture chamber


culture chamber.



Figure 2-8 Attaching the dispense needle

assembly

CAUTION




Hexagonal socket

head bolt

Hexagonal socket

head bolt


Plate

Hexagonal

socket head

bolt


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9. Attach the drain needle assembly to the

culture chamber.

Attach the drain needle assembly to the culture chamber

using the two (x2) special hexagonal socket head bolts.

Figure 2-9 Attaching the drain needle assembly

CAUTION

Caution in passing the drain needle through the opening in the culture chamber

When passing the drain needle through the culture chamber, be careful not to bump the needle tip against the

culture chamber. This assembly may fail to operate properly, if the drain needle is deformed.


The height and axial position of both needles are adjustable. Normally, to attach the drain needle, adjust the heightof the drain needle. Apply additional adjustments if necessary.

For details on height adjustment procedures for the drain needle, refer to Section 3.3 Adjusting the Needle

Position.

11. Connect the tube for the dispense needle.

Connect the T-shaped tube assembly to the dispense

needle with the male fitting and silicon tube.

Figure 2-10 Tube connections for the dispense

needle

Hexagonal socket

head bolt

Hexagonal

socket

head bolt

Fitting (male)

+ silicon tube


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12. Connect the tubes and fittings to each bottle.

Connect the L-shaped fitting and silicon tube to the air

pressure inlet (without a tube) of each bottle.

Figure 2-11 Connecting tubes and fittings to the

bottle

13. Install the bottles.

Place each bottle in the bottle holder.

Figure 2-12 Placing the bottle

14. Connect the tube between the check valve and

bottle.

Connect the silicon tube (110 mm) between the inlet fitting

(with the tube inside) of the test fluid dispense bottle and

the check valve.

Figure 2-13 Placing the bottle and connecting

tubes

L-shaped fitting

+ silicon tube

BottleBottle

Silicon tube

(110 mm)


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3. Attaching the culture chamber to the BioStation IM

Attach the culture chamber preassembled with the dispense needle assembly and drain needle assembly to the

BioStation IM.

Refer to Section 3.2.3, Attaching the culture chamber to the BioStation IM for details of the procedure.

4. Connecting tubes from outside of the BioStation IM

Connect the silicon tubes for dispense and drain from outside of the BioStation IM.

1. Insert the silicon tubes for dispense and drain.

Pass the silicon tubes (1100 mm) for dispense (x2) and

drain (x1) through the points shown in the figure on the

right and pull the tubes into the BioStation IM.

Figure 4-1 Routing silicon tubes

CAUTION



BioStation IM.

Silicon tube

(1100 mm)

for dispense

Silicon tube

(1100 mm)

for drain


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2. Connect each silicon tube (1100 mm) to the BS-IM Dispense Unit.

Test fluid dispense tube

Connect the silicon tube (1100 mm) to the L-shaped fitting and silicon tube on the air pressure side of the bottle

(with no tube inside).

Drain tube

Connect the male fitting to the tip of the silicon tube (1100 mm) and connect the male fitting to the drain needle.

Figure 4-2 Connecting the silicon tubes for dispense and drain

CAUTION


Make sure all tubes and fittings couplings are tightened. Test fluid or culture medium may leak from loose tube


3. Attach the male fitting and tube clamp to the

silicon tubes for dispense and drain.

Three tube clamps are provided: blue, red and yellow. Besure that each silicon tube is recognized when attaching it.

Figure 4-3 Attaching the fitting and the tube clamp

Silicon tube (1100 mm) for dispense

Silicon tube (1100 mm) for drain

Fitting (male)

Tube clamp

Fitting (female)


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5. Piping arrangement for medium exchange

(for [culture mediumculture medium containing test fluid culture medium] setup)

The piping arrangement for medium exchange [culture mediumculture medium containing test fluid culture

medium] is as shown below.After connecting the tubes, make sure that the parts and tubes are connected correctly.

Figure 5-1 Connecting the tubes and fittings

CAUTION

Caution in connecting tubes to the bottle

Be careful not to connect the wrong tube to the bottle.

The inlet with a tube: connect the silicon tube that leads to the check valve.

The inlet without a tube inside: connect the silicon tube (1100 mm) that leads to outside the BioStation IM.



Connects the bottle and check valve


Connects the dispense

needle and T-shaped fitting


L-shaped fitting

+ silicon tube

Fitting (male)

Silicon tube

(1100mm)Fitting (female)

Fitting (female)

Tube clamp (blue)



Check valve

T-shaped fitting

+ L-shaped fitting

+ silicon tube


Connects the bottle

and check valve


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Operating Procedures44

This product does not have any pumps to dispense test fluid or drain culture medium.

Normally, a customer-provided syringe or peristaltic pump is used to dispense test fluid or drain culture medium.

4.1 Filling the Bottle with Culture Medium


2. Remove the bottle to fill with culture medium.

Disconnect and remove the silicon tube from the bottle

and remove the bottle.

Figure 4.1-1 Removing the bottle

3. Remove the lid from the bottle and fill with culture medium.

When removing the lid from the bottle, check if the packing inside the lid is not damaged.Be sure there are no

scratches or cracks or other signs of deterioration.

CAUTION

Packing used in the bottle lid

The bottle lid disassembles into the parts shown in the

figure on the right. The packing inside the lid should be

replaced if any deterioration is found. Make sure it is fit

in the lid after each replacement.

To ensure the bottle is sealed air tight, make sure thepacking is installed properly.

Figure 4.1-2 Packing in the bottle lid

Packing

Bottle lid

Silicon tube


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4. Return the bottle to the bottle holder and

reconnect the tubes.

