Kontron SFM25 Instruction Manual

KONTRON

INSTRUMENTS

SFM

Instruction

manual

18

Instruction

manual

SFM

TABLE

OF

CONTENTS

0.

GENERAL INFORMATION

1.

INSTALLATION

l . l

Unpacking

2

1.2

Checking

the

Instrument ' s

serial number

2

1.3

Checking

the

accessories

2

1.4

Checking

the

mains

supply

3

1.5

Installation

4

1.5.1 Installation

of the

xenon

lamp

6

1.5.2

Lamp

adjustment

7

1.5.3

Removal

of the

xenon

lamp

8

1.5.4

Fitting

the

sample

compartment

accessories

9

1.5.5 Adjustment

of the

HPLC

flow

cell

10

1.5.6 Printer, Recorder

11

2.

INSTRUMENT

DESCRIPTION

2.1

Specifications

2

2.2

Construction

of the

Instrument

4

2 . 3

Keyboard

6

2.4

Display

8

2.5

Parameter plan

9

2.5.1

Wavelength

control (group

2) 11

2.5.2 Data control (group

3) 12

2.5.3

Printer/Plotter

(group

4) 14

2.5.4 Interface control

RS-232C

(group

5) 16

3.

OPERATION

3.1

Operating hints

2

3.2

Initialisation

2

3 . 3

Programming

3

3.3.1

General remarks

3

3.3.2 Wavelength

programming

4

3.3.3 Calibration

5

3.3.4 Blank

compensation

6

3.3.5

Calculation

of

average

value

7

3.3.6 Emission

scan

8

3.3.7 Exciation scan

10

3.3.8

Synchroscan

12

3.3.9 Parameter viewing

14

3.3.10

Printout

of

parameter

15

3.3.11

Automatic data Output

Lambda-FIX

16

3.3.12

Automatic data Output

Lambda-SCAN

17

3.4

Measurement

examples

18

3.4.1

Fixed

wavelength

18

3.4.2 Emission scan

21

3.4.3 Excitation scan

24

3.4.4 Synchroscan

27

3.5

Changing

the

spectral

bandwidth

30

3.6

Switching

off the

Instrument

31

07.86

0

GENERAL INFORMATION

- 2 -

Instruction

manual

SFM

T A

4.

4.1

6.

6.1

6.2

6.3

6.4

6.5

6.6

BLE

OF

CONTENTS

(cont.

)

ERROR

DETECTION

Error messages

OPTIONS
Thermostatted 4 cell

changer

for

10x10

mm

Standard cells

91-00431

HPLC

flow

cell with holder

91-00430

Slit

holder with slit

5 and 10 nm

91-00432

Slit

holder with slit

10 and 20 nm

91-00481

Filter

holder

for

excitation

and

emission

91-00434

Software extension pack

(Instr.

84-00095)

Plotter/Printer

and

method

files

91-00441

6.6.1

Storing a method

6.6.2 Recall

of a

method

file

6.6.3 Intallation instruction

6.6.4 Operation

6.6.5

P800

used

äs

plotter

in

TIME DRIVE

6.6.6

P800

used

äs

plotter

in

LAMBDA DRIVE

6.6.7

How to

produce

an

overlay

on the

P800

6.6.8

Automatic curve overlay with

P800

2

2

2

3

4
5
6

8

10
11

(for

SW

Version

8603

and

up)

6.6.9 Baseline correction

13

(for

SW

Version

8603

and up)

6.7

Photomultiplier

R446

(200-800

nm)

91-00433

6.8

Polarization

kit

(Instr.

84-00219) 91-00493

7.

APPENDIX

7.1

Optical diagram

2

7.2

Mechanical construction

3

s^

7.3

Block diagram

4

7.4

Software block

5

7.5

Sample cells

6

7.6

Function control

7

7.7

Communication

via RS 232 11

7.7.1 Format

of

data-out-string

11

7.7.2

Code

Table - Interface

RS 232 12

7.7.3

SFM 25

entry mask

13

07.86

0

GENERAL INFORMATION

- 3 -

K3

Instruction

manual

SFM

1.

INSTALLATION

l. l Unpacking

2

1.2

Checking

the

Instrument's

serial

number

2

1.3

Checking

the

accessories

2

1.4

Checking

the

mains supply

3

1.5

Installation

4

1.5.1 Installation

of the

xenon

lamp

6

1.5.2

Lamp

adjustment

7

1.5.3

Removal

of the

xenon lamp

8

1.5.4

Fitting

the

sample compartment accessories

9

1.5.5

Adjustment

of the

HPLC

flow

cell

10

1.5.6

Printer, Recorder

11

09.84

l

INSTALLATION

- l -

Instruction

manual

SFM

1.1

UNPACKING

Compare

the

consignment

with-

the-

delivery

note.

If

parts

are

missing

or

damaged,

please

inform

your

local

KONTRON representative immediately.

Transport

the

Instrument

only

in

its

original packing

box.

1.2

CHECKING

THE

INSTRUMENTS

SERIAL

NUMBER

The

plate

with

the

serial number

is

mounted

on the

left

side

of the

Instrument. This number

must

be

identical with

the

serial number

on the

delivery

note.

1.3

CHECKING

THE

ACCESSORIES

The

following

Standard accessories

are

included:

Number
of

items

1
1
1

1
1
1
1
1

Description

Mains

cable

Schuco

Recorder

connection cable

4

cell

changer (installed)

or

HPLC-flow

cell

Hex

wrench

size

3

Fuse

set

Xenon

lamp

150W

Instruction

manual

Installation Report

Order

no.

55-65601
93-00325
89-00200
91-00430
40-30003
91-00435
54-01001
95-00161
84-00046

07.86

l

INSTALLATION

- 2 -

Instruction

manual

SFM

1.4

CHECKING

THE

MAINS

SUPPLY

The SFM 25 is

factory

set for

220V

50/60HzL.-

For. Operation with other

mains

voltages,

the

correct voltage must

be set on the

voltage selector

(see

picture

below).

Possible

voltages:

100, 120,

220 und 240 VAC

The

fuses

on the

voltage selector

(1) and

XENON

lamp

(2)

have

to be

adjusted according

to the

mains supply.

Frequency

adjustment

for

50/60Hz

is not

necessary.

_7

WARNING

DISCQNNF.CT

FROM

MAINS

BEFORE

OPENiHQ

FUSES

VDLTAQE

HANGE

MOTECTXWfiEMlCE

war

WPH

TUE

S*WE

TYPE

AXQ

RITWO

Unplug

mains

cable

Open

the

snap-cover

(A)

with a screw

driver

at the top

Pull

marker

wheel

(B) and

turn

it to

the

required position

Push

(B)

back into position

Pull

out

fuse holders

(C)

insert

the
correct fuse
Push holders

(C)

back into position,

obey

arrows

Close

snap-cover

(A)

Check

if the

correct voltage

is

displayed

04.86

INSTALLATION

- 3 -

K<

Instruction

manual

SFM

For

your

notes:

09.83

l

INSTALLATION

- 4 -

Instruction

manual

SFM

1.5

INSTALLATION

The

Installation

site

of the

Instrument

should

be

such that

it is not

subject

to

dust

or

vapours

of

solvents

and

acids.

The

ambient

temperature should

be

between

10°C

and

35°C.

It

is

most

important

that there

is

sufficient free space

around

the

Instrument

for

adequate Ventilation.

air

outlet

mini

mum

30 cm

'

1

1

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FROM MAINS BEFORE OPENINQ

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09.83

l

INSTALLATION

- 5 -

Hü

Instruction

manual

SFM

1.5.1

INSTALLATION

OF THE

XENON

LAMP

ATTENTION:

The

xenon

lamp

is a

high pressure

lamp.

Handle

it

with
care.
EXPLOSION

HAZARD!

The

lamp protection cover

must

be

fitted before

working

on

or

around

the

lamp.

Wear a face protection shield

while

fitting

the

lamp

protection cover.

Procedures:

fS

-

disconnect

mains

cable

on the

Instrument

- to

remove

the

rear

panel

unscrew

the

four

screws

(A).

Screw

one

into

(B) and

remove

the

rear panel carefully

by

pulling

on the

screw

(B).

-

wear face protection

remove

the

lamp

housing

by

screwing

out the

screws

(C)

-

carefully screw

in the

lamp

with protective

cover,

anöde

(+) on the

top, into

the

lamp

holder

Turn

the

central screw

(D)

with

the hex

wrench

so

that

the

glass

ridge

(E) on the

lamp

points

to the

right. Perform a coarse

positioning

of the

lamp

with

help

of the

table

(chap.

1.5.2)

-

Wind

the

cathode connection

cable

(K)

around

the

lamp

and

with

the

knurled

nut

clamp

it

onto

the

cathode

(-).

-

Remove

carefully

the

lamp

protection cover

Screw down

the

cover

to the

lamp

housing.

Tighten

all 4

screws

(C)

equally.

-

Face protection shield

can be

removed

Remount

the

back panel with

the 4

screws

(A).

11.83

l

INSTALLATION

-

6 -

Instruction

manual

SFM

1.5.2

LAMP

ADJUSTMENT

Switch

on the

Instrument

and

wait about

10 min

Open

the

sample

compartment

cover

and

remove

the

accessory

(cell

holder,

LC-Cell

holder)

Adjust

the

excitation

wavelength

to 540

nm

Hold a white

paper

or

card

on the

lens

tube

in the

excitation

beam

The

lamp

should

be

adjusted

with

the

adjusting

screws

(G, H, I) so

that

the

highest

light

intensity

is

seen

on the

card.

The

possible adjustments

of the

lamp

holder

are

described

below.

Movement

of

the

lamp

1

down

1

up

v.

back

X~ f orward

»

right

-«

left

Screws

G,

H,

I

G,

H, I,

H

H

I

I

Direction

of

rotation

clockwise (all 3 equally)

counter clockwise (all 3 equally)

clockwise

counter clockwise

clockwise

counter clockwise

09.83

l

INSTALLATION

-

7 -

Instruction

manual

SFM

1.5.3

REMOVAL

OF

THE

XENON

LAMP

ATTENTION:

The

xenon

lamp

is a

high pressure

lamp.

Handle

it

with

care!

EXPLOSION

HAZARD!

The

lamp protection cover must

be

fitted before working

on

or

around

the

lamp.

Wear a face protection shield

while

fitting

the

lamp

protection cover.

Switch

the

Instrument

off at

least l hour prior

to the

lamp

removal.

Never exchange

hot

lamps.

Procedure:

-

Disconnect

mains

plug

on the

Instrument

- Let the

Instrument

cool down!

-

Remove

back

panel

(screws

A)

-

Use

face protection

-

Remove

cover

on the

lamp

housing (screws

C)

- Fit the

lamp protection cover around

the

lamp

-

Loosen

the

knurled

nut on the

cathode

(-) and

remove

the

cathode connection

cable

(never

try to

unplug)

-

Screw

the

lamp

out of the

lamp

holder

and put it

back

into

the

packing box.

The

Installation

of the

xenon

lamp

is

described

in

1.5.1.

09.83

l

INSTALLATION

-

8 -

3

Instruction manual

SFM

1.5.4

FITTING

THE

SAMPLE

COMPARTMENT

ACCESSORIES

The

SFM 25 is

delivered

with a cell

changer

or a

HPLC

flow

cell

The

sample compartment accessory

is

placed

into

the

sample

compartment

and

fastened

with

the

fixing screw

F.

The

front

plate

P has an

easy removable snap

fixing.

Through

the

tube

guide,

the

necessary cables

or

tubes

can

easily

be

led

into

the

sample

compartment.

09.84

I

INSTALLATION

-

9 -

•B

Instruction manual

SFM

1.5.5

ADJUSTMENT

OF THE

HPLC

FLOW

GELL

Flow

direction

out

Flow

direction

in

max.

operating pressure

: 50 bar

The

flow

cell

has

been mounted

and

adjusted

in the

factory.

A re-

adjustment

is not

required.

If

there

are

difficulties

while

mounting

the

modul

or

when

in

use, then

please

contact your KONTRON

representative.

09.84

INSTALLATION

- 10 -

Kl

Instruction manual

SFM

1.5.6

CONNECTION

OF

ACCESSORIES

A

variety

of

peripherial equipment

can be

connected

to the SFM 25

e.g.:

Recorder (Y/t

and

X/Y),

Printer,

Printer/Plotter

or

Computer.

The

following

plugs

are

already

prepared

on the

rear

panel.

l.

Analogue

out-

put

for

Recor-

der

and

inte-

grator

2.

