MOLECULAR SPECTROSCOPY
RamanMicro 200 Series
Getting Started Guide
Release History
Part Number Release Publication Date
L1321911 B February 2011
Any comments about the documentation for this product should be addressed to:
User Assistance
PerkinElmer
Chalfont Road
Seer Green
Beaconsfield
Buckinghamshire
HP9 2FX
United Kingdom
Or emailed to: info@perkinelmer.com
Notices
The information contained in this document is subject to change without notice.
Except as specifically set forth in its terms and conditions of sale, PerkinElmer makes no
warranty of any kind with regard to this document, including, but not limited to, the
implied warranties of merchantability and fitness for a particular purpose.
PerkinElmer shall not be liable for errors contained herein for incidental consequential damages in
connection with furnishing, performance or use of this material.
Copyright Information
This document contains proprietary information that is protected by copyright.
All rights are reserved. No part of this publication may be reproduced in any form whatsoever or
translated into any language without the prior, written permission of PerkinElmer, Inc.
Copyright © 2011 PerkinElmer, Inc.
Produced in the U.K.
Trademarks
Registered names, trademarks, etc. used in this document, even when not specifically marked as such,
are protected by law.
PerkinElmer is a registered trademark of PerkinElmer, Inc.
RamanMicro and Spectrum are trademarks of PerkinElmer, Inc.
Table of Contents
Introduction ............................................................................................... 5
About This Manual ............................................................................................ 6
Conventions Used in this Manual ........................................................................ 7
Notes, Cautions and Warnings ..................................................................... 7
Safety Practices ....................................................................................... 11
Overview .........................................................................................................12
General Operating Conditions............................................................................13
Fire Safety .......................................................................................................14
Electrical Safety ...............................................................................................15
Electrical Protection ...................................................................................15
Power Supplies ..........................................................................................15
Power Cords (Between PSU and Power Outlet Socket) .................................16
Fuses ........................................................................................................16
EMC Compliance ..............................................................................................17
Laser Safety .....................................................................................................18
Class 3B Safety Precautions ........................................................................19
General Laser Safety Issues .......................................................................20
Internal Interlock System ...........................................................................24
External Interlock System ..........................................................................24
Warning Labels ................................................................................................25
Labels on the Exterior of the Spectrometer .................................................25
Labels on the Exterior of the Microscope .....................................................26
Raman History, Theory and Instrumentation .......................................... 27
The History of Raman Spectroscopy ..................................................................28
Raman Theory .................................................................................................29
The RamanMicro 200........................................................................................31
The Czerny–Turner Spectrograph ...............................................................32
Preparing Your Facility ............................................................................ 33
Specifications ...................................................................................................34
Finding a Suitable Location for the Instrument ...................................................35
Instrument Dimensions .....................................................................................36
Installing Your RamanMicro 200 Series .................................................. 37
Unpacking the Instrument ................................................................................38
Lifting the Instrument .......................................................................................39
Attaching the External Interlock Override...........................................................40
Assembly of the Microscope ..............................................................................41
Fitting the Stage on the Microscope ............................................................41
Connecting the Motorized Stage .................................................................42
Attaching the Illumination Source ...............................................................44
Connecting the Ram an Module to the Microscope ........................................45
Connecting the Alignment Camera to the Raman Module .............................45
Connecting the Micr oscope to the Spectrometer Module ..............................46
Switching on the Instrument .............................................................................48
Installing the Instrument in Spectrum Software ........................................................49
Alignment of the Microscope .............................................................................50
Aligning the Microscope Objectives .............................................................50
Aligning the Raman Module ........................................................................51
Moving the Instr ument .....................................................................................56
Using the RamanMicro 200 Series ........................................................... 57
Spectrometer Front Panel .................................................................................58
Fiber Optic Probe .............................................................................................59
General Overview of the Fiber Optic Probe ..................................................59
Attaching and Removing the Fiber Optic Probe ............................................59
Acquiring Spectra with the Fiber Optic Probe ...............................................61
General SOPs .................................................................................................. 63
SOP-1: Turning the Instrument On ............................................................. 63
SOP-2: Turning the Instrument Off ............................................................ 65
SOP-3: Setting the Limits of the Motorized Stage ........................................ 66
SOP-4: Sample Positioning using a Manual Stage ........................................ 68
SOP-5: Sample Positioning using a Motorized Sample Stage ........................ 70
SOP-6: Setting-Up and Saving Experimental Parameters .............................. 73
Fiber Optic Probe SOPs .................................................................................... 76
SOP-7: Selecting the Fiber Optic Probe ....................................................... 76
SOP-8: Sample Positioning when using a Fiber Optic Probe ......................... 77
Calibration and Performance Verification SOPs .................................................. 78
SOP-9: Calibration/Performance Verificatio n ............................................... 78
SOP-10: Wavelength Calibration Verification ............................................... 79
SOP-11: Wavelength Calibration ................................................................ 81
SOP-12: Laser Frequency Calibration .......................................................... 82
SOP-13: Intensity Calibration ..................................................................... 83
Summary .................................................................................................. 85
Additional Parts and Upgrades .......................................................................... 86
Microscope Objective Lenses ..................................................................... 86
Sampling Accessories ................................................................................ 86
SERS Consumables ................................................................................... 87
Safety and Calibration ............................................................................... 87
Fiber Optic Probes ..................................................................................... 88
Maintenance and Troubleshooting .......................................................... 89
Maintenance .................................................................................................... 90
Cleaning the Spectrometer and Power Supply Units .................................... 90
Cleaning the Air Filters .............................................................................. 91
Servicing the Spectrometer ........................................................................ 92
Maintenance and Service of the Microscope ................................................ 92
Maintenance and Se rvice of the Fiber Optic Probe ....................................... 92
Replacing the Stage Controller Fuse .................................................................. 93
Troubleshooting .............................................................................................. 94
Hardware ................................................................................................. 94
Software................................................................................................... 94
Decontamination ............................................................................................. 96
WEEE Instructions for PerkinElmer Products ...................................................... 97
Introduction
6 . RamanMicro 200 Series Getting Started Guide
About This Manual
This Getting Started Guide contains all of the information that you will need to setup your
RamanMicro 200 Series Raman Microscope and start collecting spectra.
The Getting Started Guide is divided into the following chapters:
Introduction
This chapter conta ins an overview of the manual, and the conventions and warnings use d.
Safety Practices
This chapter outlines the electrical and laser safety issues. We recommend that all users rea d
this chapter.
Raman History, Theory and Instrumentation
This chapter conta ins a brief overview of the history and theory of Raman spectroscopy.
Preparing your Facility
This chapter shows you how to position your RamanMicro 200 Series instrument, and what
utilities are required, for example electricity and temperature control.
Installing your RamanMicro 200 Series
This chapter contains instructions about how to assemble your RamanMicro 200 system and
information on installing your instrument in the software.
Using the RamanMicro 200 Series
This chapter gives an overview of the instrument and contains an introduction to the Fiber
Optic Probe. It provides a number of Standard Operating Procedures (SOPs) that describe
how to perform certain tasks, for example how to collect a spectrum, or how to calibrate the
spectrometer.
Maintenance and Troubleshooting
This chapter conta ins informati on on how to maintain the system, and solutions to problems
you may encounter.
Introduction . 7
Conventions Used i n this M anual
Normal text is used to provide information and instructions.
Bold text refers to text that is displayed on the screen.
UPPERCASE text, for example ENTER or ALT, refers to keys on the PC keyboard. '+' is
used to show that you have to press two keys at the same time, for example, ALT+F.
All eight-digit numbers are PerkinElmer part numbers unless stated otherwise.
The term RamanMicro 200 Series means the RamanMicro 200 and the RamanMicro 200F
Raman Microscopes.
Notes, Cautions and Warnings
Three terms, in the following standard formats, are also used to highlight special
circumstances and warnings.
NOTE: A note indicates additional, significant information that is provided with some
procedures.
8 . RamanMicro 200 Series Getting Started Guide
einen Geräteschaden zu vermeiden.
undgå en beskadigelse af apparatet.
este tipo de circunstancias.
figurent dans un encadré semblable à celui-ci.
We use the term CAUTION to inform you about situations that could
CAUTION
result in serious damage to the instrument or other equipment. Details
about these circumstances are in a box like this one.
Caution (Achtung)
Bedeutet, daß die genannte Anleitung genau befolgt werden muß, um
Caution (Bemærk)
Dette betyder, at den nævnte vejledning skal overholdes nøje for at
Caution (Advertencia)
Utilizamos el término CAUTION (ADVERTENCIA) para advertir sobre
situaciones que pueden provocar averías graves en este equipo o en
otros. En los recuadros como éste se proporciona información sobre
Caution (Attention)
Nous utilisons le terme CAUTION (ATTENTION) pour signaler les
situations susceptibles de provoquer de graves détériorations de
l'instrument ou d'autre matériel. Les détails sur ces circonstances
Caution (Attenzione)
Con il termine CAUTION (ATTENZIONE) vengono segnalate situazioni
che potrebbero arrecare gravi danni allo strumento o ad altra
apparecchiatura. Troverete informazioni su tali circostanze in un
riquadro come questo.
Caution (Opgelet)
Betekent dat de genoemde handleiding nauwkeurig moet worden
opgevolgd, om beschadiging van het instrument te voorkomen.
Caution (Atenção)
Significa que a instrução referida tem de ser respeitada para evitar a
danificação do aparelho.
WARNING
Verletzung des Benutzers kommen kann.
overholdes.
dans un encadré semblable à celui-ci.
informazioni su tali circostanze in un riquadro come questo.
genomen, dit kan leiden tot verwondingen van de gebruiker.
We use the term WARNING to inform you about situations that could
result in personal injury to yourself or other persons. Details about
these circumstances are in a box like this one.
Introduction . 9
Warning (Warnung)
Bedeutet, daß es bei Nichtbeachten der genannten Anweisung zu einer
Warning (Advarsel)
Betyder, at brugeren kan blive kvæstet, hvis anvisningen ikke
Warning (Peligro)
Utilizamos el término WARNING (PELIGRO) para informarle sobre
situaciones que pueden provocar daños personales a usted o a otras
personas. En los recuadros como éste se proporciona información sobre
este tipo de circunstancias.
Warning (Danger)
Nous utilisons la formule WARNING (DANGER) pour avertir des
situations pouvant occasionner des dommages corporels à l'utilisate u r
ou à d'autres personnes. Les détails sur ces circonstances sont données
Warning (Pericolo)
Con il termine WARNING (PERICOLO) vengono segnalate situazioni
che potrebbero provocare incidenti alle persone. Troverete
Warning (Waarschuwing)
Betekent dat, wanneer de genoemde aanwijzing niet in acht wordt
Warning (Aviso)
Significa que a não observância da instrução referida poderá causar um
ferimento ao usuário.