Figure 4.1-3 Installing the bottle

5. Close the slide door on the BioStation IM.

CAUTION

Caution on the temperature of the culture medium/test fluid in the bottle

If the temperature of the culture medium or culture medium containing test fluid filled in the bottle is below

37.0 C, allow the content to reach 37.0 C before starting your experiment by placing the bottle in the BioStation

IM and allowing it to warm up.


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4.2 Setting and Removing the Specimen

When setting and removing the specimen (dish), both the dispense needle and drain needle must be in their standby

positions.

4.2.1 Setting the specimen

1. Open the slide door on the BioStation IM and move the diascopic illuminator out of the way.

CAUTION

Caution in moving the needle

When moving the needles to the standby, dispense or drain positions, always hold and turn the needle by the

metal base. Do not hold the silicon tubes, as this may loosen or disconnect the tube and cause it to break.

2. Move the drain needle and dispense needle to

their standby positions.

1) Push the needle slightly toward the culture chamber

until it moves out of the groove.

2) Turn the needle left 90 degrees.

3) Pull the needle toward you.

The needle can be pulled out to the position shown in

the figure below. Place the needle within the guide

groove unless otherwise desired.

Figure 4.2-1

CAUTION

Caution in pulling the needle to the limit.

If you close the slide door on the BioStation with the

needle pulled all the way out of the guide groove, the

door will hit the needle and possibly break it.

Figure 4.2-2 Moving the needle to the standby

position

1) Pull the needle out of the groove.

2) Turn the needle left 90 degrees.

3) Pull the needle toward you.


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3. Open the rotatable cover and set the dish.

Set the dish near the center of the culture chamber.

Figure 4.2-3 Setting the dish

4. Move the drain needle and dispense needle to

their drain and dispense positions,respectively.

Return each needle to their original positions by following

the procedures for moving them to the standby position in

reverse.

Figure 4.2-4 Moving each needle to the

drain/dispense positions

5. Close the rotatable cover on the culture

chamber.

Figure 4.2-5 Closing the rotatable cover on the

culture chamber

6. Return the diascopic illuminator to its original position and close the BioStation IM slide

door.

4.2.2 Removing the Specimen

Remove the specimen (dish) by following the setting procedures.

Dish


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4.3 Test Fluid Dispense/Medium Exchange

4.3.1 Dispensing procedure for test fluid, etc.

1. Connect a silicon tube to a syringe, and the

bottle containing test fluid or culture medium

mixed with test fluid.

Figure 4.3-1 Connecting to a pump

2. Dispense the required amount of test fluid

from the syringe.

Open the tube clamp when injecting, then close it

afterwards.

Figure 4.3-2 Dispensing test fluid, etc.

3. Record the timing of test fluid dispense in the

software.

- Cell stimulus mark setting button :

For an explanation of each symbol, refer to the separate

manual, BioStation IM Instructions , Chapter 4 Time-Lapse Images in Process

Screen, Section 4.1.8 Time Line and Image Playback

Control.Figure 4.3-3 Recording test fluid dispense

Syringe

Syringe

Fitting (female)

Tube clamp


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4.3.2 Draining procedure for culture medium, etc.

1. Connect the silicon tube from the discharge

needle to a syringe.

Figure 4.3-4 Connecting to a pump

2. Draw culture medium into the syringe.

Open the tube clamp when draining, then close it.

Figure 4.3-5 Draining culture medium, etc.

3. Record the timing of draining the culture

medium in the software, if necessary.

- Cell stimulus mark setting button :

For an explanation of each symbol, refer to the separate

manual, BioStation IM Instructions , Chapter 4 Time-Lapse Images in Process

Screen, Section 4.1.8 Time Line and Image Playback

Control.Figure 4.3-6 Recording draining of culture medium

4.3.3 Focus adjustment

After draining culture medium or dispensing test fluid to the dish, focus of the microscope may require adjustment as

focus can be affected by changes in weight of dish content or even by the slightest changes in temperature. Therefore,

verify the focus adjustment after medium exchange or test fluid dispense during time lapse examination. If the image

goes out of focus, readjust the Z coordinate as described in Section 2.4, Observation Point and Focus Adjustment in

the separate manual, BioStation IM Instructions .

4.3.4 Use of peristaltic pumps

To inject the test substance or to drain the culture medium, use a peristaltic pump instead of the syringe, if desired.

Culture medium launching time can be shortened when using a peristaltic pump by increasing the diameter of the silicon

tube with a commercially available conversion adapter.

Syringe

Syringe

Fitting (female)

Tube clamp


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Care and Maintenance55

5.1 Cleaning

Clean each component of this product if necessary. Traces of test fluid can harden and clog the needle so it is a goodtime to clean syringes by flushing a cleansing solution through them.

5.2 Parts Treatable in Autoclave Sterilization

All parts used in this product can be sterilized in an autoclave. However, silicon tubes, couplings, and fittings wear with

use and should therefore be replaced periodically.

After treating each part with an autoclave, dry it completely with an apparatus such as dryer (60C).

CAUTION

Caution in autoclaving

The heat applied when using an autoclave to sterilize bottle lids can cause the fitting to become loosened.

Check the fitting in the bottle lid after autoclaving and retighten if loose.

Remove the check valve clamps from the bottle holder before autoclaving.

Unlock the check valve clamps and tube clamps before autoclaving.


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Specifications66

Model BS-P1, BS-P2

Operating condition Temperature: 18 to 28 C (Note that temperature within the culture chamber should be

set at 5 degrees higher than that of the environment)

Transportation

condition (when the

product is packed)

Temperature: -20 to 60 C

Humidity: 90% RH maximum (no condensation)

M494E Biostation IM Perfusion Unit

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Transcript of M494E Biostation IM Perfusion Unit