RS-232C

for

Printer,

Prin-

ter

/Plotter

or

Computer

Recorder

cable

The

recorder cable

can be

plugged

in, on the

15-pol

socket

(1)

Following signal

are

available:

REM

violet (Remote

ON/OFF)

GND

yellow

REG

grey

GND

pink

A red

GND

black

Shield

blue

INT

GND

white
brown

-a

Shield green

Not

used cable ends

are to be

insulated.

Pin

assignment

of RS

232C-Interface

(Socket

2)

Recorder

Integrator

GND
DSR

CTS
RxD

DTR

RTS

TxD"

Also

see

appendix, chapter

7.7

09.84

l

INSTALLATION

- 11 -

Ifl

Instruction manual

SFM

2.

INSTRUMENT

DESCRIPTION

2.1

Specifications

2

2.2

Construction

of the

Instrument

4

2.3

Keyboard

6

2.4

Display

8

2.5

Parameter

plan

9

2.5.1

Wavelength

control (group

2) 11

2.5.2 Data control (group

3) 12

2.5.3

Printer/Flotter

(group

4) 14

2.5.4

Interface control

RS-232C

(group

5) 16

04.84

2

INSTRUMENT

DESCRIPTION

- l -

Instruction

manual

SFM

The

SFM 25 is a

high performance Spectralfluorometer

to

measure

in

the

wavelength

ränge

of 200 - 800

nm.

It

is

controlled

by 3

microprocessors

which

allow

simple

Operation over

the

keyboard

or

complete external

control.

The

flexible

conception

of the

Instrument

gives

numerous

expansion

possibilities.

Both

the

excitation-

and

emission

monochromator

are

equipped with holographic

stigmatic

corrected blazed concave

gratings

in a

Seya-Nomioka-arrangement.

The

spectral

bandwidth

of the

monochromators

is

switchable

from

10 to 15 nm.

Slits

with

5 and 10 nm

bandwidth

are

available.

The

light

source

is a

150W

xenon

high

pressure

lamp.

The

lamp

house

is

sealed.

The

Standard version includes a reference beam System. This provides high

signal

stability even with temporary

energy

variations

of the

lamp caused

by

movement

of the

are,

shimmering

of the air

surrounding

the

lamp

or

other

thermal

effects.

The

reference detector

is a

photo diode.

The

light emitted

by

the

sample

is

measured

with a photomultiplier.

The

accessories such

äs 4 cell

changer,

thermostatted cell holders, HPLC

flow

cell

are

built

on a

modular

basis

and are

easely fitted into

the

sample

compartment.

2.1

SPECIFICATIONS

Optical

System

Wavelength

ränge

Monochromators Excitation

and

Emission

Excitation
Emission

Band

width

Wavelength

accuracy

Wavelength

precision

Lamp

power supply

Light

sources

Lamp

housing

high energy

throughput

double
monochromator System with reference
beam

compensation

of

source Output

changes

200 - 800 nm

Concave holographic gratings,
1200

line/mm;

f = 100 mm, F

3.5;

lineare dispersion

= 8

nm/mm

Blazed

at 250 nm

Blazed

at 450 nm

10 nm, 15 nm, 20 nm

selectable

(5 nm

options)

1.0 nm

0.1 nm

switching regulator, power Output
regulated

Xenon-High pressure

lamp

150W

sealed

10.84

2

INSTRUMENT DESCRIPTION

-

2 -

ä

Instruction

manual

SFM

2.1

SPECIFICATIONS

Sample

detector

Reference detector

Fluorescence display

Display

Scan speed

Scan ränge

High

voltage

Calibration

Blank compensation

Response

time

Sensitivity

Photomultiplier

R 212

Range

200-650

nm

(Option: Photomultiplier

R446

Range

200-800

nm)

Photo diode

0...+

200.0

alphanumeric;

16-digit

20-500

nm/min

in

steps

of 5

nm/min

1-600

nm in

steps

of l nm

100-999

volts

automatically

selected

by

the

microprocessor

automatic

automatic

0.5,

l, 2, 4, and 8

sec.

for

98% of

füll

scale

Raman

band

of

water

Ex

350 nm; Em 397 nm;

slit

10 nm;
Response time 8 sec;
S/N

better

than

70:1

Analog Outputs:

Fluorescence

Integrator
Wavelength
Remote

Digital Interface

Power

requirements

Dimensions

Weight

Printer/Plotter

100 mV for 200 %

rel.

fluorescence

IV

for 200 %

rel.

fluorescence

100 mV /

1000

nm

TTL - Signal

(5V)

for

Start/Stop

RS-232C

bidirectional

Baudrate

110-2400

Format

5-8

bits

Parity

odd,

even,

off

Stop bits

l or 2

100,

120, 220,

240 VAC

50 / 60

Hz;

400 VA

275 mm

high;

532 mm

wide;

517 nun

deep

37 kg

Option

07.86

2

INSTRUMENT DESCRIPTION

- 3 -

SS

Instruction

manual

SFM

2.2

CONSTRUCTION

OF THE

INSTRUMENT

Parameter plan

Display

field

Keyboard

Sample

compartment

Power ON/OFF

09.84

2

INSTRUMENT DESCRIPTION

-

4 -

i

Instruction

manual

SFM

2.2

CONSTRUCTION

OF THE

INSTRUMENT

@©

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WARNING

DISCONNECT

FROM

MAINS

BEFORE OPENINQ

FUSES

HAINS

4A
IA

©

VOLIAUE

MNGE

loo-mv
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l)

Analog Output

2}

RS-232C - Interface

3)

Mains

inlet

with selector

and

fuses

4)

Earth

terminal

5)

Fuses

for

lamp

power supply (xenon

unit]

6) Air

outlet

7)

Cover

plate

memory

board

8)

Cover plate

for

spare places

10.84

2

INSTRUMENT DESCRIPTION

- 5 -

'•[i

Instruction

manual

SFM

2.3

KEYBOARD

The

keys

are

divided

into 2 groups:

a)

Keys

for

numerical

entries

clear

return

Clears

the

last

number

entered

Resets

errors

or

error

messages (see chapter

4)

AUTO

ZERO

Stores

the

entered number

and

returns

to the

measuring

mode

Stores

the

entered number

and

advances

to the

next

line

to

enter numerical values

and

ON/OFF states

b)

Function

keys

A

T

A-Search:

These

keys

are

used

to

move

the

excitation

or

emission

monochromator

manually.

The

monochromator

can be

moved

in

single steps

by

short, repeated pressing

on the

key.

For

continuous

movement

keep

the key

pressed.

The

monochromator speed during

the

first 4 sec.

is 145

nm/min

and

then

930

nm/min.

The

monochromator

moves

back

to the

original

value

after

pushing

the

"return"

key.

Automatic

calibration

After pressing

the key the

Instrument

sets

all

para-

meter

values

so,

that

the

measured

fluorescence

intensity corresponds with

the

reference value under

mmm

CALIBRATION.

The

high voltage

on the

photomulti-

plier

is

regulated

and the

value stored under

•(*••'•

HIGH

VOLTAGE,

MM

BLANK

and

•«•

FACTOR

are set to

0 and 1.0

respectively.

10.85

2

INSTRUMENT

DESCRIPTION

-

6 -

.

fei

Instruction

manual

SFM

2.3

KEYBOARD

b)

Function

keys

(continuation)

Automatic

blank compensation

The

current fluorescence

value

is

measured

and

stored under

•CHCB

BLANK. This value

is

subtracted from

all

following

measurements.

In

addition

the

Instrument

automatically

calculates

the

necessary factor

which

is

required

to

reach

the

programmed reference value

in

»ciüM

CALIBRATION,

and

stores

it

under

BEIEB

FACTOR.

This procedures

ensures

that

the

fluorescence intensity

of the

reference substance

is

used

äs

Standard. This function will

not be

carried

out,

if

the

blank value exceeds

90% of the

entire fluorescence

intensity

of the

reference substance.

Auto

zero

The

current

fluorescence

value

is

measured

and

stored under

•elica

BLANK. This value

is

subtracted from

all

following

measurements.

The

9&!M

FACTOR remains unchanged.

Calculation

of the

average value

Output

on

RS-232C

Interface

This

key

activates a calculation

of the

average value

of the

relative

fluorescence.

The

time

base

is the

preprogrammed

value

in

BtlÜM

RESPONSE

TIME.

The

fluorescence intensity

is

averaged

during this time.

The

mean

value appears

on the

display

for 2

sec.

[MEAN

101.

s]

This value

is

Output

to the

RS-232C

Interface.

The

analog

Signals (recorder,

Integrator)

are not

influenced

by

this

function.

Start/Stop

of

scan

This

key

controls

the run of a

scan programmed

in

parameter

group

2. The

procedure

is

only

performed

if the

sample

compartment

cover

is

closed.

10.85

2

INSTRUMENT DESCRIPTION

-

7 -

Iß

Instruction manual

SFM

2.4

DISPLAY

PANEL

STATE

DISPLAY

MEASURE­MENT

MODE

-Fix

4 n n

i

f

JU

tl

£-

relative

fluorescence

nm

emission

current

wavelength

of the

excitation

mono-

chromator

value

of the
relative
fluorescence

current
wavelength

of the

emission

mono-

chromator

A—Scan

(EX,

EM,

EX

+ EM)

display

of a

.A-Scan-mode

A

—Scan

Stop

Display

of an

Interruption

of a

scan

Parameter­Mode

Display

of the

selected parameter with

its

value

Error­Mode

alphanumerical display

of an

error

09.83

2

INSTRUMENT DESCRIPTION

-

8 -

K<

Instruction

manual

SFM

2.5

PARAMETER PLAN

The

parameter plan

is

divided

into 9 groups.

Each group

can be

called

up

via the

related

number.

Stepping through

all

parameters

in the

selected

group,

is

done

by

repeated pressing

of the

"enter"-key.

The

values

may be

changed

within

the

allowed

ranges.

The

groups

l, 2 and

3 are

displayed

more

detailed

for

easier

handling.

Single parameters

are

selectable directly.

Example:

Selection

of

parameter

EMISSION

SCAN

There

are

two

ways

to

select

the

parameter

Hüll

EMISSION

SCAN:

1.

2.

EM.SCAN

ON

[EM.SCAN

ON

Parameter groups

t

t

to

fill

out

by

the

user

-

File name

-

Code

-

etc.

•preprogrammed

values

•allowed

ränge

parameter

09.84

2

INSTRUMENT

DESCRIPTION

-

9 -

-S

Instruction manual

SFM

For

your notes:

09.83

2

INSTRUMENT

DESCRIPTION

- 10 -

18

Instruction manual

SFM

2.5.1 WAVELENGTH CONTROL (group

2)

EXCITATION

: - The

excitation

wavelength

can be

selected

in the

ränge

200 to 800 nm, in

steps

of l

nm.

The

programmed

value

is

also

the

starting

wavelength

for an

excitation scan. Value

0 is

also

allowed,

then

the

monochromotor

will

be set

to

zero

order.

This

means

that

the

sample

is

irradiated with white

light

from

the

lamp.

EMISSION

: - The

excitation wavelength

can be

selected

in the

ränge

of 200 to 800 nm, in

steps

of l nm.

The

programmed value

is

also

the

starting

wavelength

of an

emission scan. Additionally

value

0 is

allowed, then

the

monochromator

is

set to

zero order. This means,

all

emitted

ligth

is

transmitted

to the

photomultiplier.

EXCITATION:

-

This parameter

is

programmed with

"ON"

or

"OFF".

SCAN

It

states whether

the

excitation monochromator

is

moving

or not

while

performing a scan.

EMISSION

SCAN: - This parameter

is

programmed with "ON"

or

"OFF".

It

states whether

the

emission monochromator

is

moving

or not

while performing a scan.

For

Synchroscan

it is

necessary

to

program both

and

tat!EMISSION

SCAN with "ON".

EXCITATION

SCAN

SCAN

RANGE:

- The

wavelength ränge

is

selectable

in the

ränge

of l to 600 nm in

steps

of

Inm.

The

value

for the

start wavelength

is

preprogrammed

in

EB£|

EXCITATION

and

ME»

EMISSION.

The

ränge i s

scanned from

the

long

to

short

wavelength.

SCAN

SPEED:

- The

scan speed

of the

monochromator

is

selectable

between

20 and 500

nm/min,

in

steps

of 5

nm/min.

If a multiple

of 5nm is not

programmed,

then

the

Instrument

rounds

to the

closest value

automatically.