10 . RamanMicro 200 Series Getting Started Guide
Safety Practices
12 . RamanMicro 200 Series Getting Started Guide
Overview
This chapter contains details of the safety precautions. Everyone who is going to install
and/or use the system must read and follow the precautions before unpacking, installing or
using the instrument.
Before performing any of the operations outlined in this manual, read all of the safety
practices and inst ructions, and make sure that you understand the procedures explai ned.
If you do not understand any of the details regarding general safety precautions, please
contact PerkinElmer before proceeding.
All information provided herein is for reference only. All information is believed to be correct
and complete. PerkinElmer shall not be liable for errors contained herein nor damages
concerned with the use of this document. PerkinElmer does not accept liability for the
interpretation of information resulting, either directly or indirectly, from our spectromete r s or
associated software interfaces. All results supplied by PerkinElmer’s spectrometers or
software are intended to provide initial determinations and results should be verified or
validated by using other appropriate techniques. All specifications and material contained in
this document are liable to change without notice. All safety information contained in this
document is superseded by governmental safety documents for your district or country; if in
doubt please consult American National Standards Ins titute’s Z136 series of laser safety
standards.
Safety Practices . 13
General Operating Conditions
If the equipment is used in a manner not specified herein, the protection
provided by the equipment may be impaired.
WARNING
The RamanMicro 200 Series spectrometers have been designed and tested in accordance
with PerkinElmer specifications and in accordance with the safety requirements of the
International Electrotechnical Commission (IEC). The RamanMicro 200 Series conforms to
IEC 61010-1 (Safety Requirements for electrical equipment for measurement, control and
laboratory use) as it applies to IEC Class 1 (earthed) appliances and therefore meets the
requirements of EC directive 2006/95/EC.
Only use the RamanMicro 200 Series spectrometers indoors and under the fol lowing
conditions:
Temperature 15 °C to 30 °C
Relative Humidity 80% maximum (non-condensing).
Avoid any adjustment, maintenance or repair of the opened, operating instrument. If any
adjustment, maintenance or repair of the opened instrument is necessary, this must be done
by a skilled person who is aware of the hazards involved.
The instrument has been designed to be safe under the following conditions:
• Indoor use
• Altitude up to 2000 m
• Ambient temperatures of 5 °C to 40 °C
• A maximum ambient relative humidity of 80% for temperatures up to 31 °C, decreasing
linearly to 50% relative humidity at 40 °C
• Mains fluctuations not exceeding ±10% of the nominal voltage.
Whenever it is likely that the RamanMicro 200 Series instrument is unsafe, make it
inoperative. The RamanMicro 200 Series may be unsafe if it:
• Shows visible damage
• Fails to perform the intended measurement
• Has been subjected to prolonged storage in unfavorable conditions
• Has been subjected to severe transport stresses.
14 . RamanMicro 200 Series Getting Started Guide
Fire Safety
• Ensure that there is at least 15 cm (6”) between any cooling vents and adjacent
equipment or walls.
• Ensure that the power supply unit (PSU) vents or fan do not become obstructed.
Safety Practices . 15
If the protective covers or service panels on the instrument appear to be
Electrical Safety
The spectrometer module comes assembled within a protective housing to prevent exposure
to electrical hazards. There is also a number of exterior service panels that you should
remove only if you are cleaning the air filters. For more information see
on page 91.
Filters
Removal of the protective housing or external panels, other than for
maintenance procedures specifically described in this manual, is
dangerous and is not recommended under any circumst ance s.
WARNING
damaged, turn the instrument off, secure the instr ument against a ny
unauthorized operation and conta ct PerkinElmer for advice before using
it again.
Electrical Protection
Insulation: Class I rating for external circuits. Only connect equipment that meets the
requirements of IEC 61010-1, IEC 60950 or equivalent standards.
Cleaning the Air
Power Supplies
The RamanMicro 200 is powered by one external PSU. The PSU used with the spectrometer
module can adjust automatically to a variety of AC power sources as outlined in Table 1.
Table 1 PSU input voltages and mains fluctuations information
Input Voltage Mains Fluctuations
100–230 V AC ±10% of the nominal voltage
Under no circumstances should you attempt to disassemble the PSU and
carry out repairs. If the PSU has a fault, please co ntact your PerkinElmer
Service Representative. The entire unit must be replaced.
WARNING
Connect the PSU to a three-wire (live, neutral and ground/earth) socket. The ground must be
a non-current carrying wire that is conne cted to the earth ground at the main distribution
box. The power point (socket) for the instrument PSU should be easily accessible for when
the instrument is serviced.
Check the line voltage and frequency of the power point (socket) before
the instrument is connected or the power is switched on. This should be
carried out prior to the installation of the instrument and at any time the
WARNING
instrument is plugged into a different power point (socket).
16 . RamanMicro 200 Series Getting Started Guide
Power Cords (Between PSU and Power Outlet Socket)
The power cord supplied to connect the PSU to the power point (socket) should be
appropriate for the country listed as the shipping destination.
If the power cord is not appropriate for your country it must b e removed
from use immediately and replaced.
WARNING
If the power cord has any physical damage it must be removed from use
immediately and replaced.
To avoid electrical hazards, the ground pin on the power cord should not
be interfered with in any way.
Fuses
The only fuses that you may have to replace are those in the mains power cords (UK-type
plug only) and the stage controller bo x fuse.
Safety Practices . 17
EMC Compliance
EC directive
The RamanMicro 200 and RamanMicro 200F have been designed and tested to meet the
requirements of the EC directive 2004/108/EC. The RamanMicro 200 Series complies with the
EMC standard EN61326 (EMC standard for electrical equipment for measurement, control
and laboratory use), and EN55011 (ISM) Class A (RF emissions).
FCC rules and regulations
This product is classified as a digital device used ex clusively as industrial, commercial, or
medical test equipment. It is exempt from the technical standards specified in Part 15 of the
FCC Rules and Regulations based on Section 15.103(c).
18 . RamanMicro 200 Series Getting Started Guide
Laser Safety
Details of the RamanMicro 200 Series laser source ar e summarized in Table 2.
Table 2 Laser Specifications
Class 3B
Wavelength 785 nm
Power output 250 mW (max)
The RamanMicro 200 Series complies with the requirements of IEC 60825-1:2007 and
21CFR 1040.10 FDA performance standards for laser products, except for deviations
pursuant to Laser Notice No. 50, dated 26 July 2001.
Use of controls or adjustments or the performance of any procedures
other than those specified in this document may result in exposure to
hazardous laser radiation.
WARNING
Removal of the protective housing or external panels is dangerous.
If the protective covers or service panels on the instrument appear to be
WARNING
damaged, turn the instrument off, secure it against any unauthorized
operation and contact PerkinEl mer for advice before using the instrument
again.
The RamanMicro 200 emits a focused beam of laser radiation at 785 nm (250 mW max.),
and is classified as a Class 3B laser system.
Avoid direct exposure to th e laser beam. Do not look at the laser light or
scattered laser light.
WARNING
During normal operation laser radiation is accessible from the microscope
objective lens. The output energy should be enclosed whenever possible
(using an enclosure or a beam attenuator) to avoid unnecessary exposure
WARNING
to laser radiation.
If you do not have an enclosure, you should use protective laser safety
goggles, with an optical density (OD) of at least 3 at 785 nm, as a
precaution against accidental exposure to the direct or reflected laser
light. Suitable goggles are available from PerkinElmer (part number
L1323518).
Safety Practices . 19
Class 3B Safet y Pr ecau ti ons
You should follow Class 3B laser safety preca utions such as the use of door interlocks and
the wearing of laser safety goggles. For information on these precautions refer to an
appropriately trained laser safety officer within your own facility. Laser safety is very
important. Inappropriate use of laser emitting devices may result in permanent injury. In the
United States the precautions are described in the AN SI Z136.1-20 00 Sta nd ar d. For
information on the ANSI Z136 standard please consult your laser safety officer and/or consult
http://www.z136.org. Outside the United States the guidelines are described in PD IEC TR
60825-14 (Safety of laser products – Part 14: A user’s guide). You should also be familiar
with the guidelines publi shed by the national authorities i n your country.
NOTE: In certain circumstances when the probe is permanently fixed in a closed
environment, for example inside a sealed reaction vessel, the device can be defined
as Class 1. Please refer to an appropriately trained laser safety officer within
your own facility.
NOTE: The RamanMicro 200 Raman Microscope can be fitted w ith an optional light-proof
enclosure, which is fitted with interlock sensors to ensure that the microscope cannot
be used with the enclosure doors open. The Ram an system is NOT reclassified to a
Class 1 instrument.
The foll o win g information is provided as a general overview only. This information is rudimentary
and is supplied to assist in the interpretation of this manual. Under no circumstances use this
informatio n as th e basi s of a laser safety code of behavior:
1. Class 3B lasers may emit visible and/or invisible radiation up to a maximum of 0.5 W
continuous wave (CW is longer than 0.25 s). They present a hazard to the eye if the direct
beam or spectral reflections are viewed without appropriate protection. You must only
operate Class 3B lasers in a designated area.
2. You must fit each Class 3B device wit h a captive key control switch, such that the key
cannot be removed from the lock except in the OFF position. The key should be
removed from the laser when it is not in use and kept in a safe place in the custod y of a
nominated person. Keys must never be issued to unauthor ized persons.
3. Each Class 3B device should be assessed by a suitably trained laser safety officer to
determine the accessible radiation emitted by the device. The laser safety officer should
consider laser power, wavelength and beam divergence when determining if the
Class 3B device’s remote interlock connector should be con nected to an interlock switch
at the entrance. For certain laser systems the laser safety officer may require the
shutter to be interlocked with the laboratory entrance in order to prevent inadvertent
exposure of persons entering to very hazardous laser radiation.
4. Rooms in which Class 3B devices are used should have notices affixed to the doors so
that they conform to the requirements of appropriate Laser Safety Regulations.
20 . RamanMicro 200 Series Getting Started Guide
General Laser Safety Issues
Laser Keyswitch
There is a captive laser key control switch on the front panel of the spectrometer. This key is
required to operate the laser. A laser safety officer or laboratory manager should co ntrol the
use of this key.
Laser Warning Labels
Laser warning labels are attached to the instrument for your safety. Please observe the
necessary precautions. For more details see
Warning Labels
Microscope Safety
The laser should also be turned off when the system is not in use. AVOID DIRECT
EXPOSURE TO THE BEAM.
on page 25.