09.83

2

INSTRUMENT DESCRIPTION

- 11 -

IS

Instruction manual

SFM

2.5.2 DATA

CONTROL

(Group

3)

CALIBRATION:

The

value

programmed

under this parameter

is

the

reference value

for the

calibration. After

pushing

the

"calib"

key.

The

Instrument

regu-

lates

the

high voltage

on the

photomultiplier,

to

match

the

measured fluorescence

with

the

reference value.

The

ränge

of

this value

is 0.1

to

200.0

HIGH

VOLTAGE

LEVEL

This

parameter

is a

scale

for the

high voltage

on

the

photomultiplier.

The

"calib" function

automatically

sets

the

value

of the

voltage.

It

can

be

programmed

manually

in the

ränge

from

100-999.

This value

is

approximately

the

high

voltage

in

volts.

BLANK:

The

blank value

is

subtracted from

the

measured

fluorescence.

The

blank value

is

measured

and

stored

under this parameter

whenan

"auto

blank"

or

"auto

zero"

is

pressed. Programming

via the

keyboard

in the

ränge from

0.0 to

800.0

is

also
possible.
MM

FACTOR

is

always

reset

to 1.0

when a new

blank

is

entered

or

measured.

FACTOR:

The

FACTOR

is

used

to

multiply

the

measured

fluorescence.

The

value

is

automatically

calculated

and

stored under this parameter when

"auto blank"

is

pressed.

The

factor

can

also

be

entered

manually

in the

ränge from

0.01

to

10.0

Both

parameters

mentioned above

are

used

in the

calculation

of the

relative

fluorescence

äs

follows:

(S-B)

x F = RF

S =

measured fluorescence intensity

B

=

programmed

blank value

BeHeM

BLANK

F =

programmed factor

mtf^m

FACTOR

RF = displayed relative fluorescence

10.85

2

INSTRUMENT DESCRIPTION

- 12 -

Instruction

manual

SFM

2.5.2 DATA

CONTROL

(Group

3)

RESPONSE:

This parameter

is

used

to

control

the

speed

of

response

of the

display

and the

analog output

(recorder

and

Integrator).

The

"response

time"

is

defined

äs

the

time

required

to

reach

98% of the

füll

scale

during

fast

signal

changes.

The

following

time

constants

are

programmable:

0.5,

l,

2, 4, 8

sec.

RATIO:

^

SELF

CHECK:

Switching

on and off of the

ratio System

is

performed

with

this parameter

by

programming

with "ON"

and

"OFF".

The

built

in

ratio

System

compensates

for

changes

in the

lamp

Output.

This parameter

performs a self-check,

but

without a display check.

All

parameters

of

groups

2 to 5 are

reset

to the

preset

values.

09.83

2

INSTRUMENT DESCRIPTION

- 13 -

Iß

Instruction manual

SFM

The

following

parameter

groups

are not

displayed

in

detail

on the

parameter

plan.

The

single parameter appear

after

selection

of the

group

on the

display.

The

programming

is

performed

äs

for

groups

2 and 3.

2.5.3

PRINTER/PLOTTER

(Group

4)

This block contains

all

data

for

printing

out the

current

parameters.

CODE:

Entering

of a

10-digit

code

or a

8—digit

date,

Example:

CODE

or

0467213598

12.03.1984

Code

or

date appears

in the

heading

of the

parameter

list.

LISTING:

As

soon

äs

this

parameter

is

programmed with

an

"ON",

the

parameter listing

is

sent

to the

RS-232C

Interface.

ATIME

SEC

This function allows printout,

via the

RS-232C,

of

relative

fluorescence

äs a function

of

time.

It

is

enabled

by

pressing

"ON".

The

programmed

value

under

mwtsm

DATA RATE

is a

time interval

of

seconds.

It is

activated

by the

start/stop

key.

ALAMBDA

NM

This function allows printout,

via the

RS-232C,

of

relative

fluorescence

äs a function

of

wavelength.

It

is

activated,

in the

scan

mode

only,

by re-

pressing

"ON".

The

integer part

of the

value

programmed

unter

mwvm

DATA RATE

is a nm

internal.

(0 to l nm is

read

äs l nm)

DATA

RATE

The

measurement

internal

for the

above

two

para-

meters

is

enterd

here.

Depending upon

the

mode

selected

the

value

is in

seconds

or nm.

Parameters

mmtm

ATIME

SEC and

BICT

ALAMBDA

NM can not be

activated

at

the

same time.

When

one of

these parameters

is

switched

on, the

Instru-

ment

automatically

switches

the

other

off.

09.84

2

INSTRUMENT DESCRIPTION

- 14 -

9

Instruction manual

SFM

2.5.3 PRINTER/PLOTTER

(Group

4)

The

limitation

of the

data Output rates depend upon

the

Baudrate,

a)

Time dependent Output

Baudrate

2400,

4800

1200

600

300

150

110

smallest

time

interval

0

0

0

0

1

2

.1

s

.2

s

.4

s

.8

s

.6

s

.2

s

b)

Wavelength

dependent Output

Baudrate

2400,

4800

1200

600

300

150

110

fastest

scan speed possible

in

nm/min

with intervals

of

:

20

nm

500

500

500

500

500

500

10 nm

500

500

500

500

350

250

5 nm

500

500

500

350

150

125

2 nm

500

500

250

125

65

50

1 nm

500

250

125

65

35

25

If

these limits

are

neclected

on

programrning

of the

Instrument,

then

the

scan

is set to the

nearest interval during

start

procedure.

09.84

2

INSTRUMENT DESCRIPTION

- 15 -

KK

Instruction

manual

SFM

2.5.4

INTERFACE

CONTROL

RS-232C

(Group

5)

The

Standard

SFM 25 is

equiped

with a serial

RS-232C

Interface,

necessary

parameters

are

programmed

in the

group INTERFACE

CONTROL.

The

RS

BAUDRATE:

Following

baudrates

are

selectable:

110,

150, 300, 600, 1200, 2400, 4800

Baud

A

complete

data

package

for a

measurement

signal

with

the

wavelength

for

excitation

and

emission requires

22

characters.

These

are

transmitted

via

RS-232C

to a

periphery.

Example:

with a Baudrate

of

1200

baud

(assuming 8 data bits

+ 2

stop bits

per

character)

1200

22 x 10

=

5,4

measurement point

per

sec.

can be

transferred.

The

baudrate

has

to be

adjusted

to

correspond with

the

pheripheries (Computer, Printer, Plotter

etc.).

RS

DATA FORMAT:

5, 6, 7 or 8

Data bits

per

byte

are

possible.

RS

PARITY:

0 =

without parity

1

= odd

parity

2 =

even parity

DATA

FORMAT

maximum 7 Bits

RS

STOP BITS: Data bytes with

l or 2

stop bits

can be

selected.

04.84

2

INSTRUMENT DESCRIPTION

- 16 -

KÜ

Instruction

manual

SFM

3.

OPERATION

3.1

Operating

hints

2

3.2

Initialisation

2

3.3

Programming

3

3.3.1 General remarks

3

3.3.2

Wavelength

programming

4

3.3.3 Calibration

5

3.3.4 Blank

compensation

6

3.3.5 Calculation

of

average

value

7

3.3.6 Emission scan

8

3.3.7 Excitation scan

10

3.3.8 Synchroscan

12

3.3.9 Parameter

viewing

14

3.3.10

Printout

of

parameters

15

3.3.11

Automatic

data

Output

A-FIX

16

3.3.12

Automatic data Output A-SCAN

17

3.4

Measurement

Examples

3.4.1

Fixed

wavelength

18

3.4.2 Emission scan

21

3.4.3 Excitation scan

24

3.4.4 Synchroscan

27

3.5

Changing

the

spectral

bandwidth

30

3.6

Switching

off the

Instrument

31

10.85

3

OPERATION

- l -

Kü

Instruction manual

SFM

3.1

OPERATING

HINTS

The

SFM 25

must

be

installed according

to the

Installation instruction

(chapter

1).

AVOID

REPEATED

SWITCHING

ON AND OFF IN

ORDER

TO

PREVENT

DAMAGE

TO THE

XENON

LAMP

AND

ELECTRONICS

3.2

INITIALISATION

SW-VERSION

XXXX

)

—

power

on

—

Automatic

ignition

of the

xenon

lamp

i

[

SELF CHECK

~)

l

The

actions:

Initialisation

program takes 3 min.

and

performs

following

ms:

-

Display test

-

Display

of the

actual Software Version

-

Initialiation

of the

System

-

A-Positioning

-

Defines

parameter

values

-

Seif

check

of

motors

and

motor

control,

-

Seif

check

of

AD-DA Converter

-

Seif

check

of CPU +

memory

-

Seif

check

of

data

transfer

l

On

the

display

appears:

|

[

480 0.0 480

J

The

Instrument

is

ready

to

operate.

The

lamp

energy

is

stable after

10

min.

09.83

3

OPERATION

- 2 -

ß

Instruction manual

SFM

3.3

PROGRAMMING

The

SFM 25 has 4

measuring

modes:

-

Fixed wavelength

-

Emission scan

-

Excitation

scan

-

Synchroscan

(simultaneous

scanning

of Ex— and Em-

monochromators)

Each

mode

of

parameter

programming

is

described with

an

example

in

chapter 3.4.

Key

functions,

parameters

and

displays

are

described

in

chapter

2.

The

ränge

of

each parameter

is

described

on the

parameter plan

and in

chapter

2.

3.3.1

GENERAL REMARKS

After

power

on a

test program

is

performed.

When

the

test

is

completed

the

parameter values

are

programmed with

the

preset values according

to

the

parameter plan.

On

the

display appears

[

480 0.0 480

J

Parameter changes

can

now

be

made.

The

limits

of the

parameters

are

controlled

by the

microprocessor.

09.83

3

OPERATION

- 3 -

Kl

Instruction

manual

SFM

3.3.2

WAVELENGTH

PROGRAMMING

Example

of a

wavelength

adjustment:

EXCITATION

350 nm

EMISSION

520 nm

[ 480

fff.f

480 )

[

EXCITATION

480 )

5

—

2

—

0

OFF

[350

fff.f

520]

Instrument

display

after

Initialisation

fff.f

denotes

any

fluorescence

selection

of

parameter

group

2

"WAVELENGTH-CONTROL"

-

display

of

excitation wavelength

-

enter

excitation wavelength required

[

EXCITATION

350]

-

display

of

value

entered

storing

of

last value entered

and

advance

to

next parameter

[

EMISSION

480 ] -

display

of

emission wavelength

-

enter

the

emission wavelength required

[

EMISSION

520J

-

display

of

value entered

storing

of the

value entered

and

return

to

the

measuring

mode

All

parameters

can be

changed

äs

described above.

09.83

3

OPERATION

-

4 -

IS

Instruction manual

SFM

3.3.3

CALIBRATION

The

SFM 25 can be

set-up

for the

required working ränge

with

the

"calib"-key.

Example:

Calibration

to 50%

relative

fluorescence

Set the

wavelengths

to the

required

values;

e.g

Raman-band

of

water

is

Ex

= 350 nm /

Em

= 397 nm

[350

fff.f

397]

-

fff.f

= any

fluorescence

[CALIBRATION

100.0]

l

[

CALIBRATION

50 )

{ 350

fff.f

397]

[

CALIBRATION

RUN

j

[350

50.0

397]

selection

of the

parameter

CALIBRATION

-

display

of

calibration

value

-

enter calibration value required

-

display

of

value entered

-

storing

of the

value entered

and

return

to

the

measuring

mode

- put a

cell

containing bidistilled water

into

the

cell holder

and

close

the

sample

compartment cover

-

press

key

"calib"

- The

calibration

is

carried

out;

after

approx. 4 sec.

the

Instrument

automatical-

ly

Switches back into

the

measuring

mode.

In

order

to

carry

out the

calibration

äs

quickly

äs

possible with

äs

many

data points

äs

possible,

the

shortest available

time

constant

is

used. Therefore with very noisy Signals

it

may

be

necessary

to

press

"calib" a second time.

09.83

3

OPERATION

- 5 -

KB

Instruction

manual

SFM

3.3.4

BLANK

COMPENSATION

Automatical

compensation

for a

blank

value.

Example:

Calibrate

Instrument

0

[ 480 0.0

570J

put a cell

containing blank solution into

the

cell holder

[480

fff.f

570J

-

display

of

blank value

-

start

of

automatic blank compensation

The

blank value measured

is

automatically

stored under

•tue»

BLANK.

This

blank value

is

subtracted

from

all

subsequent

measurements.

09.83

3

OPERATION

-

6 -

H

Instruction manual

SFM

3.3.5

CALCULATION

OF

AVERAGE

VALUE

The

"read/print"

key

allows

an

average relative fluorescence

value

to

be

calculated.