Class 3B laser radiation
is emitted through the
objective lens
Figure 1 Schematic of the microscope
When using the microscope, the following precautions must always be observed:
• Never look directly into the laser beam or at the laser light scattered from a reflective
surface or sample.
• Avoid direct exposure to the laser light.
• Place warning signs near the laser operating areas.
• Control laser areas and limit access to individuals trained in the safe operation of lasers.
• Use protective laser safety goggles as a precaution to accidental exposure to the direct,
or reflected, laser light.
• Use an interlock switch on the door to the laboratory, if required.
WARNING
WARNING
Safety Practices . 21
Do not make any adjustments to the microscope or any part of the
system when experiments are in progress.
In particular:
•
Do not remove the objective lens from the microscope
•
Do not insert or remove samples
•
Do not disconnect the Raman module
•
Do not fit binocular eyepieces to the microscope module.
These adjustments may deflect laser light from t he safe light path
through the system.
Do not leave blank positions in the microscope turret when performing
your experiments. Ensure that all blank positions have protective covers in
position at all times.
If your system includes any Fiber Optic Probes , e nsure that protective
covers are in place when FOPs are not in use.
Fiber Optic Probe Safety
NOTE: This section is only applicable to RamanMicro 200F systems equipped with a fiber
optic probe.
The fiber optic probe body incorporates a number of important safety features, including:
• Beam attenuator/cover
• Emission indicator/LED.
When attached to t he spectrometer, the fiber optic probe provides a focused beam of
Class 3B laser radiation. During normal opera tion, laser radiation is accessible from the end
of the fiber optic probe. The LED emission indicator (Figure 2) at the probe head is
configured so that the LED lights up when the laser is turned on, indicat ing that laser
radiation is emitting from the probe aperture. The LED emission indicator is visible t hrough
laser safety goggles. The output energy should be enclosed whenever possible (using a
beam attenuator or cover) to avoid unnecessary exposure. The laser should be turned off
and the fiber optic probe attenuator cap replaced when the system is not in use. AVOID
DIRECT EXPOSURE TO THE BEAM.
22 . RamanMicro 200 Series Getting Started Guide
Probe body
Aperture label
Emission indicator
(Green LED)
Fiber optic cable
Beam attenuator / cover
Figure 2 Schematic of the fiber optic probe (not to scale)
When using the fiber optic probe, the following pre cautions should be observed:
• Never look directly into the laser beam or at the laser light scattered from a reflective
surface.
• Avoid direct exposure to the laser light.
• Use an interlock switch on the door to the laboratory and/or the reaction vessel.
• Place warning signs near the laser operating areas.
• Controlled access areas, limited to individuals tra ined in the safe operation of lasers, are
suggested for laser operations.
• Use protective laser safety goggles, with an OD (optical density) of at least 3 at 785 nm,
as a precaution against accidental exposure to the direct or reflected laser light. Suitable
goggles are available from PerkinElmer (part number L1323518).
• Ensure that the attenuator cap is attached.
Triggered Fiber Optic Probe Safety
When attached to the RamanMicro 200, a triggered fiber optic probe (TFOP) provides a
focused beam of Class 3B laser radiation. During normal operation, laser radiation is
accessible from the end of the probe.
The following precautions must be observed at all times:
• Keep the protective cover in place on the probe tip when the probe is not in use.
If you are using the TFOP with a plastic spacer, always rotate the spacer so that the
screw is beneath the probe barrel when not in use; when in this position the spacer acts
as a lens cap.
• Never look directly into the laser beam or at the laser light scattered from a reflective
surface or sample. Permanent eye-injury may result. Do NOT attempt to examine the
probe tip when the Path Active light on the probe is lit.
• Post warning signs near the laser operating areas.
Safety Practices . 23
2
1
.Π
Φ4
=
MPEb
f
NOHD
o
o
Φ
2
1
Φ
.442.
4
=
MPE
fNOHD
o
( )
( )
( )
[ ]
47.02
10101.10
−−
=
λ
MPE
bo
fo
• Controlled access areas, limited to individuals trained in the safe operation of lasers, are
suggested for laser operations.
• When the probe is in us e, always ensure that no stray laser radiation is directed at
people in the vicinity.
• Use protective laser safety goggles, with an optical density (OD) of at least 3 at 785 nm,
as a precaution against accidental exposure to the direct or reflected laser light. Suitable
goggles are available from PerkinElmer (part number L1323518).
• Do not use the TFOP if it shows any sign of damage.
• If repair or servicing is needed, call your PerkinElmer Service Representative for advice.
For further safety details, see the
Raman Triggered Fiber Optic Probe
leaflet (L1321940)
Calculation of Nominal Hazard Zones for use of Fiber Optic Probe
The following calculations show how to calcula te a nominal hazard zone. These calculations
are taken from ANSI Z136.1-1933. A laser safety officer uses these calculations to help
define how to use your Raman probe safely in your environment.
NOHD
Figure 3 Probe parameters used to calculate the nominal ocular hazard distance
The equation for a focused beam as given here (Figure B5 in ANSI Z136.1-1933) is used to
calculate the nominal ocular ha zard distance (NOHD) for the fiber optic probe. The equation
is:
Where:
fo = Probe focal length (cm)
b
= Diameter of laser beam incident on probe focusing lens (mm)
o
= Total radiant output power of probe (watts)
MPE =
Maximum permissible exposure (J cm−2)
For the FOP bo is 0.254 cm, therefore the above equation can be simplified to give:
For the wavelength range 700–1050 nm the MPE for a 10 second exposure can be calculated
from:
−2
J cm
24 . RamanMicro 200 Series Getting Started Guide
Where λ is the laser wavelength in micrometers (µm). For example, a laser wavelength of
785 nm (0.785 µm) will have an MPE value of 1.49 x 10
a 10 second exposure or longer . Table 3 shows the calculated nominal ocular hazard distance
for 785 nm Raman systems.
Table 3 Nominal Ocular Hazard Distance (NOHD) for 785 nm
Probe focal length / mm
7.5 (± 10%)
NOTE: Other focal lengths are available; contact PerkinElmer for details. In particular, the
use of an ultra-long working distance lens adaptor (part number L1320071) provides
a working distance of 25 mm. Where different focal lengths are used, the NOHD
should be calculated accordingly.
NOHD for 100 mW exiting fiber optic probe / cm
−2
J cm−2 or 1.49 x 10−3 W cm−2 for
27.3
Internal Interlock System
There is no internal interlock system fitted to the RamanMicro 200. If instrument panels are
removed collimated laser light may be visible. Under no circumstances remove any
instrument panels .
External Interlock System
Class 3B laser systems have the potential to cause harm or injury to those using them or
others in the vicinity of the instrument. For more infor mation refer to
Interlock
on page 40.
Attaching the External
Warning Labels
MAXIMUM OUTPUT: 250mW
PULSE DURATION: CONTINUOUS WAVE
EMITTED WAVELENGHS: 785nm
THIS LASER PRODUCT CONFORMS TO
IEC 60825-1: 1993, 21 CFR 1040.10 & 1040.11
LASER RADIATION
AVOID EXPOSURE TO BEAM
CLASS 3B LASER PRODUCT
MAXIMUM OUTPUT: 250mW
PULSE DURATION: CONTINUOUS WAVE
EMITTED WAVELENGHS: 785nm
THIS LASER PRODUCT CONFORMS TO
IEC 60825-1: 1993, 21 CFR 1040.10 & 1040.11
LASER RADIATION
AVOID EXPOSURE TO BEAM
CLASS 3B LASER PRODUCT
CAUTION – CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM
CAUTION – CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM
When this label is attached to an instrument it means refer to the
manual to find out the nature of the pot ential hazard and any actions
which have to be taken.
Labels on the Exterior of the Spectrometer
Power Max. 100W
DC Voltage 5V 12V
Current Max. 4A 6.5A
Safety Practices . 25
Figure 4 Labels on the spectrometer module
26 . RamanMicro 200 Series Getting Started Guide
LASER APERTURE
CAUTION – CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM
CAUTION – CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM
Labels on the Exterior of the Microscope
Figure 5 Labels on th e mic ros c o pe
Raman History, Theory
and Instrumentation
28 . RamanMicro 200 Series Getting Started Guide
The History of Raman Spectroscopy
Inelastic light scattering was first predicted in 1923 by A. G. Smekal in Germany. Research
carried out by A. H. Compton discovered inelastic scattering of X-rays, which led the Indian
scientist C. V. Raman to investigate this phenomenon. Raman and his assistant Krishnan
observed what came to be known as the Raman effect (Figure 6) for the first time in 1928.
They separated the blue light from sunlight and observed the scatteri ng from water and
alcohol visually. In addition to the blue incident light, a faint green light also scattered, later
named Stokes scattering. By the 1930s much o f the theory unde rlying Raman scattering was
understood, but due to the complexity of the instrumentation the initial interest in Raman
spectroscopy was not maint ained. Improvements in charge-coupled device (CCD) detectors,
filters and lasers over the past 25 years have radically simplified Raman instrumentation, and
interest in Raman spectroscopy has been renewed. Lasers are ideal excitation sources for
Raman because they give highly monochromatic radiation and provide high irradiances at the
sample.
Raman History, Theory and Instrumentation . 29
Laser Photons in
Lens
Raman Theory
When photons from a monochromatic light sour ce impinge on a sample, a small fraction are
scattered in all directions. Some of the scattered photons are of the same energy as those of
the incident beam, and hence the sa me frequency, giving rise to Rayleigh scattering
(Figure 6).
Sample
No change in energy,
no useful information
Loss of energy spectra
Photos gain energy,
but signal is very
small (anti-Stokes)
Intensity
Figure 6 The Raman effect: irradiation of sample and subsequent scatter
Some photons will be inelastically scattered and w ill be detected at either longer wavelengths
(lower frequencies, ν’, Stokes scattering) or shorter wavelengths (higher frequencies, ν’’,
Anti-Stokes scattering). With Stokes scattering, some of the incident photons give up a
portion of energy to the molecules and are detected at a lower frequency (longer
wavelength) than the Rayleigh radiation (Figure 7).
(Stokes)
Excited Energy
Rayleigh
Stokes
Figure 7 Rayleigh, Stokes and Anti-Stokes scattering
Virtual Energy State
Anti-Stokes
30 . RamanMicro 200 Series Getting Started Guide
Anti-Stokes x12
Rayleigh
Raman Shift (cm−1)
The change between the incident radiation and the Stokes/Anti-Stokes radiation represents a
change in the vibrational and/or rotational energy of the molecule. The frequency shifts from
the frequency of the incident beam are displayed on the x-axis of a Raman spectrum,
generally in relative wavenumber s (otherwise known as Raman Shift) and the intensity of the
bands are displayed in arbitrary units on the y-axis.