The

value

programmed

under

•«•••

RESPONSE

TIME

is

used

äs

the

time

base.

The

relative fluorescence

is

averaged during this

time.

The

average value appears

on the

display

for 2

sec.

The

average

value

is

also available

at the

RS-232C

output.

The

analog signal

(recorder,

Integrator)

is not

affected

by

this process.

[

350

98.5

397

]

[

MEAN - COMPUTE

[MEAN

98.7

]

[350

98.8

397

J

—

display

in

measuring

mode

-

start

of

averaging

-

start

of

calculation

of the

average

value

-

display

of

result

for 2

sec.

-

display

of

measuring

mode

09.83

3

OPERATION

- 7 -

3

Instruction manual

SFM

3.3.6 EMISSION

SCAN

The

SFM 25

scan Operation with

the

emission monochromator

is

programmed

via the

parameter

HEB

EMISSION

SCAN. Necessary Parameters

such

äs

BSB3

SCAN RANGE

and

WS&M

SCAN

SPEED

have

to be

programmed

also.

Example:

Raman

band

of

water

Aex

350 /

Aem

397

Put a cell

containing bidistilled water into

the

cell

holder

and

calibrate

the

Instrument

at

AEx

350 / AEm

397.

l

350

100.0

397

j

2

•

4

enter

[

EM.

SCAN

OFF

]

1

ON

enter

[

SCAN

RANGE

250 )

-

select parameter

EMISSION

SCAN

preparing

the

emission monochromator

for

scan Operation

0

OFF

0

OFT

enter

[

SCAN

SPEED

100 J

-

enter scan ränge

200

nm

enter

<£

enter

-

scan speed o.k.

[EMISSION

397J

OFF

OFF

return

[350

fff.f

500 J

l

[ 350

fff.f

* 483 J

enter

Start wavelength (500

nm) and

return

to the

measuring

mode

-

start

of

emission scan

—

display during a scan

09.83

3

OPERATION

-

8 -

H

Instruction manual

SFM

3.3.6

EMISSION

SCAN

(continuation)

[ 350 *

fff.f*

470 )

[

350

fff.f

* 453

[350

fff.f

300]

scan

can be

interrupted

at

any

time

with

the

"start/stop"-key

-

display

in a

"scan stop"

mode

-

continue scan

- end of

scan

[ 350

fff.f

500 )

The

start

wavelength

can be

reset with

the

"return" key.

The

scanning

can be

terminated

at any

time

with

the key

sequence:

^

09.83

3

OPERATION

-

9 -

-8

Instruction manual

SFM

3.3.7

EXCITATION

SCAN

The

SFM 25

scan Operation

withthe

excitation monochromator

is

programmed

via the

Parameter

•8MM

EXCITATION

SCAN.

Necessary

Parameters such

äs

EB9

SCAN RANGE

and

•£&••

SCAN

SPEED

have

to be

programmed

also.

Example:

Calibrate

the

Instrument

with

the

sample

to be

measured

at a

wavelength

pair

(Ex and EM)

with

maximum

energy.

[342

150.0

482

J

2

«

3

[ EX.

SCAN

OFF]

J

[

SCAN

RANGE

250J

0

OFF

0

OFF

enter

[SCAN

SPEED

looj

l

3

8

0

J3FF

l

[

EXCITATION

342 )

[ 380

fff.f

482]

Start

stop

l

[

370

fff.f

* 482 )

-

select parameter

EXCITATION

SCAN

preparing

the

excitation monochromator

for

scan Operation

and

step

to

scan

ränge

-

enter scan ränge required

-

scan speed

o.k.

enter start wavelength

and

return

to

the

measuring

mode

-

start

of

excitation scan

-

display during a scan

09.83

3

OPERATION

- 10 -

Kü

Instruction

manual

SFM

3.3.7 EXCITATION SCAN

(continuation)

( 365

*fff.f

* 482 J

scan

can be

interrupted

at any

time

with

the

"start/stop"

key

-

display

in

"scan stop"

mode

l

(350

fff.f

* 482 J

[280

fff.f

482 J

[ 380

fff.f

482J

-

continue scan

- end of

scan

the

start

wavelength

can be

reset

with

the

"return"

key

The

scanning

can be

terminated

at any

time with

the key

sequence:

09.83

3

OPERATION

- 11 -

19

Instruction manual

SFM

3.3.8

SYNCHROSCAN

It

is

possible with

the SFM 25 to

work

with

both monochromators

at the

same

time.

Parameter

BEI

EXCITATION

SCAN

and

BEI

EMISSION

SCAN

have

to be

programmed

with

ON for

this

mode.

Parameter

•*••»•

SCAN

RANGE

and

EOJSCAN

SPEED

are

valid

for

both

monochromators.

Example:

Calibrate

Instrument

with

the

sample

to be

measured

at a

wavelength

pair

(Ex and Em)

with

maximum

energy.

[350

130.0

400j

2

enter

l

[

EXCITATION

350J

5

0

OFF

0

OFF

enter

l

[

EMISSION

400J

l

5

5

0

OFF

[

EX.

SCAN

OFF

1

ON

enter

[EM.

SCAN

OFF]

-

select

parameter

group

2

enter scan

start

wavelength

for

excitation monochromator

enter scan start wavelength

for

emission

monochromator

preparing both monochromator

for

scan

Operation

1

ON

enter

[

SCAN

RANGE

250J

l

3

0

OFF

0

OFF

enter

[SCAN

SPEED

lOOJ

-

enter scan ränge required

enter scan speed required

and

return

to

the

measuring

mode

09.83

3

OPERATION

- 12 -

1-3

Instruction

manual

SFM

3.3.8

SYNCHROSCAN

(continuation)

[ 500

fff.f

550 j

-

start

of

synchroscan

[480

fff

.f *

530J

l

l

[430

»fff.f * 480J

-

display

during scan

-

scan

can be

interrupted

at any

time

with

the key

"start/stop"

-

display

in

"scan stop"

mode

l

[420

fff.f * 470J

[200

fff.f

250 J

-

continue scan

- end of

scan

return

|

500

fff.f

550

the

start

wavelength

can be

reset with

the

"return"

The

scanning

can be

terminated

at any

time

with

the key

sequence:

09.83

3

OPERATION

- 13 -

Kü

Instruction

manual

SFM

3.3.9

PARAMETER

VIEWING

During a scan each parameter

can be

selected

and

shown

on the

display.

But

change

of

parameter values

are not

possible

in

this

mode.

The

display

is

reset

to the

measuring mode

after 3 sec.

Example:

(400

fff.f

* 493 J

3

•

2

enter

(HIGH

VOLTAGE

soo

J

[400

fff.f

* 490 J

-

after 3 sec.

09.83

3

OPERATION

- 14 -

1-8

Instruction manual

SFM

3.3.10

PRINTOUT

OF

PARAMETERS

A

complete

parameter

listing

of

group

2 and 3 can be

obtained

via the

RS-232C

Interface

on the SFM 25.

Therefore,

if a

printer

is

connected,

a

printout

of

each

set of

operating Parameters

can be

obtained.

Example:

(480

. 0.0 480

"]

CODE

-

selection

of

parameter group

4

7

•

0

OFF

8

3

[LISTING

OFF j

~

LISTING

RUN

]

enter a max.

10-digit

code

(e.g

date).

This input

is

optional,

a

printout

without code

is

also possible.

-

start

of

parameter listing

-

display during a listing

[LISTING

OFF j

- end of

listing

-

return

to

measuring

mode

03.84

3

OPERATION

- 15 -

Hü

Instruction

manual

SFM

3.3.11

AUTOMATIC

DATA

OUTPUT

A-FIX

An

automatic output

of

data

via the

RS-232C

Interface

with selectable

time

interval

is

programmed

in

parameter group

4.

Example:

[480

0.0

480

J

4

•

3

enter

[

TIME

SEC OFF

~~]

-

select paramter

ATIME

SEC

1

OH

•- — •

enter

——

enter

[DATA

RATE

i.o

]

[480

0.0

480

]

activate output routine

and

advance

to

RATE

enter time interval

in

sec,

return

to

measuring

mode

and

start

printout

[480

0.0 T 480 J

-

data

is

output every

10

sec.

- T

indicates TIME DRIVE

is on.

To

interrrupt

the

data output press

start/stop

key.

03.84

3

OPERATION

- 16 -

fö

Instruction

manual

SFM

3.3.12

AUTOMATIC

DATA

OUTPUT

The

SFM 25

prints

out the

measurement values

via the

RS-232C

Interface,

during a scan.

The

interval

is

selectable.

Digitalisation

of

spectra

is

simplified

by

this

method.

Example:

[480

0.0

480j

4

•

4

enter

[ALAMBDA

NM

OFF)

l

[DATA

RATE

i.o]

-

select

parameter

EKIALAMBDA

NM

-

activate output routine

[480

0.0

480]

enter

wavelength

interval

and

return

to

measuring

mode

The

Output

is

obtained after start

of a

scan,

and is

interrupted

by

pressing

"start/stop"

or at the end of a

scan.

Switching

off of

data Output

-

select parameter

mirtim

ALAMBDA

NM

-

switch

off

data Output

and

return

to

measuring

mode

09.83

3

OPERATION

- 17 -

H

Instruction

manual

SFM

3.4

MEASUREMENT

EXAMPLES

3.4.1

FIXED

WAVELENGTH

(with

calibration

and

blank)

Example

with

following

parameters:

EXCITATION

EMISSION

CALIBRATION

RESPONSE

350

nm

420 nm

50

2.0

( 480 0.0 480 J

2

enter

[

EXCITATION

480

-

display

after

Initialisation

-

select parameter group

2

display

of

current excitation

wavelength

-

enter

new

value

[

EXCITATION

350 J

[

EMISSION

480]

[EX.SCAN

ON J

-

display

of

value entered

störe

new

value

and

advance

to

next parameter

display

of

current

emission

wavelength

enter

new

value, störe

and

advance

to

next parameter

-

display

of

parameter

0

OFF

enter

switch

off

EX-SCAN störe

and

advance

to

next parameter

09.83

3

OPERATION

- 18 -

Hü

Instruction

manual

SFM

3.4.1

FIXED

WAVELENGTH

(continuation)

(with

calibration

and

blank)

-

EM.

SCAN

ON

1

\

OFF

retum

350

0.0 420 ]

1

3

enter

CALIBRATION

100 J

-

display

of

parameter

-

switch

off

EM-SCAN, störe

and

return

to

measuring

mode.

-

display

-

selection

of

parameter group

3

-

display

of

first parameter

HIGH VOLTAGE

400 ]

[RESPONSE

i.o

J

l

[RATIO

OFF J

RATIO

ON

Enter

new

calibration

value,

störe

and

advance

-

display

of

parameter

-

press "enter"

key 3

times

-

display

of

response

time

-

enter

the new

response

time

störe

and

advance

to

next

parameter

-

display

of

parameter

-

switch

on

RATIO

-

display

of new

status

-

return

to

measuring

mode

09.83

3

OPERATION

- 19 -

S

Instruction manual

SFM

3.4.1

FIXED

WAVELENGTH

(continuation)

(with

calibration

and

blank)

0

l

[CALIBRATION

RUN J

[350 50.0

420 J

0

Blank

l

Sample

[AUTOBLANK

RUN J

[350

o.o

420"^)

l

o

put the

cell

containing

the

Standard sample into

the

cell

holder

-

press

key

"calib"

-

calibration

is

carried

out

-

display

in

measuring mode

put

the

cell containing

the

blank

solution into

the

cell holder

[350 48.4

420 J

-

press

key

"auto

blank"

-

Blank

value

is

measured

and

auto-

matically

put

into memory

-

display

in

measuring

modes

put

the

cell containing

the

sample

to be

measured into

the

cell

holder

-

result (relative

fluorescence)

03.84

3

OPERATION

- 20 -

K

Instruction manual

SPM

3.4.2 EMISSION

SCAN

(with

calibration

and

recording)

Example

with

following

parameters:

EXCITATION

EMISSION

SCAN RANGE

SCAN

SPEED

CALIBRATION

FACTOR

RESPONSE

360

nm

520 nm

(start

of

scan)

160 nm

80 nm / min

100

1.0

2.0

[

480

0.0

480J

l

2

enter

[EXCITATION

480

J

3

6

0

OFF

enter

EMISSION

480

]

5

2

0

^OFF

ente

EX.SCAN

ON

]

0

OFF

enter

EM.SCAN

OFF]

1

ON

enter

-

display

after

Initialisation

-

select parameter group

2

display

of

current excitation

wavelength

enter

new

value,

störe

and

advance

to

next parameter

display

of

current emission

wavelength

enter

new

value, störe

and

advance

to

next parameter

switch

off

EX-SCAN, störe

and

advane

to

next parameter

switch

on

EM-SCAN, störe

and

advance

to

next parameter

09.83

3

OPERATION

- 21 -

Instruction manual

SFM

3.4.2

EMISSION

SCAN

(continuation)

(with

calibration

and

recording)

[SCAN

RANGE

250 j

-

display

of

current

scan ränge

-

enter

new

scan ränge, störe

and

advance

to

next

parameter

SCAN

SPEED

100

]

display

of

current scan speed

enter

new

scan

speed,

störe

and

return

to

measuring

mode

(360

0.0 520 j

-

display

in

measuring

mode

3

enter

l

[CALIBRATION

100 J

-

Select parameter group

3

-

value

o.k.

HIGH

VOLTAGE

400 J

-

value o.k.

BLANK

0.0

]

-

value o.k.

FACTOR

1.0

]

-

value o.k.

[RESPONSE

i.o

j

-

display

of

current value

enter

new

value, störe

and

advance

to

next parameter

09.83

3

OPERATION

- 22 -

K

Instruction manual

SFM

3.4.2

EMISSION

SCAN

(continuation)

(with

calibration

and

recording)

[RATIO

OFF]

l

1

ON

return

[360

0.0 520 J

Sample

0

[360

5.0 520 J

( 360

OFLOW

500 J

CALIBRATION

RUN J

[360

100.0

402 J

-

display

of

current status

switch

on

ratio

and

return

to

measuring

mode

- put the

cell containing

the

sample

into

the

cell holder

-

fluorescence

of

sample

-

press

emission

key

until

the

fluorescence

is a

maximun

-

"0-FLOW"

appears

on the

display

-

press

"calib"

key

use

these keys

to

find

the

maximum

fluorescence

-

press

"calib11

key

-

calibration

is

carried

out

-

display after calibration

- set

recorder

parameters

(selectable

on the

recorder)

-

chart speed

- mV

ränge

(100mV)

-

zero

-

start position

-

press

"start/stop"

key

[3503.0

*520

l -

measuring

in

scan

mode

Emission

spectrum

is

drawn

on the

recorder

03.84

3

OPERATION

- 23 -

Instruction

manual

SFM

3.4.3

EXCITATION

SCAN

(with calibration

and

recording)

Example with

following

Parameters:

EXCITATION

EMISSION

SCAN

RANGE

SCAN

SPEED

CALIBRATION

FACTOR

RESPONSE

400

nm

(start

of

scan)

402 nm

100 nm

80 nm / min

100

1.0

2.0

[480

0.0 480

j

2

enter

[

EXCITATION

480

4

0

OFF

0

OFF

enter

[EMISSION

480J

4

0

OFF

2

enter

[EX.SCAN

OFF ]

-

display after

Initialisation

-

select

parameter

group

2

-

display

of

current excitation

wavelength

-

enter

new

value

display

of

current emission wave-

length

enter

new

value, störe

and

advance

to

next parameter

-

display

switch

on

EX-SCAN, störe

and

advance

to

next parameter

09.83

3

OPERATION

- 24 -

KK

Instruction

manual

SFM

3.4.3 EXCITATION SCAN

(continuation)

(with

calibration

and

recording)

[EM.

SCAN

ON]

0

OFF

enter

switch

off

EM-SCAN, störe

and

advance

to

next

parameter

[SCAN

RANGE

250]

l

1

ON

0

OFF

0

OFF

enter

[SCAN

SPEED

100

j

l

(400

0.0 402 J

-

display

of

current scan ränge

-

enter

new

scan ränge, störe

and

advance

to

next parameter

-

display

of

current scan speed

-

enter

new

value,

störe

and

return

to

measuring

mode

-

display

in

measuring

mode

3

enter

-

select parameter group

3

CALIBRATION

100 ]

-

display

of

current value

l

[HIGH

VOLTAGE

400

]

-

value o.k.

-

value o.k.

BLANK

0.0

]

-

value o.k.

[FACTOR

1.0

(RESPONSE

i.o]

2

enter

-

value o.k.

-

display

of

current value

-

enter

new

value, störe

and

advance

to

next parameter

09.83

3

OPERATION

- 25 -

Kl

Instruction

manual

SFM

3.4.3

EXCITATION

SCAN

(continuation)

(with

calibration

and

recording)

RATIO

OFF

[400

0.0 402

Sample

Q

-

display

of

current status

-

switch

on

ratio

and

retun

to

measuring

mode

- put the

cell

containing

the

sample

into

the

cell holder

[400

0.3

402 J

l

[400

OFLOW

402 J

[CALIBRATION

RUN

-

fluorescence

of

sample

-

press

"excitation"

key

until

fluorescence

is a

maximum

- if

"OFLOW" appears

on the

display

-

press

"calib"

key

use

these keys

to

find

the

maximum

fluorescence

-

press "calib"

key

-

calibration

is

carried

out

357

100.0

400 ]

[400

3.0 * 402 J

Excitations

spectrum

is

drawn

on the

recorder

-

display after calibration

- set

recorder parameter

(selectable

on the

recorder)

-

chart

speed

- mV

ränge

(100

mV)

-

zero

-

start position

-

press

key

"start/stop"

-

measuring

in

scan

mode

03.84

3

OPERATION

- 26 -

K

Instruction manual

SFM

3.4.4 SYNCHROSCAN
(with

calibration

and

recording)

Example

with

following

Parameters:

EXCITATION

EMISSION

SCAN RANGE

SCAN

SPEED

CALIBRATION

FACTOR

RESPONSE

440

nm

(start

of

scan)

500 nm

(start

pf

scan)

200 nm

80 nm / min

100

1.0

2.0

[480

0.0 480 J
l

2

enter

l

[EXCITATION

480

~)

4

4

0

OFF

[EMISSION

480 J

5

0

OFF

0

orr

enter

[EX.SCAN

OFF]

1

ON

enter

l

[EM.

SCAN

OFF")

-

display

after

initialisation

-

selection

of

parameter

group

2

display

of

current excitation

wavelength

enter

new

value,

störe

and

advance

to

next parameter

display

of

current

emission

wavelength

enter

new

value, störe

and

advance

to

next parameter

-

switch

on

EX-SCAN

-

switch

on

EM-SCAN

09.83

3

OPERATION

- 27 -

-S

Instruction manual

SFM

3.4.4 SYNCHROSCAN

(continuation)

(with

calibration

and

recording)

[

SCAN

RANGE

250]

-

display

of

current scan ränge

2

0

OFF

0

OFF

enter

l

[SCAN

SPEED

looj

-

enter

new

scan ränge, störe

and

advance

to

next parameter

-

display

of

current scan speed

8

—

0

OFF

return

[440

0.0

500J

l

l

[CALIBRATION

106"

enter

new

scan speed, störe

and

return

to

measuring

mode

-

display

in

measuring

mode

-

select parameter group

3

-

value

o.k.

l

[HIGH

VOLTAGE

400

-

value o.k.

j^BLANK

O.OJ

l

-

value o.k.

[

FACTOR

1.0

]

-

value o.k.

RESPONSE

1.0

J

-

display

of

current value

enter

new

value,

störe

and

advance

to

next parameter

09.83

3

OPERATION

- 28 -

Instruction

manual

SPM

3.4.4

SYNCHROSCAN

(continuation)

(with calibration

and

recording)

[RATIO

OFF

]

1

ON

return

[440

0.0

500~]

Sample

U[

[440

10.0

500 ]

Start

stop

[440 10.0 * 500 )

-

display

of

current

Status

—

switch

on

Ratio

and

return

in the

measuring

mode

- put the

cell

containing

the

sample

into

the

cell holder

-

fluorescence

of

sample

- set

recorder parameter:

(selectable

on the

recorder)

-

chart speed

- mV

ränge (1mV)

-

zero

-

start position

-

press

"start/stop"

Key

-

measuring

in

scan

mode

Synchroscan

spectrum

is

drawn

on the

recorder

The

A-EX

and

A-EM

of

maximum

fluorescence

is

measured

from

the

recorder diagram.

If

the

maximum

is too

small

or

outside

the

scale.

A

T

A

T

CALIBRATION

RUN

J

- use

EXITATION

und

EMISSION

to

set

required

wavelength

(A

Ex and

AEm)

-

press

"calib"

key

-

calibration

is

carried

out

340

100.00

400

i

start

stop

synchroscan spectrum

is

drawn

on

the

recorder

03.84

3

OPERATION

- 29 -

S

Instruction manual

SFM

3.5

CHANGING

THE

SPECTRAL

BAND

WIDTH

The

SFM 25 has

four selectable

slits.

The

Standard version

is

equipped

with 10/20

nm

emission-

and

10/15

nm

exitation slits. 5/10

nm

slits

are

also available

äs

accessories.

The

slit

selection

is

performed, without opening

the

monochromator,

from

the

underside

of the

Instrument.

Proceed

äs

described

below:

-

remove

all

plugs

at the

back

of the

Instrument

-

place a soft

cover

on the

right hand side

(foam

rubber, cardboard,

etc.)

- Tip up the

Instrument

over

the

right hand side base radius.

View

from below:

S1ÄNDWDSUTSEM.

:ICßOm)

EMSSON

(mortedwithgreen)

ENTRANCE

SUT

IN

: »nm

OUT : 20nm

STANDARD

SUTS

EX

:K)/15nm

IN : Wim

OUT : Gm

EMSSCN
EXITSUT

OPTIONAL

SLITS : S/IOtxn

(mofted

with

red)
IN : 5nm
OUT : lOrm

The

switching handles

on the

5/10

nm

slit

(Option)

are

marked

red and

on

the

10/20

nm

slit they

are

marked green.

09.84

3

OPERATION

- 30 -

Kü

Instruction

manual

SFM

3.6

SWITCHING

OFF THE

INSTRUMENT

The

SFM 25 can be

switched

off at the

mains

switch

at any

time.

AVOID

REPEATED SWITCHING

ON AND OFF IN

ORDER

TO

PREVENT

DAMAGE

TO THE

XENON

LAMP

AND

ELECTRONICS

09.83

3

OPERATION

- 31 -

S

Instruction

manua!

SFM

4.

ERROR

DETECTION

4.1

Error

messages

09.83

4

ERROR DETECTION

- l -

Instruction

manual

SFM

4.1

ERROR

MESSAGES

Operating

errors,

malfunctions

or

System errors

are

automatically

de-

tected

and

displayed.

To

continue

Operation again

after

an

operating error

or a

malfunc-

tion, press

the

"clear"

key.

For

additional

error

messages

see

chapter 6 (OPTIONS).

Error

table

1:

Error

message

0.

FLOW

U.

FLOW

SAMPLE COVER

OPN

ENTRY ERROR

WRONG

VALUE

PERIPH

NOT

READY

OPTION

NOT

INCL

HV-LIMIT

DO

CALIBRATION

BLANK

TOO

HIGH

LAMBDA

BUSY

Reason

emission

of

sample

too

high

emission

of

sample

too

small

sample

compartment

cover

open

in the

measuring

mode

wrong

key

pressed

Parameter

value

out of the

ränge

selected

pheriphery

not

connected

no

Option

The

automatic

cali—

bration

can not be

executed

within

the

available

high

vol-

tage

ränge . High

voltage

is too

large

or too

small

key

"auto

blank"

has

been

pressed before

a

calibration

has

been

carried

out

blank

value

for

"auto blank"

too

high

illegal input during
a

wavelength

posi-

tioning

Elimination

select

lower

high voltage

or

factor

or

calibrate again

select larger high voltage

or

fac-

tor, calibrate again

or

select

smaller blank value

close

sample compartment cover

delete

with "clear"

and

press correct

key

delete

value with "clear"

and

enter

new

value

delete

with

"clear"

check connections

and

peripheral

delete

with "clear"

delete

with "clear"

and

select

smaller

or

larger calibration

-

value,

or:

work without cali-

bration

delete

with

"clear"

and

calibrate

delete

with "clear

and

enter

required blank value

in the

para-

meter

mode

delete

with

"clear" . Start

input

after wavelength

has

positioning

has

finished

03.84

4

ERROR

DETECTIOM

-

2 -

H

Instruction manual

SFM

4.1

ERROR

MESSAGES

System

errors

are

recognised

during

the

self-check

and

displayed

afterwards.