Stokes
Figure 8 Raman spectrum of cyclohexane, showing Stokes, anti-Stok es a nd
Rayleigh scattering
Raman History, Theory and Instrumentation . 31
Microscope frame
The RamanMicro 200
The RamanMicro 200 is a Raman microscope system with laser excitation wavelength at
785 nm.
Alignment camera
Raman module
Illuminator
module
Objectives
Stage
Figure 9 The RamanMicro 200 microscope
The microscope (Figure 9) is composed of a frame, illumination module, Raman module and
camera module. This is connected to the spectrometer module by two fiber optic cables. The
spectrometer module contains the spectrograph and CCD detector. The laser supplies light
through an objective lens onto the sample. The scattered light from the sample is collected
with the same objective lens and transmitted back to the spectrometer. A schematic of the
system configuration is provided in Figure 10.
Optical Head
Laser
Spectrograph
CCD
detector
Figure 10 Schematic of the RamanMicro 200 system
32 . RamanMicro 200 Series Getting Started Guide
The Czerny–Turner Spectrograph
The RamanMicro 200 is equipped with a Czerny–Turner spectrograph. The Czerny–Turner
spectrograph is the most commonly used spectrograph design in Raman spectroscopy. Here,
light is focused into the spectrograph, collimated and shone on a grating which disperses
that light in one dimension. The light is then focused onto a photo-sensitive CCD detector
(typically 1024 p ixels wide), which detects the Raman spectrum.
CCD
Detector
Lens #2
Entrance
Slit
Lens #1
Grating
Figure 11 Schematic of the Czerny-Turner spectrograph
The Czerny–Turner spectrograph is equipped with a fixed grating, and the range of light
falling on the CCD can be factory configured to be approximately 95–3200 cm
resolution of about 8 cm
−1
.
−1
at a
Preparing Your Facility
34 . RamanMicro 200 Series Getting Started Guide
Specifications
Weight 20 kg 20 kg 2.8 kg
Height 161 mm 520 mm 90 mm
Width 530 mm 370 mm 220 mm
Depth 600 mm 430 mm 184 mm
Power Supply 100–230 V ±10%,
Primary Fuse – See Olympus manual 1.6 A T (time-lag)
Laser Class Class 3B –
Operational
Temperature
Recommended
Temperature
Spectrometer Microscope Stage Controller
(for motorized XYZ
stage only)
50–60 Hz
100–120 V/220–240 V
50–60 Hz
15 °C – 30 °C
20 °C – 25 °C
100–240 V,
50–60 Hz
If the electricity supply does not conform to these specifications, please consult PerkinElmer
prior to installation. If the supply of power is erratic please use an uninterruptible power
supply (UPS). Incorrect shut down, power fluctuations or brown-outs may damage the
spectrometer.
Preparing Your Facility . 35
Finding a S uitabl e Loc atio n for the Instrument
The following conditions should be observed:
• The environment must be suitable for a Class 3B laser system.
• There must be a minimum of 4 power points (1 for the microscope, 1 for the
spectrometer and 2 for the PC). If your system has a motorized stage you will also
require 1 power point for the stage controller.
• The workbench must be flat and level.
• The workbench where the spectrometer and PC are to be located should be at a
suitable height to allow the user to work comfortably.
• Use conducting floor coverings to minimize static discharge.
• Floor vibrations or noise from heavy manufacturing equipment can affect the
performance of t he instrument.
• Intense magnetic fields should be at least 5.5 meters (18 feet) away from the
spectrometer.
• Never place the instrument near a window that gets direct sunlight.
Ensure all users understand laser safety issues before locating and using the instrument.
Consult your Laser Safety Officer and perform a risk assessment before installing the
equipment.
36 . RamanMicro 200 Series Getting Started Guide
600 mm
520 mm
Instrument Dimensions
The RamanMicro 200 spectrometer module weighs approximately 20 kg. The microscope
module weighs approximately 20 kg. Laboratory benching must be strong enough to support
the combined weight of the microscope, spectrometer module and any ancillary equipment
without warping or wobbling. The be nch must be at least 610 mm (26”) in depth.
161 mm
530 mm
370 mm
Figure 12 Dimensions of the RamanMicro 200 system
430 mm
Installing Your
RamanMicro 200 Series
38 . RamanMicro 200 Series Getting Started Guide
Unpacking the Instrument
When the instrument has been delivered, please move it with care and avoid shaking or
dropping the spectrometer. Allow the instrument to equilibrate to room temperature for at
least 12 hours before opening. The instrument may have been stored at low temperature
prior to delivery and you should allow it to equilibrate to stop moisture condensing on the
internal electronics.
When unpacked, check that you have the following parts and check for any physical damage.
If there is any damage, do not continue with the installation. Contact your PerkinElmer
Service Representative immediately.
Part Number Description
LX108870 Spectrum Std for Raman Software Kit
L1323503 Laser key (x2)
L1321802 Mains PSU
L1320821 USB Cable
L1320264 Ext ernal Interlock Override
L1323055
OR
L1323011
L1865749 RS232 Ca b le (systems with motorized stages only)
L1860289 Stage Controller (systems with motorized stages only)
RamanMicro 200 Series
Contains the microscope, the Raman module, the alignment camera
module, and the spectrometer module.
Contains the Spectrum Std Software CD (LX100877), the Raman
Instrument Configuration Disk (LX108869), the Raman Spectral Libraries
CD (L1321877) , and the Spectrum Insight So ftware CD (LX108872)
Manual Stage
Motorized Stage
L1865723 Joystick and cable assembly (systems with motorized stages only)
L1181228 Polystyrene Sample
LX108897 SpectrumIMAGE Software Kit (systems with motorized stages only)
N1870151 Microscope Tools Kit
NOTE: Your instrument is intended for installation by a PerkinElmer Service Representative.
There are no user-serviceable parts in the RamanMicro 200 Series. The following
sections are provided for your informa tion only.
Installing Your RamanMicro 200 Series . 39
Lifting the Instrument
The RamanMicro 200 spectrometer and microscope modules each weigh 20 kg. To avoid
injury or damage to t he instrument, please use proper lifting techniq ues.
To lift and/or move the instrument safely requires two people.
WARNING
WARNING
Always hold the instrument under the base when you lift the
spectrometer.
40 . RamanMicro 200 Series Getting Started Guide
Attaching the Exte rn al Inte rl ock Override
The RamanMicro 200 Series has an external int erlock connection located on the rear panel of
the spectrometer module. To operate the instrument, the external interlock must be
satisfied.
You must connect an external door inte rlock to the 7-way DIN connec tor labeled
INTERLOCK on the rear panel of the spectrometer, or attach the External Interlock Override
(L1320264) supplied with the instrument.
If your laser safety officer determines that an interlock is required on the door to the room,
you will need to purchase the Raman Laser Safety Kit (L1320262). For more information
contact your PerkinElmer representative.
Installing Your RamanMicro 200 Series . 41
Assembly of the Microscope
The Olympus BX51 microscope is designed to enable you to analyze microscopic samples.
You can purchase a manual stage or a motorized stage.
To prevent potential injury to yourself and damage to the instrument,
switch OFF all components in the system and disconnect them from the
line power supply before you perform any of the procedures described
WARNING
Fitting the Stage on the Microscope
To fit the stage:
1. Unpack the stage from the shipping material.
2. Remove the objective nosepiece from the microscope by unfastening the grub screw on
the right-hand side of the nosepiece.
in this section.
3. Carefully locate the stage above the stage support ring and then lower the stage into
position.
4. Ensure that the stage is level, and then fasten the stage in position by tightening the
screw at the front o f the stage (Figure 13).
Fixing screw
Figure 13 Fitting the stage (manual stage shown)
5. Replace the objectives and tighten the securing grub screw.
If you have a manual stage, you can upgrade to a motorized stage at any time. For more
information contact your PerkinElm er representative.
42 . RamanMicro 200 Series Getting Started Guide
Connecting the Motorized Stage
The motorized stage and joystick (L1865723) are powered by the stage controller box
(L1860289).
Figure 14 Stage controller and joystick
To connect the motorized stage:
1. Connect the cable from the joystick to the socket labeled JOYSTICK on the rear panel
of the stage controller box (Figure 15).
Figure 15 Rear panel of the Stage Controller
2. Connect the cable from the socket on the right side of the stage to the MOTOR X and
MOTOR Y sockets on the rear panel of the stage controller box.
3. Connect the RS232 cable (L1865749) from the PC to the socket labeled RS232 on the
rear panel of the stage controller.
4. Connect the power cable from the stage controller box to the mains supply.
Installing Your RamanMicro 200 Series . 43
Attaching the Z-axis Focus Motor and Sleeve
NOTE: If you have purchased the optional microscope light-proof enclosure, attach the z-
axis focus motor and sleeve before fitting the enclosure. See the
Light-Proof Enclosure
1. Slide the focus motor sleeve over the z-axis drive control knob on the left side of the
microscope, and then tighten the co nnections using a hexagonal key.
2. Push the focus motor into the sleeve until it touches the drive control , and then tighten
the screw to finger tight.
3. Connect the serial connector from the focus motor int o the connection at the rear o f the
stage control box labeled MOTOR Z (Figure 15).
leaflet (L1321873) for more information.
Raman Microscope
Figure 16 Z-axis focus motor and sleeve attached to the microscope inside the
optional light-proof enclosure
NOTE: Ensure that the pre-focusing lever on the coarse adjustment control is on the off
position.
44 . RamanMicro 200 Series Getting Started Guide
Screws
Power cable
Allen
Attaching the Illumination Source
To attach the illumination source to the illuminator module:
1. Hold the illumination source in position against the rear of the illuminator module.
2. Using the allen key supplied (Figure 17), fasten the two screws so that the source is
supported.
3. Connect the black cable from the illumination source to the microscope frame.
Top View
key
Figure 17 Attaching the illumination source to the microscope
Installing Your RamanMicro 200 Series . 45
Alignment camera
Mounting bracket
Raman
Connecting the Raman Module to the Microscope
The Raman module is positioned on top o f the illuminator module so that the connecter
bracket is aligned above the mounting hole. Ensure that the side of the Raman module is
aligned with the illuminator module and then tighten the grub screw.
module
Illuminator
module
Figure 18 Connecting the Ra ma n module
Screw
Connecting the Alignment Camera to the Raman Module
The alignment camera module is then connected into the mounting bracket on top of the
Raman module, and the screws tightened.
NOTE: Do not attach more than two alignment cameras in the Raman system.
Screw
Figure 19 Connecting the alignment camera
NOTE: Do not connect the USB from the camera to the PC until after the software is
installed. See
Installing the Instrument in Spectrum Sof tw a re
on page 49.