If

such a message appears,

inform

your

KONTRON

Service

organization,

give

Information

about which error message

has

appeared

and

which Software Version your

Instrument

has.

The

Instrument

will

not

function.

Error

table

2:

Error

messages

ERROR

RAM X.

ERROR

EPROM

X.

ERROR

TRNS MOTCO

ERROR

TRNS MOTCO

ERROR

MOTOR

EX

ERROR

MOTOR

EM

ERROR

MOTCO

ERROR

TRNS ADDA

ERROR ADDA

EPROM

ERROR

ADDA

RAM

ERROR

ADDA

ERROR

LAMP DEFEC

ERROR

N.

FREQUENCY

Reason

error

in the

displayed

RAM

error

in the

displayed

EPROM

error

in the

data transfer

to

the

motor

control

error

in the

data transfer

from

the

motor control

error

in

monochromator

drive

excitation

error

in

monochromator

drive

emission

error

can not be

localized

on

motor

control

error

in the

data transfer

to

the

AD-DA Converter

error

in the

EPROM

of the

AD-DA

Converter

error

in RAM of the

AD-DA

converter

error

can not be

localized

on

AD-DA

converter

lamp

is

defect

or

badly

adjusted

mains

frequency outside

the

allowed

ränge

(45...

65

Hz)

09.83

4

ERROR

DETECTION

- 3 -

4

Instruction manual

SFM

6.

6.1

6.2

6.3

6.4

SPECIAL

ACCESSORIES

Thermostatted 4 cell changer

for

10x10

mm

Standard

cells

HPLC

flow

cell

with

holder

Slit

holder with

slit

5 and 10 nm

Slit

holder with slit

10 and 20 nm

91-00431

91-00430

91-00432

91-00481

6.5

Filter holder

for

excitation

and

emission

91-00434

6.6

Software extension pack

(Instr.

84-00095)

Plotter/Printer

and

method

files

91-00441

6.6.1 Storing a method

2

6.6.2

Recall

of a

method

file

3

6.6.3 Installation instruction

4

6.6.4

Operation

5

6.6.5

P800

used

äs

plotter

in

TIME DRIVE

6

6.6.6 P800

used

äs

plotter

in

LAMBDA DRIVE

8

6.6.7

How to

produce

an

overlay

on the

P800

10

6.7

Photomultiplier

R446

(200-800

nm)

91-00433

^

6.8

Polarization

- kit

(Instr.

84-00219)

91-00493

10.85

6

OPTIONS

- l -

18

Instruction manual

SFM

6.6

SOFTWARE EXTENSION PACK

91-00441

PRINTER/PLOTTER

- AND

METHOD

FILES

6.6.1

STORING A METHOD

(FILE)

Parameter

group

6 is

used

to

störe

the

method

parameters. A maximum

of

9

methods

can be

stored.

Example:

[360

fff.f

480

J

6

enter

[COPY

TO

FILE

1.

-

select

parameter

group

6

4

—

enter

1

or

return

[360

fff.f

480 J

the

parameters

are

copied from

the

working

file

into

one of the

method

files

(file

1.4)

and the

Instrument

returns

to the

measuring

mode.

09.83

6

OPTIONS

—

2 —

Kü

Instruction

manual

SFM

6.6.2 RECALL

OF A

METHOD

FILE

Stored methods

can be

recalled

at any

time

(providing

the

Instrument

is

not in a

scan

mode).

The

method parameters

are

copied form

the

mehtod

file

into

the

working

file

(the data also

remains

in the

method

file).

The

files

can be

recalled using

the

direct

or

indirect selection:

Examples:

a)

direct

selection

[480

0.0

480J

-

display after

Initialisation

recall file

1.4 and

return

to

measuring

mode

350

fff.f

480 J

Parameters

for

method

1.4 are

set up

b)

indirect

selection

'480

0.0

480 J

1

[FILE

1.1

OFF]

l

enter—enter

[FILE

1.4

OFF)

1

ON

return

[350

fff.f

480

J

-

display after

Initialisation

-

select

parameter

group

l

press "enter"

until

the

required

file

is

displayed

recall file

1.4 and

return

to

measuring

mode

parameter

for

method

1.4 are set

up

09.83

6

OPTIONS

- 3 -

Instruction

manual

SFM

6.6.3

INSTALLATION

INSTRUCTIONS

This

extension pack

allows

communication

between

SFM 25 and

KONTRON'S

P800,

supposed

the

P800

is

supplied

with

an

RS-232C

Interface.

The SFM

25

programs

the

P800

via the

parameter group

4.

This

instruction

only

explains

the

Operation

of

P800

via the

keyboard

of

SFM

25.

A

detailed instruction manual

is

included with each

printer/plotter.

Gable

Connections

SFM

25

Location

of

MEMORY

BOARD

94-90750

P

800

\

T

T

T

-i

_

0

,

no

r\r\ A c:

i

III

1

°

^

e

1

*

13/

-*-

- •

i

1

.

1

WARNIHQ

^^^fjjSKSq

e

(^)

e

Replacing

the

Software pack

- On the

back

of the

Instrument,

remove

cover sheets

l an 2

-

Pull

out the

MEMORY

BOARD

94-00750

-

Remove

the

MEMORY

PACK

by

pressing

the 2

snappers

while

pulling

the

MEMORY

PACK

off.

- On the

new

Memory Board

94-90750

switch

Sl

to ON, to

allow

saving

of

files.

- Fit in the new

MEMORY

EXT.PACK

and

snap

it on

tightly.

-

Switch

on Sl

-

Plug back

the

MEMORY

BOARD

and

mount

the

cover sheets.

Pin

assignement

on

cable

93-00545

Printer-P800

(plug)

SFM 25

(socket)

13

DTR

DSR

/ 6

F-GND/1

\

•

^

*^

25

25

20

orongc

20

red

\

\n

~\k

r^O

n

0

o

o

o

o

o

0

0

0

°

O

o

0

°

^0

•Q

5/

CTS

2l

TXD

1/F-GNO

shietd

10.85

6

OPTIONS

- 4 -

Instruction manual

SFM

6.6.3

INSTALLATION

INSTRUCTIONS

(cont.)

Installation

-

Connect

Instruments

according

to

fig.

-

Prepare

the

P800

according

to the

instructions

in the

manual

of

P800.

-

Switch

on

Instruments.

-

After

the

SELF

CHECK

of SFM 25 is

finished

the

BAUD

RATE

in

Parameter group 5 (INTERFACE CONTROL)

must

be

adjusted

to the

baud

rate

of

P800.

The

preset

value

of

BSÜB

BAUD

RATE

is

4800.

It

is

advisable

to set

P800

to the

same rate,

in

order

to

eliminate

adjusting

on

each

start/up.

6.6.4

OPERATION

^

Following

operating

modes

are

programmable:

Standard Software

U-Fix)

U-Scan)

Software Extension Pack

P800

used

äs

printer

If

P800

is

employed

äs a printer, then follow

the

description

in the

SFM

25

INSTRUCTION

MANUAL

chapter

3,

pages

15, 16 and 17.

02.84

6

OPTIONS

-

5 -

Instruction

manual

SFM

6.6.5 P800

used

äs

plotter

in

TIME DRIVE

In

this operating mode

the

fluorescence

value

is

plotted against time.

The

parameters

BJM

EX

SCAN

and

BälHI

EM

SCAN

are set to

"off".

With

the

"start/stop"

key the

TIME DRIVE

can be

started

and

finished.

Example

for

TIME

DRIVE:

[342

100.0

482~]

l

[

TIME DRIVE

OFF~)

l

TIME SCALE

(342

100.0

482

]

_L

342 T

100.0 T 482

J

[PLOT

AXIS

RUN]

PLOT AXIS BRAKE

[ 342

100.0

482J

Calibration

to

100.0

The

Instrument

is

prepared

for

plotting

on the

P800

Range:

0.01

....

20

min/cm

Start

plotting

The

display indicates

the

opera-

ting mode

T.

The

functions "PARAMETER

VIEWING"

chapter

3,

page

14 and

"READ"

chapter

3,

page

7 are

accessable.

However,

parameters

can not be

changed.

End

of

measurement

P800

plots

and

labels

the

axes

and

prints

out the

table

of

important

parameters.

Labelling

can be

abor-

ted

by

pressing

the

"return"

key

02.84

6

OPTIONS

-

6 -

3

Instruction manual

SFM

6.6.5

P800

used

äs

plotter

in

TIME

DRIVE

(cont.)

129*

100*

SFM

SAMPLEI
CODE
TIME

DRIVE

TIME

SCALE
EXCITATION
EMISSION
CALIBRATION
HIGH

VOLTAQE

BLANK

FACTOR
RESPONSE

-

2 5

18-01-1984

20.

13

min

2.88 m intern

342 nm
482 nm

100.0

457

V

0.0

1.00

8.8

sac

80s;-

-

60*--

40*--

20*--

8.00

22.00

min

Remarks:

In

TIME DRIVE each printout

is

limited

to

1900

mm

length.

If

the

printout

exceeds this

limit,

the

display

of the SFM 25

shows

"TIME

AXIS

O.FLOW".

After pressing

the

"clear"

key the

plotting

is

completed

with

the

description.

The

exact

running

time

in

TIME DRIVE

is

printed

out in the

parameter

table.

e.g.

TIME DRIVE 20.13

min

02.84

6

OPTIONS

-

7 -

H

Instruction

manual

SFM

6.6.6

P800

used

äs

plotter

in

LAMBDA

DRIVE

In the

LAMBDA

DRIVE

mode

the

fluorescence

value

versus

wavelength

is

plotted.

This

is

done

in

Ascan

Operation

only.

Therefore a .A-scan

mode

must

be

programmed

in

parameter group

2,

WAVELENGTH

CONTROL.

Example

for

plotting

the

emission

spectrum.

[342

100.0

482 J

Calibration

at

342/482

to

100.0

4

-

•

8

enter

[LAMBDA

DRIVE

OFF J

l

1

ON

enter

[LAMBDA

SCALE

20 J

The

Instrument

is

prepared

for

plotting

on the

P800

Range:

l ... 100

nm/cm

[342

100.0

482 J

start
stop

1

342

10.4

* 502 ]

PLOT

AXIS

RUN

The

Instrument

runs

in

scan

mode.

The

P800

plots

the

fluorescence
versus emission wavelength. Scan­ning

can be

interrupted

and

restarted

with

the

"start/stop"

key. After completion

or

abortion

of the

plot

the

P800

plots

the

axes

and

labels

them.

Plotting

the

axes

can be

aborted

by

pressing

the

"return"

key.

02.84

6

OPTIONS

-

8 -

hK

Instruction

manual

SFM

6.6.6

P800

used

äs

plotter

in

LAMBDA

DRIVE

(cont.)

SFM

CODE

SCAN

LAMBDA

SCALE

SCAN

SPEED
EXCITATIOH
EMISSION

CALIBRATION
HISH

VOLT

ÄSE
BLANK
FACTOR

RESPONSE

- 2 5

1S-91-19S4
EMISSION
20

nrn/crn

180

nmxTni

n

342

nm

600

nm

180.0

540

V

0.0

1.00

2.3

se

c

500

600

nrn

^

Since

the

recording

speed

and the

paper

length

for

each recording

is

limited

the

parameters

SCAN

SPEED,

SCAN RANGE,

and

LAMBDA SCALE

are

monitored. LAMBDA SCALE

is

automatically

adjusted,

if the

following

ratlos

exceed

the

limits.

SCAN

RANGE

LAMBDA

SCALE

=

Paper length

(max.

1570

mm)

and

SCAN

SPEED

LAMBDA

SCALE

=

Recording speed (see

table

chapter

2,

page

15)

02.84

6

OPTIONS

- 9 -

-K

Instruction manual

SFM

6.6.7

How to

produce

an

overlay

on the

P800

Mark

the

starting position

of the

first

plot,

then

run the

first

scan.

With

the

paper advance button

move

the

paper back

to the

starting

mark.

If

desired

set new

parameters

and

start

the

overlay plot.

This

procedure

can be

repeated

any

number

of

times.

Table

for

scales

and

labelling

of

X-axis

gJCT

LAMBDA

SCALE

Scale

(nm/cm)

1

2 - 3

4 - 7

8-15

16

- 30

31

- 70

70

- 100

short

marks

at:

increment

(nm)

1

2

5

10

25

50

50

long

marks

at:

increment

(nm)

5

10 .

10

20

50

100

100

label

at:

increment ( nm

)

5

10

10

20

50

100

200

02.84

6

OPTIONS

- 10 -

3

Instruction manual

SFM

6.6.8

Automatic

curve

overlay

with

P80O

Before

starting

an

automatic

overlay

the

parameter

values

in

group

2

must

be set

accordingly.