46 . RamanMicro 200 Series Getting Started Guide
Connecting th e Microscope to the Spectrometer Module
The microscope is connected to the spectrometer module by two fiber optic cables. The
excitation fiber transmits the laser energy from the spectrometer to the microscope, and the
collection cable transmits the collected Raman data back to the detector.
1. Connect the excitation cable (L1323331) from the Raman module to the EXC coupler
under LED FIBER 1 on the rear panel of the spectrometer.
Ensure that the end of the fiber with the white cable marker is attached to the Raman
module.
Try to bend the fiber as little as possible. If you bend the fiber through a
CAUTION
tight angle it can be damaged.
Excitation fiber
Collection fiber
Figure 20 Fiber optic cables con necting the Raman module to the spectrometer
Installing Your RamanMicro 200 Series . 47
Coupler
Make sure that the polarizing ridge on the fiber optic cable is aligned with the slot in the
coupler casing before carefully pushing the cable into the fiber bracket.
Polarizing ridge
aligned with slot
Fiber optic cable
Figure 21 Connecting the fiber optic cable
2. Connect the collection cable (L1323332) from the Raman module to the COL coupler
under LED FIBER 1 on the rear panel of the spectrometer.
Ensure that the end of the fiber with the white cable marker is attached to the Raman
module.
Make sure that the polarizing ridge on the fiber optic cable is aligned with the slot in the
coupler casing before carefully pushing the cable into the coupler (Figure 21).
Try to bend the fiber as little as possible. If you bend the fiber through a
CAUTION
tight angle it can be damaged.
3. When you have connected both cable s, ensure that t hey do not touch the illumination
source.
You could use a cable tie to direct t he cables round the source.
48 . RamanMicro 200 Series Getting Started Guide
Switching on the Instrument
NOTE: Do not connect the USB from the alignment camera to the PC until after the software
is installed. See
1. Insert the key into the laser power keyswitch on the front panel of the spectrometer
module and ensure that it is turned to the ARM position.
The RamanMicro 200 is supplied with two keys. One should be placed in the instrument
and the other should be stored in a safe place. Replacement keys (L1323503) can be
obtained from PerkinElmer. A laser safety officer or lab manager can control use of the
instrument with th ese keys.
2. Connect the instrument power supply unit (L1321802) to the RamanMicro 200
spectrometer Main Power
Do not connect to the Aux Power connector.
3. Connect the power cable from the microscope to the mains supply (Figure 17), and then
switch on the microscope.
4. If you have a motorized stage, switch on the stage controller box (Figure 15).
Installing the Instrument in Spectr um Softw a re
socket and then switch on.
on page 49.
Installing Your RamanMicro 200 Series . 49
Installing the Instrument in Spectrum Software
You must not connect the instrument USB cable or the microscope alignment camera USB
cable to the PC before the Spectrum software has been installed.
NOTE: A PerkinElmer Service Representative will install the Spectrum software on the PC
and add set up your instrument in the software. However, details of the software
installation can be found in the
L1050095).
After the softwa re is installed, you should switch on your Raman instrument. Wait a few
moments for your instrument to initialize and then c onnect the microscope alignment camera
USB cable to the PC.
The instrument then needs to be added to the software. See “Installing a Raman
Instrument” in the
multiple FT-IR spectrometers to Spectrum on the same PC as a Raman instrument. However,
you can only add one Raman instrument to Spectrum.
You should now continue to connect the fiber optic cables for any fiber optic probe or
triggered fiber optic probe to the RamanMicro 200 Series instrument, as described in
Spectrum Administrator’s Guide
Attaching and Removing the Fiber Optic Probe
Spectrum Administrator’s Guide
(part number L1050095). You can add
on page 59.
(part number
NOTE: For details of how to use a triggered fiber optic probe, see
Optic Probe User’s Guide
(part number L1321940).
Raman Triggered Fiber
50 . RamanMicro 200 Series Getting Started Guide
Alignment of the Microscope
Do not disconnect the microscope from the spectrometer. Only trained
PerkinElmer Service Representatives should attach and remove the fiber
optic cables that connect the two modules.
WARNING
The RamanMicro 200 system consists of four primary components:
• The spectrometer module (containing the laser and the detector)
• The microscope
• The Raman module
• The alignment camera module.
Please read and understand the general safety precautions before proceeding. If you do not
understand any of the details regarding general safety precautions, please contact
PerkinElmer before performing any of the following procedures.
Aligning the Microscope Objectives
NOTE: Do not touch the glass of the objective lenses when you select the appropriate
magnification. Do not push on an objective to rotate the nosepiece, but use the
revolving plastic ring.
The microscope objective nosepiece can fit up to 4 objectives.
1. Select the highest magnification objective lens on the microscope (either the 100x or
50x) by rotating the plastic ring on the objec tive nosepiece.
2. Select the Bright Fi eld (BF) switch position on the microscope.
3. Place a slide with an easily recognizable feature on it (for example, a graticule slide).
4. In the Spectrum software, select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
5. On the Setup Instrument Basic tab, ensure that Microscope or Microscope XYZ
Stage, as applicable, is selected as the Accessory.
6. Select the Setup Microscope or Setup Microscope XYZ Stage tab, as applicable, and then
select Live Video.
7. Move the stage unti l the feature is in focus and in the center of the crosshairs in the
Live Video.
Ensure that the image remains sharp and focussed.
8. Select the next highest magnification objective.
Installing Your RamanMicro 200 Series . 51
9. Insert the adjuster keys supplied with the microscope into the nosepiece and move the
adjusters until the crosshairs are aligned with the same feature as for the previous
objective lens.
Figure 22 Microscope objective adjusters
10. Repeat Steps 8–9 until all the objectives are aligned.
Figure 23 Feature in center of cross hairs at three objective magnifications
Aligning the Raman Module
There are three levels of optical alignment in the Raman module:
(i) Alignment of the laser path
(ii) Alignment of the video camera
(iii) Alignment of the Raman coupler
Level (ii) and level (iii) may be carried out without removing any instrument paneling. Level
(i) requires removal of instrument paneling and must only be carried out by a PerkinElmer
Service Representative, and in a Class 3B environment.
Aligning the Laser Path
The laser path only needs realignment if the beam is observed to move grossly in the
xy
-plane when moving into and out of f ocus. If the laser path is aligned correctly, then the
laser spot should get larger in off-focus positions without appreciable lateral movement.
NOTE: The requirement for laser alignment is NOT indicated by a situation where the laser
spot and the center of the crosshairs are non-coincident (this scenario requires
realignment of the video camera with the laser spot).
52 . RamanMicro 200 Series Getting Started Guide
If the system is misaligned, we strongly recommended that you contact a PerkinElmer
Service Representative. Changes to the laser path alignment will have a dramatic impact on
the performance of your instrument.
NOTE: Movement of the laser spot in the xy-plane may be caused if the surface of the
sample is not flat or not mounted correctly. Ensure that the stage is level and that
the sample is flat. PerkinElmer recommends the use of a flat silicon surface.
Aligning the Video Camera with the Laser Spot
The crosshairs displayed on the Setup Microscope or Setup Microscope XYZ tab in the
Spectrum software represent the position of the laser. If the microscope is knocked, then this
crosshair may not correspond to the true position of the laser. Minor corrections of the
crosshair position can be made in the software using Align Video with Laser. If the laser
spot is very far from the crosshair, then a complete realignment of the video camera is
required:
1. Select the 20x objective lens on the microscope.
2. Place a flat sample on a microscope slide.
We recommend that you purchase the s CAL Raman Calib ration Standard (L 1321843) for
this alignment procedure.
We recommend that you do not use a colored polymer sample.
3. In the Spectrum software, select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
4. On the Setup Instrument Basic tab, ensure that Microscope or Microscope XYZ
Stage, as applicable, is selected as the Accessory.
5. Ensure that the correct objective magnification is selected in the Microscope Objective
drop-down list.
6. Select the appropriate Sample Holder in the drop-down list and load the sample holder
onto the sample stage, following the instructions on-screen if you have a motorized
stage.
7. Select the Setup Microscope or Setup Microscope XYZ Stage tab, as applicable, and then
select Live Video.
8. Focus on a flat, non-reflective area of sample, and then select the Dark Field (DF)
switch position on the microscope and switch off the illumination.
9. Select Toolbox and then select Align Video with Laser from the sub-menu.
The Align Video with Laser wizard starts.
Installing Your RamanMicro 200 Series . 53
10. Ensure that you have followed the instructions on the screen, and then click Next.
The following dialog is displayed.
11. Adjust the Laser Power until you can see a clear visible spot.
12. If the laser spot is visible and close to the cross ha ir s, you do not need to physically
adjust the camera module. Continue at step 16 to perform a fine adjustment to align
the cross hairs with the laser spot.
OR
If no spot is visible, a physical adjustment of the camera module is required. Continue
at step 13.
13. Loosen the 4 screws on t he top plate of the Raman module until the camera is just free
to move (Figure 24).
Figure 24 Top view of the microscope showing the screws used to adjust the
position of the camera module
14. Move the camera module until the crosshairs line up as closely as possible with the
center of the laser spot.
15. Re-tighten the 4 screws.
54 . RamanMicro 200 Series Getting Started Guide
16. Click Next.
The following dialog is displayed.
17. Use the arrows in the Align Video with Laser dialog to f ine -tune the position o f the
crosshairs to be in the center of the laser spot.
You will need to repeat the fine adjustment of the cross hairs using the Align Video with
Laser function for each mic roscope objective, but do not adjust the camera module.
Aligning the Raman Coupler
Adjustments to the Raman coupler can have a dramatic impact on the
CAUTION
performance of your Raman instrument. We recommend that you
contact your PerkinElmer Service Representative.
You should only adjust the Raman coupler if the signal throughput has decreased. This may
xy
occur if you have moved the unit. The only adjustment to the Raman is the
the rear of the Raman module (see Figure 25). The procedure for alignment is as follows:
1. Place a flat sample of silicon on the sample stage.
We recommend that you purchase the s CAL Raman Calib ration Standard (L 1321843) for
this alignment procedure.
2. Loosen the 4 retaining screws at the coupler so that abo ut 0.5 mm of thread is
showing.
3. Ensure that the Bright Field (BF) switch position is selected on the microscope, and that
the illumination is t urned on.
4. In the Spectrum software, select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
5. On the Setup Instrument Basic tab, ensure that Microscope or Microscope XYZ
Stage, as applicable, is selected as the Accessory.
-adjuster at
6. Select the appropriate Sample Holder in the drop-down list.
7. Select the lowest magnification objective on the microscope.
8. Ensure that the correct objective it is selected in the Microscope Objective drop-down
list on the Setup Instrument Basic tab.
9. Select the Setup Microscope or Setup Microscope XYZ Stage tab, as applicable, and then
select Live Video.
Installing Your RamanMicro 200 Series . 55
10. Ensure that the sample is in fo cus.
11. Repeat steps 7–10 for each objective until the highest magnification is selected.
12. Switch to the Dark Field (DF) switch position on the microscope, and turn off the
illumination.
13. Cover the microscope with a dark cloth or, if fitted, close the light-proof enclosure doors
and ensure that caps are placed on any external fiber optic probes or accessories, to
minimize stray light from interfering with the calibration.
14. Click .
The RamanMicro 200 will start collecting data in the Live display.
15. Observe the data displayed in the Live display.
16. Very slowly mo ve one of the coupler knobs clockwise and counter-clockwise until the
Raman signal is maximized.
Figure 25 Adjustment of the XY Raman coupler
17. Move the other knob in a similar manner until the Raman signal has maximized.
18. Return to the first knob and move again to maximize the Raman signal (the movement
should be much less than used the first time).
19. Return to second knob and do likewise.
xy
20. Tighten each retaining screw a little and move the
each time.
21. Tighten screws until the plate at the rear coupler is secure – do not over tighten.
-coupler knobs to compensate
56 . RamanMicro 200 Series Getting Started Guide
Moving the Instr ument
The RamanMicro 200 spectrometer weighs about 20 kg. The
RamanMicro 200 microscope module weighs about 20 kg. Two people
are needed to lift the instrument.
WARNING
Please contact PerkinElmer for advice about removing the fiber optic cables that connect the
spectrometer and microscope modules. If you need to ship your instrument you must use the
original supplied packing materials to prevent damage to the system.
When the instrument has reached its destination and has been reassem bled, you should
perform the appropriate calibration/verification procedures as described in the
and Performance Verification SOPs
on page 78.
Calibration
Using the
RamanMicro 200 Series
58 . RamanMicro 200 Series Getting Started Guide
Spectrometer Fro nt Pa nel
The front panel of the spectrometer module contains 3 LED indicators and 1 key switch
(Figure 26).
Figure 26 Spectrometer module front panel
LED Description
Power If the Power LED is illuminated, this indicates that the instrument is receiving
power.
Laser This LED indicates the power status of the laser . If the LED is illuminated then
the laser is receiving power. This does not mean that laser radiation is present
throughout the optical path of the instrument, only that the la ser power is on.
Active This LED indicates the status of the laser shutters. If the LED is illuminated then
the shutters may be activated (open) and laser radiation present throughout the
optical path of the instrument.
NOTE: If you open the microscope light-proof enclosure (if fitted) when the Active LED is
illuminated, the safety interlocks will close the shutt ers.
NOTE: The shutter on the RamanMicro 200 instrument will be open continuously during
acquisition. Care should be taken when scanning and monitoring not to damage your
sample.
Using the RamanMi cro 200 Series . 59
Fiber Optic Probe
NOTE: This section is only applicable if you have a RamanMicro 200F that is fitted with a
fiber optic probe.
NOTE: For details of how to use a triggered fiber optic probe, see the
Fiber Optic Probe
leaflet (L1321940).
Raman Triggered
General Overview of the Fiber Optic Probe
The fiber optic probe (FOP) is designed to enable the remote measurement of samples. It is
available in a range of different configurations, depending on the end-user’s application.
However, it should be noted that compatibility of the working environment with the probe
should be verified with PerkinElmer prior to use.
The probe body contains the filtering optics and is interfaced with two fiber optic cables (an
excitation fiber and collection fiber). Care should be taken when handling the probe body so
that the optical window is not soiled or damaged.
NOTE: If you use the probe body in any chemical or aggressive environment you must use
an immersion sleeve to protect the probe body. DO NOT ATTEMPT TO USE THE
PROBE BODY IN SUCH AN ENVIRONMENT WITHOUT THE USE OF AN
IMMERSION SLEEVE (L1320003). If required, contact PerkinElmer for further
details.
Attaching and Removing the Fiber Optic Probe
The standard fiber optic probe is constructed such that a single armored cable bifurcates into
an excitation cable and a collection cable. The excitation fiber transmits the laser energy,
whereas the collection cable is a conduit for the Raman energy. Both fibers terminate in FC
style connectors. If you hav e a standard probe , there is also a DIN connector attached to the
end of two wires w hich connects the fiber optic LED to the instrument. If you have a
triggered probe, there is an Ethernet cable that connects to a USB extender which connects
the probe to the PC.
Attaching the FOP
1. Ensure the attenuator cap is attached to the FOP.
2. Turn off the instrument and disconnect the main power cable and, if appropriate, the
stage controller power cable.
3. Carefully move the spectrometer module to permit easier access to the rear panel.
4. Remove the two retained screw-in caps from the two coupler ports marked EXC and
COL below the FIBER 2 connector on the rear panel (Figure 27).
60 . RamanMicro 200 Series Getting Started Guide
Figure 27 Attaching the FOP
5. Connect the excitation cable of the fiber optic probe to the coupler marked EXC.
Make sure that the polarizing ridge on the fiber optic probe cable is aligned with the slot
in the coupler casing before carefully pushing the cable into the coupler (Figure 28).
Coupler
Probe
Polarizing ridge
aligned with slot
Figure 28 Connecting the fiber optic probe
6. Connect the collection cable of the fiber optic probe to the coupler marked COL.
Make sure that the polarizing ridge on the fiber optic probe cable is aligned with the slot
in the coupler casing before carefully pushing the cable into the coupler.
7. Screw the screw caps from the fiber optic cables onto the coupler casings.
8. If you have a standard probe, connect the DIN connector to the nearby socket marked
LED FIBER 2.
OR
If you have a triggered probe, connect the USB extender cable to a USB port on your
PC.
9. Reconnect the p ower cables.
Removing the FOP
If it is necessary to remove the FOP, for example, for replacement or transportation, please
follow this procedure carefully:
1. Ensure the attenuator cap is attached to the FOP.
2. Switch off the instrument and remove the main power cable.
3. Carefully move the spectrometer module to permit easier access to the rear panel.
Using the RamanMi cro 200 Series . 61
4. If you have a standard probe, remove the DIN connector in the socket marked
LED FIBER 2.
OR
If you have a triggered probe, disconnect the USB extender cable from the USB port on
your PC.
5. Carefully remove the retained ends of the FOP by unscrewing and pulling the cables
away from the couplers perpendicularly.
Be extremely careful when removing the FOP as you can easily damage the instrument
and the fiber optic cables.
6. Replace the protective plastic caps on the fiber ends of the FOP that have been
removed from the rear panel.
7. Fasten the retained screw-in caps onto the couplers on the rear panel.
8. Reconnect the p ower cables.
Acquiring Spectra with the Fiber Optic Probe
The Raman fiber optic probe has a tight focus spot that is approximately 7.5 mm away from
the lens. The focused spot enables you to pinpoint the measurement area on the sample.
Sampling Solids
Solids can be analyzed by mounting the probe on a stand and adjusting the focal spot on the
surface of the sample. Whenever possible, powders should be compacted down and thin
films of materials should be folded so that the samp le covers the entire depth of field. Solids
can be analyzed in glass and plastic containers, similar to liquid samp les.
NOTE: Raman spectra are measured in the visible and near-infrared region. Room lights and
sunlight will contaminate your spectra. Take care not to misinterpret room lights or
sunlight features as Raman features.
Sampling Liquids
Liquid samples can be analyzed by immersing the FOP in the liquid of inte r est or by analyzing
through the container.
Immersion
The FOP can only be immersed in liquid if you have purchased an immersible fiber probe
sleeve. Do not totally submerge the probe. It is only acceptable to immerse the probe up to
1 inch from the top of the sleeve. It may be useful to focus the sample by sliding the FOP
inside the immersion sleeve to obtain the best spectrum. Note that immersion sleeves
contain either quartz or sapphire windows. These windows can yield a detailed Raman
spectrum – do not confuse this with the spectrum of the sample.
62 . RamanMicro 200 Series Getting Started Guide
The standard FOP is not designed for immersion use
Immersion sleeve
Fiber optic probe
Figure 29 Fiber optic probe with the immersion sleeve detached
Non Immersion
When using 785 nm laser s, many types of glass exhibit fluorescent bands between 1400 –
1600 cm
ensure that glass-derived fluorescence bands do not pose a problem. With 785 nm
excitation, fused silica (quartz) should be used instead of glass where possible as this does
not yield fluorescent peaks. Pyrex is also excellent for Raman analysis. When working
through containers it is best to try different probe positions. If the liquid of interest is not
absorbing it may be best to focus through the container by placing the FOP in contact wit h
the container.
If this glass fluorescence is a problem and it is not possible to use quartz containers, then it
is possible to acquire a spectrum of the container, and spectrally subtract this from a ny
spectra containing this fluorescent signature.
Sampling Slurries/Highly absorbing species
Please refer to
within the sleeve can result in a dramatic improvement in spectral quality.
−1
. Glass spectra of the container should be obtaine d before analyzing t he sample to
Sampling Liquids
. For sample immersion care taken in focusing the probe
Sampling Gases
Although Raman scattering from gas samples is very weak, the FOP can be used to measure
gas phase under long acquisition times.
sealed against minor splashes. However, immer sion in solvents will
cause serious damage. Ensure an immersion sleeve is in place before
WARNING
immersing your FOP; see
have any questions in relation to sampling using the FOP.
. The FOP is
Figure 29. Please contact PerkinElmer if you
Using the RamanMi cro 200 Series . 63
General SOPs
The RamanMicro 200 combined with the Spectrum software is a very flexible instrument.
This chapter demonstrates how to perform a number of tasks. The procedures provided may
not be the most straightforward or simple for the task you wish to perform, so treat these
Standard Operating Procedures (SOPs) as a starting point and with time you may wish to
develop your own SOPs which better suit your specific mode of use. These procedures also
act as a “learn by doing” section allowing you to explore the functionality of the software and
spectrometer.
The General SOPs demonstrate how to collect data when your samples are on the
microscope stage. For SOPs that demonstrate how to coll ect data using a fiber optic probe ,
please refer to
calibrating the spectrometer, see
NOTE: The RamanMicro 200 system can be purchased with either a manual stage or with a
motorized stage.
Fiber Optic Probe SOPs
SOP-1: Turning the Instrument On
1. Switch on the PC and monitor.
on page 76. For step-by-step i nstructions for
Calibration and Performance Verification SOPs
on page 78.