WAVELENGTH

CONTROL

S.IEXOTATION l («m)

200 800 .

J.2EM1SSK3« « ((im)

200

800

480

UEXCfTAflOMSCAN

O«/OFF

OFF

2.4

EMISSION

SCAN

OK

(OFF

OFF

2.SSCANRANGC«

'

3.6SCflHSPEEDWm/<nml|

20 500 l 100

2JOVERIAY

l

OH/OFF!

OFF

DATA CONTROL

UHKiHVOtTACEtCVEl

«XI

999

OH/OFF l OFF

PRINTER

/PLOTTER

INTERFACE CONTROL

SAVERLE

Example

Sample:

Quinine

sulfate

Solution:

0.1 n

Sulfuric

acid

(350

100 450 ]

{OVERLAY

OFF

J—

-

Calibration

with

sample

at

maximum

emission.

Activation

of the

automatic

"Overlay"

[350

100

450

-

Return

to

measuring

mode

05.86

6

OPTIONS

- 11 -

IS

Instruction

manuai

SFM

6.6.8

Automatic

curve

overlay

with

P800

(cont.)

LAMBDA

DRIVE MODE

—'

retum

350

ff.f

0 550

[350

ff.f Q 550

]

[350 0 ff.f

0

37ÖJ

350 0 ff.f 0 550]

Activation

of

plotter

in

lambda-

drive

mode

-

Start

wavelength

The

"0" in the

right

display

field

(scan

digit) indicates

the

programmed

automatic

overlay.

-

Start

of

reference recording

-

Measurement

End

of

measurement

The

"0" in the

left

display field

(stop

digit) indicates

that

an

overlay recording

is

following.

-

Start wavelength

All

following measurements will

be

recorded

in

overlay mode.

Note:

If

the

scan ränge

is

shortened,

then

the new

values will define

the

curve

length.

If the

scan ränge

is

selected longer, then

the

curve length remains

the

same

and the

leftover will

be

ignored.

By

pressing

the

"clear"-key

a new

reference curve

can be

recorded.

05.86

6

OPTIONS

- 12 -

13

Instruction

manual

SFM

6.6.9

BASELINE

CORRECTION

The

SFM 25 has

been designed

äs a single

beam

Instrument

(for

energy

saving).

Therefore,

it

won't

compensate

the

strongly wavelength

depending

exitation energy

.

The

baseline correction,

however,

will compensate

for the

wavelength

dependency

and

give a true

measuring

value.

Measuring

the

baseline

Let

the

Instrument

warm

up to

operating temperature (appr.

/£h)

if

possible

with cuvette holder

and the

cuvette filled with solution.

Note:

Calibrate

the

Instrument

and be

sure

no

measurement

values

are

in

the

"0-FL"

over

flow

or

"U-FL"

under

flow

ränge.

Before storing a baseline

the

parameters

in

group 2 must

be set

accordingly.

If an

Overlay

is

desired

it

must

be

programmed

(see

6.6.8).

The

"scan

speed"

should

be

selected

below

180

nm/min

otherwise

Interpolation

errors

may

occur.

Example

Sample:

Quinine

sulfate

Solution:

0.1 n

Sulfuric acid

(350

100 450 ]

[BASELINE

SAVE

OFFJ-

350

ff.f

550 ]

-

Calibration with

sample

at

maximum

emission

-

Activation

of

Baseline

storage

-

Start

wavelength

05.86

6

OPTIONS

- 13 -

Kl

Instruction

manual

SFM

6.6.9

BASELINE

CORRECTION

(cont.)

[LAMBDA

DRIVE

OFF

1

ON

return

[350

ff.f

550 )

[350

ff.f

* 550 ]

[350 ff.f

370 ]

Instruction

set

only

needed

if

recording

of the

baseline

is

desired

on the

plotter.

Solution

is in the

light

beam.

Wait

until

fluorescence

value

is

stable.

-

Baseline

start

-

Measurement

- End of

measurement

The

baseline measurement will

be

saved under

7.2 and

with

"BASELINE

SAVED"

acknowledged.

05.86

6

OPTIONS

- 14 -

KS

Instruction

manual

SFM

6.6.9 BASELINE

CORRECTION

(cont.)

Recording

of a

non-corrected

emission spectrum

with

sample

in the

light

beam.

[BASELINE

SAVE

ON

J—

[350 ff.f

550 ]

4

—

•

—

8

[LAMBDA

DRIVE OFF"

350

ff.f

550

[350

ff.f

* 550 ]

[350 ff.f

370 ]

-

Baseline

off

Instruction

set

only

needed

if a

recording

of the

none corrected

emission spectrum

is

desired

on the

plotter.

-

Start wavelength

for

sample

-

Lambda

scan

start

-

Measurement

- End of

measurement

05.86

6

OPTIONS

- 15 -

Instruction manual

SFM

6.6.9 BASELINE

CORRECTION

(cont.)

After

the

baseline measurement

all

following measurements

in the

corresponding

lambda

scan

mode

can be

corrected

with

the

baseline.

Do

not

change

the

parameters

2.1...2.4

.

Recording

of the

corrected emission spectrum.

[SSO

ff.f

550

]

'BASELINIE

SAVED

]-

[BASELINE

CORR

OFF]

350ff.f

550J

[350 ff.f

* 550 ]

[350 ff.f

370 ]

—

Start wavelength

for

sample

- The

baseline

is

saved

—

Activation

of

baseline correction

-

Start

of

measurement

-

Measurement

- End of

measurement

05.86

6

OPTIONS

- 16 -

BÖ

Instruction

manual

SFM

6.6.9 BASELINE

CORRECTION

(cont.)

The

following

graph shows

an

overlay

recording

of the

three

emission

spectrums.

120*

108*--

^

S F M - 2 5

SAMPLE!
CODE
SCAN
LAMBDA

SCALE

SCAN

SPEED
EXCITATION
EMISSION

CALIBRATION
HIGH

VOLTAGE
BLANK
FACTOR
RESPONSE
BASELINE

SAVE

30-04-1336
EMISSION
20

nrn/cm

100

nmxmi

n

350

nrn

550

nm

100.0

737

V

0. 0

l

.00

S.0

sec

O

N

88*--

60*

- -

40*

20*--

0*

Sample

not

corrected

Sample

corrected

370

406

450

500

556 n ru

05.86

6

OPTIONS

- 17 -

4

Instruction manual

SFM

7.

APPENDIX

7.1

Optical diagram

7.2

Mechanical

construction

7.3

Electronic block diagram

7.4

Software block

7.5

Sample

cells

7.6

Function check

7.7

Communication

via RS 232 11

7.7.1 Format

of

data-out-string

11

7.7.2

Code

table - Interface

RS 232 12

7.7.3

SFM 25

entry mask

13

7.7.4 Programming Examples

14

11.84

7

APPENDIX

- l -

Instruction

manual

SFM

7.1

Optical

diagram

Description:

1)

Xenon high pressure

lamp

2)

Colimating

lens

3)

Selectable

slit

at

the

entrance

to the

excitation

monochromator

4)

Beam

splitter

5)

Reference detector (photo diode)

6)

Excitation

grating

7)

Selectable slit

at the

exit

of the

excitation monochromator

8)

Chopper

9)

Plane

mirror

10)

Colimating lens

11)

Sample

in

cell

12)

Colimating lens

13)

Selectable slit

at the

entrance

to the

emission

monochromator

14)

Emission grating

15)

Selectable slit

at the

exit

of the

emissions

monochromator

16)

Photomultiplier

09.83

7

APPENDIX

- 2 -

-S

Instruction manual

SFM

7.2

MECHANICAL

CONSTRUCTTON

^

Ignition

unit

X"T\7

v

\

\

P

ü

n

ii

!.

'l

|!

Power

supply',1

II

1

1 1

1

II

1

j

II

i

Ex-

monochromator

Sample
compartment

Em-

monochromator

Housing

D D

Electronic

Mains

switch

10.84

7

APPENDIX

-

3 -

9

Instruction

manual

SFM

7.3

ELECTRONIC BLOCK

DIAGRAM

/^ N Lamp

power

(

)

—

supply

and

^ — '

ignition

,

Motor

/

O

mi-it-rol

\"

,

,.

_

_..,

@

l

.

A

AD-DA

/*

©

Converter

\

_^

,.

yvy

\

^^f^

Analog

Outputs

for

relative

fluorescence

and

wavelength

Power
Supply

K

\d

+

/

Display

board

/

\

K

\

R^s

^^5*»

/

Processor

ino-^oz.)

N

V

1

r^

P

Remote

K

/

Memory

>

\e

;

>

Accessable

x

from

outside

board

.

09.83

7

APPENDIX

- 4 -

'Kl

Instruction

manual

SFM

7.4

SOFTWAREBLOCK

S E L

GT^^-^

Parameter
entry

retum

F C H E C K

Main

progi

(ir^-^

-_

Measurement

in

Fix

mode

Through

selec-

tion

of

another

mode

r

am

735

I±±-^

Measurement

in

Scan

mode

Automatic

at the

end

of a

scan

or

via

keyboard

start

t

stop

"öT^^-—

_

Error message

cle&r

09.83

7

APPENDIX

- 5 -

Instruction

manual

SFM

7.5

SAMPLE

CELLS

The

SFM 25 is

fitted with a constant temperature

cell

holders

for

Stan-

dard

rectangular

10 x 10 mm

cells.

Modern cells

are

normally constructed from pieces

of

glass

or

silica

which

are

fused together

to

form a liquid-tight

Container (cells used

for

fluorescence must

not be

cemented because

the

glue

or

cement

in

many

cases

shows

fluorescence).

Glass

is

suitable

for the

ränge

of 340 -

1000

nm

but

silica

is

necessary

for

work

between

220 and 340 nm.

Cells

for

fluorescence have three

or

four

optical

faces.

Standard cells

are

usually supplied with a simple glass

or

plastic

lid

to

prevent spillage

when

lifting

them

in or out of the

sample compart-

ment.

If

volatile solvents

are to be

used

it is

desirable

to use

cells

with

PTFE stoppers

to

avoid changes

of

fluorescence

due to

evaporation

and

to

prevent

the

escape

of

vapours into

the

Instrument.

When using

stoppered

cells care should

be

taken

to

leave

an air gap at the top or

the

cell

may be

ruptured

by any

build-up

in

pressure.

Cell

Cleaning

As

fluorometric analysis

is a

high sensitivity

measuring

technique,

considerable care

in

cleaning

and

storage

of

sample cells (pipettes

and

glassware)

is

required. Glass pipettes

and

other glassware should

be

soaked

in

chrpmic acid overnight, rinsed

at

least

five

times

with

tap

water

and

soaked

in

distilled

or

deionized water

for

about

10

minutes.

Then rinse again with distilled

or

deionized water.

Allow

to

drain

and

air

dry.

For

sample cells, a special cleaning procedure

is

required.

The

easiest

way

to

keep sample cells

clean,

is

rinsing

them

always

immediately

after

use

with distilled

or

deionised water.

It

is not

recommended

to

soak cells

for a

long

period

in

chromic acid.

Special cell

cleaning

Solutions

are

preferred.

Use

these

Solutions

according

to the

manufacturer's

instructions.

After carefully rinsing

the

cells with distilled

or

deionized water,

allow

to

drain

and air

dry.

Never

dry

cells

in a

heater.

Never

wipe sample cells with

the

laboratory

coat.

Use

only

soft

paper tissues (not

the

special white ones,

they

are

threated

with optical elucidaters which

show

bright

fluorescence)

or

special optical

lens

tissues

such

äs

"Kimwipes".

04.84

7

APPENDIX

- 6 -

Kü

Instruction

manual

SFM

7.6

FUNCTION

CHECK

The

"operating

summary"

will

help

you to

perform

the

function check.

-

Prepare

the

Instrument

for

Operation,

according

to the

Installation

instructions.

Set all

slits

to 10 nm.

-

Plug

in the

mains

connection

and

switch

on the

mains switch

The

lamp

should ignite after

l to 3

ignitions (audible

noise).

The

radiation

can be

seen

with

help

of a

white card

(see 1.5.4).

For 3 min.

'SELF

CHECK1

will

be

shown

on the

display. After this

time

the

Instrument

switches

automatically into

the A

-fix

measuring

mode.