2. Login to Windows.
3. Ensure the laser power key on the front panel of the spectrometer is in the ARM
position.
4. Ensure the power supply is connected to a mains outlet and that the outlet is switched
on.
5. To turn on the spectrometer, press the PSU power switch ON (Figure 30):
- ON Ο - OFF
Figure 30 The PSU power switch
64 . RamanMicro 200 Series Getting Started Guide
6. Check that the Power LED on the front panel is On.
The Laser LED and the Active LED should be off. If the Power LED is not on, please turn
the instrument off at the PSU and check all of the connections, and any power socket
switches and then switch the PSU on again.
7. Switch on the microscope and the microscope illumination.
8. If you have a motorized microscope stage, switch on the stage controller box.
9. From the Start menu select All Programs; the PerkinElmer Applications group; the
Spectrum sub-group and then the Spectrum application.
The Spectrum start-up screen is displayed; followed by a dialog that may require your
login details:
10. If required, enter your User name and Password, and then click OK.
An instrument selection dialog is displayed.
11. Select the Instrument you want to use.
OR
If you want to work with data that has been collected previously, without connecting to
an instrument, select Work Offline.
12. Click OK.
Spectrum starts.
If you selected to connect to the instrument, it will prepare for use by turning on the
laser and cooling the detector to its operating temperature (usually −50 °C). Messages
will be displayed in the Status bar at the bottom of the workspace, and in the Scan
icons on the Measurement bar.
NOTE: The CCD detector will become more stable the longer it is cooled.
If you have a motorized XYZ stage and have just switched on the stage controller box
you will be required to reset the stage before you can use the stage.
To adjust the height of the stage and set the limits of the stage travel, refer to SOP-3.
Using the RamanMi cro 200 Series . 65
SOP-2: Turning the Instrument Off
1. Shut down Spectrum software by selecting Exit from the File menu to return to the
Windows operating environment.
2. Switch off the PC and monitor.
3. Switch off the spectrometer by pressing the power switches on the PSUs to OFF:
- ON
4. Switch off the microscope illumination, and then the microscope.
5. If appropriate, switch off the stage controller box.
However, we recommend that you leave the stage controller box switched on, even if
you switch off the instrument and mi croscope. If the stage control box is switched off,
you will need to reset the stage upon restarting the software.
6. If appropriate, ensure that all sample holders have been removed from the stage in
preparation for the next time the instrument is used.
After Spectrum closes, if you do not switch off the instrument it will remain active for
5 minutes. This is so that you do not have to wait for the CCD detector to cool and the laser
to stabilize should you need to exit Spectrum and then restart the software immediately, or
when changing methods in the AssureID software a pplication.
NOTE: For information on how to use AssureID with a triggered fiber optic probe, refer to
the AssureID on-scr een help.
The instrument will shut down automatically after the appropriate time period. Yo u can shut
down the instrument by right clicking on the Raman instrument icon in the Notification area
of the Windows Taskbar and accessing the shortcut menu.
Ο - OFF
You can shut down your PC when the Raman instrument icon is displayed. The system will
shut down.
NOTE: If Spectrum software stops responding, refer to
for details of how to shut down the instrument and restart the software.
Troubleshooting
starting on page 94
66 . RamanMicro 200 Series Getting Started Guide
Position of
SOP-3: Setting the Limits of the Motorized Stage
NOTE: If you move the stage, you may also have to check the alignment of the microscope
objectives. Refer to
You may need to adjust the height o f the stage, for example, when you want to measure
thicker samples.
After moving the stage, you will need to reset the limits in the software.
CAUTION
1. Ensure the instrument is switched on by following SOP-1.
If the stage controller box was switched off, you will be required to reset the stage
before using the motorized stage.
2. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
Aligning the Microscope Objectives
on page 50.
3. Select the Setup Instrument Basic tab.
4. Ensure that Microscope XYZ Stage is selected in the Accessory drop-down list.
The options availab le will depend upon your instrument configuration.
5. Ensure that the correct Microscope Objective is selected in the software.
6. Ensure that the pre -focusing leve r on the coarse adjustment control is on the off
position.
The pre-focusing lever is restricts the travel of the coarse adjustment control, and is
located on the left-hand side of the microscope. The lever is in the off position when it
is rotated counter-clockwise away from you when facing the microscope. Refer to the
Olympus manual provided with your microscope for further information.
7. Loosen the fixing screw on the side of the microscope (Figure 31).
fixing screw
Figure 31 Adjusting the heig ht of the motorized stage
8. Manually move the stage down until the stage is at its lowest position.
Using the RamanMi cro 200 Series . 67
9. Re-secure the fixing screw to fix the height of the stage.
10. Select the Setup Microscope XYZ tab.
Figure 32 Setup Microscope XYZ Stage tab showing the Live Video view
11. Select Toolbox and then select Reset Stag e from the sub-menu.
12. Follow the prompts on-screen.
During the Reset Stage procedure, you will be prompted to use the joystick to move the
stage to the point of lowest travel. If you place yo ur hand carefully on the coarse
adjustment control, you will feel it move (as it starts to counter-rotate with the fine
focus control) when you have reached the lower limit. Stop using the joystick at this
point.
13. When the stage ha s been reset, use the joystick or the stage position controls in the
software to move the microscope stage up to 0 mm (approximately 25 mm travel).
The stage will stop at some point between 0 mm and 1 mm, at the maximum length of
travel for the z-axis motor.
14. Ensure that you are now at the maximum point of travel for the stage by turning the
coarse adjustment control.
It may move slightly, but should be nearly at the maximum.
15. Place your sample on the stage.
Using a hexagonal key, loosen the fixing screw on the side of the microscope
(Figure 31).
16. Carefully, move the stage up manually until it is at the highest position possible without
the sample touching the objective.
Be very careful not to touch the objective.
CAUTION
The performance of the Raman microscope will be affected by dirt on
the surface of, or damage to, the objectives.
NOTE: When using long working-distance objectives (for example, Long Working Distance
Objective Lens K it, L1323347), the working distance depends on the objective
(usually, 2–10 mm).
68 . RamanMicro 200 Series Getting Started Guide
se
when the stage has reached the lower or upper limits of travel. To see if
17. Re-secure the hexagonal fixing screw to fix the height of the stage.
18. Select Toolbox and then select Reset Stag e from the sub-menu.
19. Follow the prompts on-screen, as before.
You will need to repeat this process every time you adjust or install the stage.
Once you have reset the stage you should use the joystick ONLY to rai
or lower the stage. DO NOT try to adjust the stage manually, or you will
have to reset the stage.
CAUTION
When using the joystick, take care not to continue using the motors
you have moved outside the limits, che ck the z-axis focus dials on the
right-hand side of the microscope. If the two wheels counter-rotate, you
will need to reset the stage. If you continue to move the stage, the
cable can become trapped and the motor could be damaged.
SOP-4: Sample Positioning using a Manual Stage
1. Lower the microscope sample stage.
2. Place the sample on a microscope slide and place it on the stage.
Place your
sample here
Figure 33 Positioni ng the sample under the objectiv e lens
3. Select the 5x objective lens on the microscope.
NOTE: Do not touch the glass of the objective lenses when you select the appropriate
magnification. Do not push on an objective to rotate the nosep iece, but use the
revolving plastic ring.
4. Move the stage so that the sample is approximately lined up below the objective lens.
5. Select the Bright Fi eld (BF) switch position on the microscope.
6. Switch on the illumi nation source.
The switch is located on the right side of the microscope frame.
7. Set the required light intensity using the adjustment knob on the right side of the
microscope frame base.
Using the Rama nMicr o 20 0 Ser ie s . 69
8. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
9. Select the Setup Instrument Basic tab.
Figure 34 Setup Instrument Basic tab with the Microscope accessory selected
10. Ensure that Microscope is selected in the Accessory drop-down list.
The options available will depend upon your instrument configuration.
11. Select 5x from the Microscope Objective drop-down list.
The microscope is supplied with 5x, 20x and 50x objective lenses.
12. Select the appropriate Sample Holder from the drop-down list.
13. Select the Setup Microscope tab.
Figure 35 Setup Microscope tab
14. Focus on the sample using the micro scope z-axis control knobs.
Take care not to crush the sample or dirty the objectives. The working distances of the
objective lenses supplied as standard are 20 mm (5x), 1.3 mm (20x) and 0.38 mm
(50x).
15. If higher magnification is required, select the next most powerful objective lens on the
microscope.
16. Select the appropriate objective lens from the Microscope Objective drop-down list.
70 . RamanMicro 200 Series Getting Started Guide
17. Focus on the sample and again move the sample in the x and y directions, if necessary.
The crosshairs indicate the spot where analysis takes place.
NOTE: If there is an offset between the laser position and the cross hairs, use the Align
Video with Laser tool to adjust this. See the Spectrum on-screen help and
the Video Camera with the Laser Spot
18. When the sample is correctly positioned, switch off the illumination source.
19. Select the Dark Fie ld (DF) switch position on the microscope.
20. If the microscope light-proof enclosure is fitted, close the doors.
21. Click .
The RamanMicro 200 will start collecting data in the Live disp lay.
You can use Monitor to help you optimize the Exposure time and Laser power for your
sample, and adjust the focus of your sample to maximize the Raman signal. The data
displayed on-screen when monitoring cannot be saved. To save data you must collect
spectra.
on page 52 for more information.
Aligning
Click to exit the Live display.
You are now ready to acquire data. For more information see SOP-6.
The most appropriate microscope objective for your analyses will depend upon your samples.
High-magnification objective lenses give the most efficient Raman collection from thin
samples, such as films or wafers, but low-magnification objectives are better for macro
samples such as tablets, bulk plastics and tissue samples.
NOTE: Raman spectra are measured in the visible and near-infrared spectrum. Room
lighting and sunlight will contaminate your spectra. Take care not to misinterpret
room lighting or sunlight features as sample features.
SOP-5: Sample Positioning using a Motorized Sample Stage
When you are using a motorized stage, do not place your fingers
between the moving and fixed parts of the stage. The motors driving
the stage are powerful and do not stall easily.
WARNING
1. Select the 5x objective lens on the microscope.
2. Select the Bright Fi eld (BF) switch position on the microscope.
3. Switch on the illum ination source.
4. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
Using the RamanMi cro 200 Series . 71
Place your sample
5. Select the Setup Instrument Basic tab.
Figure 36 Setup Instrument Basic tab with the Microscope XYZ Stage accessory
selected
6. Ensure that Microscope XYZ Stage is selected in the Accessory drop-down list.
The options available will depend upon your instrument configuration.
7. Select 5x from the Microscope Objective drop-down list.
The microscope is supplied with 5x, 20x and 50x objective lenses.