-

Close

the

sample

compartment

[480

0.0

480 J

appears

on the

display

-

Open sample compartment

[SAMPLE

COVER

OPENJ

appears

on the

display

Install

cell

holder

and put

the

cell filled with

bidistilled

water

into

the

holder

-

Close sample

compartment

cover

and

press

key

[

CALIBRATICN

RUNJ

appears

and

changes

after

few

seconds

to

[480

100.0

480

J

The

displayed

value

should

be 100 +

1.0.

-

Check

the

high voltage value

by

pressing

keys:

The

displayed value shculd

be

<

600 V.

07.86

7

APPENDIX

-

7 -

IS

Instruction manual

SFM

7.6

FUNCTION CHECK

(continuation)

The

following

parameter

values should

be set up

under "Data

Control":

jSIHi

CALIBRATION 100.0

jcWai

HIGH

VOLTAGE

(do not

change)

BLANK

FACTOR

RESPONSE

RATIO

SELFCHECK

10.0

0.1

l

OFF

OFF

Return

to the

A-fix-mode

with

key

[ 480

9.0

480J

This

value

should

be

exact

9+1

-

Following

parameters

must

be

reset:

EE1

BLANK

90.0

FACTOR

10.0

RATIO

ON

^80

100.0

480J

This

value should

be 100

+10.0

X - Press

-Search-key

(excitation)

The

value

of the

excitation

wavelength

increases

slowly.

approx.

4 sec the

speed increases.

After

A

- Press

-Search-key

(excitation)

The

value

of the

excitation wavelength decreases slowly.

approx. 4 sec.

the

speed

increases.

After

- The

same

test

can

be

performed

with

the

A-Search-keys

of

emission

09.83

7

APPENDIX

-

8 -

K

Instruction manual

SFM

7.6

FÜNCTION CHECK

(continuation)

Function

check with

Printer

and

Recorder

-

connect printer

and

recorder (X,Y)

-

Switch

on

printer

and

recorder

-

Wait

until

Initialisation

procedure

is

finished (about 3 min.)

[ 480 0.0 480 ]

-

Press

key

The

values

on the

display will

be

printed out.

-

Select

"LIST

PARAMETER" with

the key

sequence:

CODE enter a 10-digit

numerical code

PARAMETERLIST "ON"

All

parameters

of

"WAVELENGTH

CONTROL"

and

"DATA

CONTROL"

are

printed

out.

SftMPLE:

CODE

13. 07. E3

WBVELENGTH

CDNTROL

EXCITBTIDN

EMISSION

EX.

SCAN

EM.

SCAN

SCON

RftNGE

SCBN

SPEED

DflTft

CONTRDL

CflLI

ERBT

ION

HIGH

VOLTflGE

FOCTOR

BLANK

RESPONSE

RflTIO

350

nw

337

nm

OFF
OFF

25B

um

100

nm/min

100.0

502

1.B0

2.0

4.

0 sec

ON

03.84

7

APPENDIX

-

9 -

K

Instruction manual

SFM

7.6

FUNCTION

CHECK

(continuation)

Program

for

X-Scan:

•3CT

EXCITATION

350

PJU

EMISSION

397

•aCT

EXCITATION

SCAN

OFF

•aCT

EMISSION

SCAN

ON

•aCl

SCAN RANGE

100

•ata

SCAN

SPEED

100

-

Return

to the

measuring

mode

with "return"

key.

Place a cell

containing bidistilled water

in the

cell holder

and

close

the

sample

compartment

cover.

-

Calibrate

the

Instrument

with

key

Set

the

emission wavelength

to 470

nm

[

EMISSION

470

]

-

Return

to the

measuring

mode

.with

the

"return"

key

Prepare

recorder

Y-input:

ränge

= 100 mV for

füll

scale

X-input: ränge

= 100 mV for

about

30 cm

-

Press

Scan

"Start"-key

The

recorder shows

the

Raman

band

of the

water

at an

excitation

of 350

nm.

09.83

7

APPENDIX

- 10 -

Instruction

manual

SFM

7.7

COMMUNICATION

VIA RS 232

7.7.1

FORMAT

OF

DATA-OÜT-STRING

X X

X

SP

Y

Y

Y

SP

V_

F

F

F

•

F

SP

S1

S2

S3

CR

LF

A-EX

DATA

A-EM DATA FLUORESCENCE INTENSITY STATUS

The

letter

"0"

must

be

sent

in

order

to

switch

ON-LINE

CONTROL

to on or

off.

With

"ON", a complete

string

is

Output.

With

"OFF",

a

string

without

status will

be

output.

Status

Byte (Sl)

ERROR

and

MODES

ERRORS

A

7

6

5

4

3

2

1

0

MOD]

•

uuvc-n

—

^Qo-h

-Ho

"1

l'

MODES

1-8

(see

table

7.7.3)

-

ERROR MESSAGE

ON

DISPLAY (press CLEAR key)

OPEN

Status

Byte

(S2)

Working state

of SFM 25

7

6

5

4

3

2

1

0

•.

M

-LAMBDA

BUSY

(Positioning)

ADDA

BUSY

(e.g.

CALIB

RUN)

—MEAN

RESULT

(valid)

~MEAN

BUSY

ON

DISPLAY

Set to "1"

Status Byte (S3)

State

of

accessories

0

STATUS

OF

POL.FILTER

(REMOTE

"l"/MAN

"0")

—

BASELINE SAVED

BASELINE

CORRECTION "RUN"

'-LOCK

(ON)

to "l

10.86

7

APPENDIX

- 11 -

Instruction

manual

SFM

7.7.2 CODE

TABLE - INTERFACE

RS 232

ASCII

0

1

2
3
4

5

6
7

8

9

•

_

C

E

R

S

P

B

Z

A

D

L
0

X

T

M

N
U

V

(DEL)

(CR)

HEX

30
31
32
33
34
35
36
37
38
39
2E

2D

43

45

52

53
50
42
5A
41

44
4C
4F

58
54

4D

4E

55

56

7F

ÖD

FUNCTION

0
1

2
3
4

5
6
7
8

9

•

_

CLEAR
ENTER

RETURN

START/STOP

PRINT/READ

AUTO

BLANK

AUTO

ZERO

(f)

CALIBRATE

DATA

OUT

KEYBOARD

LOCK

ON

LINE CONTROL

ON

LINE CONTROL
DISPLAY TEXT
POLAR.

FILTER

H

POLAR. FILTER

V

ANAL.

FILTER

H

ANAL.

FILTER

V

DELETE

CAR.

RETURN

DESCRIPTION

"S

>

Numeric

entry

s

For

code (4.1)

Error

acceptance

>

Ciosing

instruction

Lambda

scan (time-drive)

To

generate average

value

To

adjust blank + factor

To

adjust

blank

(factor

remains)

To

adjust

HV

T3

03
O

0)

•*•

JC

-p

•H
3

M

T3

C
O

a

CD

0)

0

o

Transmit

data—

string

once
Keyboard disable
To

insert

datastring

status

(ON)

To

insert

datastring

status

(OFF)

Text

in

ASCII,

max.

length

62

signs

1

?

Control

of

polarization accessory

J

Delete last sign

in

instruction

Terminate

instruction

string

The

definition

of the

functions

remain

the

same,

whether

SFM 25 is

keyboard

or

Interface

operated.

10.84

7

APPENDIX

- 12 -

Instruction

manual

SFM

7.7.3

SFM 25

ENTRY

MASK

~~~~""---^^^

MODE

and

FUNCTION~~^—

-^NUMBER
^~^-\

NUMMER/0-9

+/-

.

(DP)

CALIBRATE
AUTOBLANK

CLEAR
ENTER
RETURN

START/STOP
PRINT/READ

(MEAN)

NOT

USED

(f)

SEARCH

(Keyboard

only)

DATA-OUT
KEYBOARD-LOCK
ON/OFF
ON-LINE-CONT.

ON

ON-LINE-CONT.

OFF

DISPLAY-TEXT

POLAR

FILTER

H

POLAR

FILTER

V

ANAL

FILTER

H

ANAL

FILTER

V

DELETE
CLOSING COMMAND

for

RS 232

ASCII

0-9

-

•

A

B

C

E

R

S
P
F

D
L

0

X

T

M

N

U

V

(7FH)
(ODH)

INIT

(D

X

X
X
X
X

X
X

X

-

FIX

(2)

X

X
X

X

X

X

X

X
X

X

X

X

X
X

X
X

X
X

PAR/

ENTER

(3)

X
X

X

X

X
X

X

X

X

X
X
X

X

X
X

Ä-

SCAN

(4)

X

X

X

X
X

X
X
X
X

X
X

STOP

(5)

X

X

X

X

X

X

X

X
X
X

X

X
X

SEARCH

(6)

X

X

X

X

X
X
X

X

X
X

ERR

(7)

X

X

X
X

X

X

X

TIME

DRIVE

(8)

X

X

X
X

X

X

X

X
X

X
X

X

X

VIEW

X

X

X

X

X
X
X
X

X
X

"C" is

also used

to

clear

resetable ERRORS (see

Error

tables

4.1]

"R"

abortes describing

of the

axes

on

Plotter

The

Interface

has no

access

to the

"SEARCH"

keys

10.84

7

APPENDIX

- 13 -

tut

Instruction manual

SFM

7=7.4

PROGRAMMING

EXAMPLES

Programming example

for

data interchange

between

PC-8201A

and SFM 25

in

BASIC.

l.

Instrument control

The

following program

allows

to

operate

the SFM 25 via

the~

Computer

terminal.

'!

-7l

TM

C;

23

OFEN

"COM37N8iXS"

FOR

OU^FU7*

H3

Si

33

IMPUT

"IM3TRUCTIONB"ia$

43

PRINTM,Fi*

50

GOTO

30

53

GLDSE

Line

20

initializes

the RS 232

Interface.

The

interface

parameters

from

SFM

25 and

PC-8201

must

match.

(Both

are

programmable)

Operation: - Start

program

REM.BA

-

Input

instruction

string

according

to

code

table

-

SFM

25

executes

the

instructions

2.

Reception

of

data

strings

from

SFM

25

The

following program

allows

to

receive a data

string

from

SFM

25.

:-[7)

npphj

"CGM " 7WS1

V.

"•'

FO«

T;MP;

'3

INPUT#1,B*

Operation: - Start

program

REC.BA

- On

SFM

25

press

key

"print/read"

-

After completing

the

Integration

time,

the

entire

data

string

is

shown

on the

display.

11.84

7

APPENDIX

- 14 -

19

Instruction manual

SFM

^

_____

7.7.4

PROGRAMMING

EXAMPLES

(cont.)

3.

Control

program

With

simple

combinations

of the

two

programs,

you can

realize

complete

communication

programs.

20

QPH!N

HCGn;

7NS1XB"

FOR

OUTPUT

PiS

*i

33

OPEN

"COlsTNSlXS"

FOR

INPUT

HS

*2

>:J£

TNiOyT " T

MpPiJPTT

niMQ " ;

Ci*

53

PRIMTtfl^

M$

g g T M p!

:Ti|:2-

p*.

70

Si=flSC(MID'*(B$i

IG?

1)

)

30

52=RRC'!y!r

D* f PS---

17i 1 ) ')

30

S3=HSC(M!D*CB3>;

iR, i )

)

l 30

PR

INT N T

DS

i"Rs7

': , i

/;.

"i

:

!i

5 S *

~: " ' R l - "

S^' — " * 5'^' * "

-~r—."

113

LPRIMT

MID*CB*t

i*

14)

?"

:Sl =

";Si5"

B2="IS25:1

S3=="

123

GOTD

40

130

CLOSE

Operation: - Start

program

-

Input

instructions

on PC

terminal

-

Terminate

the

instruction string

with

"D" or "P" in

order

to get a

data

string

from

SFM

25.

-

Measuring

data

and

decoded

Status

words

will

be

Output

to

display

and

printer.

Examples

for

entries:

^

Example

for

measuring

data

Outputs

to

display

and

printer

11.84

7

APPENDIX

- 15 -

a

Instruction

manual

SFM

7.7.4

PROGRAMMING

EXAMPLES

Format

of

data string

'«'™Y

l

Meaning

Wavelength

Wavelength

Wavelength

Ist

numeral

2nd

numeral

3rd

numeral

Em.

wavelength

Ist

numeral

Em.

wavelength

2nd

numeral

Em.

wavelength

3rd

numeral

sign

Fluorescence

Ist

numeral

Fluorescence

2nd

numeral

Fluorescence

3rd

numeral

decimal

point

Fluorescence

4th

numeral

Ist

Status word

2nd

Status word

3rd

status word

Termination carrage return

of

instruction l ine

feed

11.84

7

APPENDIX

- 16 -