8. Load your sample into a suitable sample holder.
9. Select the appropriate Sample Holder in the drop-down list, and follow the instructions
on-screen to load the sample holder onto the sample stage.
holder here
Figure 37 Sample position on motorized stage
72 . RamanMicro 200 Series Getting Started Guide
10. Select the Setup Microscope XYZ tab, and then select the Live Video view.
Figure 38 Setup Microscope XYZ Stage tab showing the Live Video view
11. Use the joystic k, or click Stage Position and then use the controls, to move the stage
so that the sample is approximately lined up below the objective lens and then focus on
the sample.
The crosshairs indicate the spot where analysis takes place.
The joystick operat es in the following manner:
• Left / Right joystick movement gives x movement of the stage.
• Top / Bottom joystick movement gives y movement of the stage.
• Twisting the joystick gives z movement of the stage (if purchased):
Left twist (counter clockwise) gives upward stage movement;
Right twist (clockwise) gives downward stage movement.
NOTE: You can also click Video Autofocus to perform an automatic video focus when
you are close to the sample surface (when the microsc ope illumination is on and the
switch position set to Bright Field). Alternatively, select the Dark Field switch position
on the microscope, and click Raman AutoFocus to focus using t he strongest
Raman signal.
12. If higher magnification is required, select the next most powerful objective lens on the
microscope.
The microscope is supplied with 5x, 20x and 50x objective lenses.
NOTE: Do not touch the glass of the objective lenses when you select the appropriate
magnification. Do not push on an objective to rotate the nosep iece, but use the
revolving plastic ring.
13. Select the appropriate objective lens from the Microscope Objective drop-down list.
x
14. Focus on the sample and, i f necessary, move the stage in
accurately position the sample.
15. When the sample is correctly positioned, switch off the illumina tion source.
16. Select the Dark Fie ld (DF) switch position on the microscope.
and y directions to
Using the RamanMi cro 200 Series . 73
17. If the microscope light-proof enclosure is fitted, close the doors.
18. Click .
The RamanMicro 200 will start collecting data in the Live display.
You can use Monitor to help you optimize the Exposure time and Laser power for your
sample. The data displayed on-screen when monitoring cannot be saved. To save data
you must collect spectra.
19. Adjust the Laser power (%) in the Live display to an appropriate value for your sample.
20. Enter an Exposure time (s).
Changing the exposure time will automatically acquire another background.
21. Click to exit the Live display.
You are ready to acquire data. For more information see SOP-6.
The most appropriate objective for your analyses will depend upon your sample . High-
magnification objective lenses give the most efficient Raman collection from thin sa mples,
such as films or wafers, but low-magnification objectives are better for macro samples such
as tablets, bulk plastics and tissue samples.
NOTE: Raman spectra are measured in the visible and near-infrared spectrum. Room
lighting and sunlight may contaminate your spectra. Take care not to misinterpret
room lighting or sunlight features as sample features.
SOP-6: Setting-Up and Saving Experimental Parameters
This SOP is designed to act as a quick guide to help yo u start collecting data. The exact
setup for optimized spectral collection is sample dependent, and we recommend that you
modify these settings to suit your needs.
1. Start the Spectrum software and select Raman Instrument from the Set up menu.
The Setup Instrument tabs are displayed in the Dialog pane. They enable you to choose
the accessory, and setup and save the experimental parameters.
2. Select the Setup Instrument Basic tab.
Figure 39 Setup Instrument Basic tab
74 . RamanMicro 200 Series Getting Started Guide
3. Select Microscope or Microscope XYZ Stage as appropriate from the Accessory
drop-down list.
Depending on the Accessory you have selected, further options may become available.
4. Ensure that the appropriate Microscope Objective and Sample Holder are selected.
5. Using the Instrument Settings toolbar, check and set the instrument parameters, such
as the Start and End points of the scan range (in Raman Shift, for example, 3200 cm
to 100 cm
6. If required, adjust the Data Interval in Raman Shift (cm
The default data interval is 2 cm
-1
), and enter a unique Sample ID and Description.
−1
).
−1
. The Data Interval can be set on the Setup
Instrument Basic tab.
7. Set the Exposure Time and number of Accumulations.
You can define the Accumulations as a number of Exposures, or as a number of minutes
or seconds.
Typical values for Exposure Time and Accumualtions are 5 seconds and 5 exposures,
respectively. Spectra may be optimized by increasing or decreasing the Exposure Time
and number of exposures. To obtain a good signal-to-noise ratio, you should co-add a
number of spectra. This can be done by choosing a suitable number of exposures. A
miniumum of two exposures is recommended for cosmic ray removal.
8. If you wish to edit the data collection parameters, such as how background scans are
collected, select the Setup Instrument Data Collection tab.
To display the Setup Instrument tabs, select Raman Instrument from the Setup
menu. The tabs are displayed in the dialog pane.
9. In the Background Exposures section, we recommend that you select As samples.
This sets the number of background scans to the same value as the number of sample
scans. You may also choose one of the Background Collection options to define when a
background will be taken.
-1
10. If required, select any Auto Processing options.
These options, if selected, will be applied to all spectra collected, and cannot be
undone.
11. If you select Save after each measurement, your spectra will automatically be saved
using the Sample ID.
12. If you have a motorized stage, select the Setup Microscope XYZ Stage tab, and then
select the Sampling Pattern.
Select the sampling pattern type from Single Point, Super Macro Point, Custom Grid
and Custom Line. Any spectra collect ed using SuperMacro, Custom Grid or Custom
Line sampling patterns are coadded to produce one spectrum.
For more information, see the Spectrum on-screen Help.
13. If you have a motorize d stage, select any Focus options.
You can select to perform a Raman AutoFocus or Video AutoFocus before each
sample measurement.
14. When you have finished defining your experimental param eters, position and optimize
conditions for your sample as described in SOP-4 and SOP-5.
15. Click .
Using the RamanMi cro 200 Series . 75
Spectra are always background corrected. If there is not a valid background in memory, a
background will be acquired at this point. You will see a progress bar for the background
collection on the Live tab. You can enter the number of backgrounds on the Setup
Instrument Data Collection. When the collection of backgrounds is complete, the software
will automatically begin collecting spectra. The upper progress bar shows the progress of the
current scan (%), while the lower progress bar shows the overall progre ss of the
measurement (for example, 1 of 2 scans) if, for example, you have set up Repeat collections.
If you exit Spectrum software the instrument settings will be remembered when you restart
Spectrum, providing that they have been used to collect data. If you want to return to the
default instrument settings, click Restore Defaults on the Setup Instrument Basic tab. If you
wish these parameters to be stored as a new set of instrument settings, select Load and
Save on the Setup Inst rument Basic tab.
NOTE: If you have a motorized stage, for information on how use the Setup Microscope XYZ
Stage tab to set up Markers, Cell Markers, Maps and Line Scans, and to collect a
Visible Image Survey, refer to the Spectrum on-screen help.
NOTE: If you are using a multi-well plate on the motorized stage on the microscope, then
you must insert the multi-well plate holder (L1321675) onto the stage before adding
the plate.
76 . RamanMicro 200 Series Getting Started Guide
Fiber Optic Probe SOPs
NOTE: The SOPs included on the following pages are only appropriate for a standard fiber
optic probe. For details of how to acquire data using a triggered fiber optic probe, see
the
Raman Triggered Fiber Optic Probe
SOP-7: Selecting the Fiber Optic Probe
1. Ensure that the RamanMicro 200 is switched on and that the fiber optic probe is
connected to the instrument.
If not follow SOP-1.
2. Select Raman Instrument from the Setup menu in Spectrum.
The Setup Instrument tabs are displayed in the Dialog pane.
3. Select the Setup Instrument Basic tab.
leaflet (L1321940).
Figure 40 Setup Instrument Basic tab
4. Select the Fiber Optic Probe you want to use from the Accessory drop-down list.
The following warning message is displayed.
Using the RamanMi cro 200 Series . 77
5. Ensure that laser safety precautions are being observed, and then click Continue.
If you have any doubt s about the required safety precautions click Cancel.
When the laser is on and the shutter is open the Laser Status icon on the Measurement
bar will be updated to Class 3B Laser Status . Laser light will be emitted from the
probe during data collection and when the software is Monitoring.
The probe can now be positioned so as to achieve the best spectrum. See SOP-8.
SOP-8: Sample Positioning when using a Fiber Optic Probe
Monitor or Preview modes allow you to collect spectral data while optimizing the probe
position and the laser power and exposure time, for example, to enhance the signal-to-noise
ratio of spectral data. When measuring samples inside a container (for example, a bottle) it
can be used to ensure that you are focused on the material of interest and no t the container.
The data that is displ ayed on the Live tab when monit oring cannot be saved.
NOTE: To see the Live display, the Show live display option must be selected on the Setup
Instrument Data Collecti o n ta b.
1. Click .
The Live tab will be displayed and real-time spectral acquisition will commence.
2. Adjust the Laser power (%) on the Live tab to an appropriate value for your sample.
3. Adjust the Exposure time (s).
This will automatically acquire another background.
The sample may now be positioned and aligned correctly for optimized data collection.
The spectral data can be optimized by adjusting the distance between the fiber optic
probe and the sample.
NOTE: When positioning your sample, you should ensure that you are not collecting data
from any substrate used to support your sample. It is advisable to have 1–2 cm of
free space behind the sample. Alternatively, you can subtract the spectrum of your
substrate from your sample spectrum.
4. Click to exit the Live display.
You can now collect and save spectral data. To set up an experiment and collect data using
SOP-6
the FOP please refer to
NOTE: If you have Show Live Display selected on the Setup Instrument Data Collection tab
and you select Preview on the Sample Table, then you can preview each sample in
the Live display after pressing Scan. When you ha ve a djuste d the se tt ings, cl ic k
Scan again to collect data.
.
78 . RamanMicro 200 Series Getting Started Guide
Calibration and Performance Verification SOPs
Several different automated calibration tests ar e included in the software:
• Wavelength (x-axis) calibration uses the position of precisely known lines in a neon
emission spectrum to calibrate the det ector.
• Intensity (y-axis) calibration is performed using coated glass with a well characterized
emission spectrum. This is used to correct for non-linearity in detector response.
• Laser frequency determines the exact laser wavelength.
An additional wavelength calibration verification routine has been included in the soft ware,
which compares a series of peaks for polystyrene ( ASTM E 1840) with values collected using
a polystyrene sample. These calibration/verification tests allow for in-house instrument
performance verification.
Intensity calibration is only performed if the instrument has been moved, or as part of an
annual service, or following a service repair of the instrument or accessory, or if there have
been appreciable temperature variations in the local environment.
Backgrounds are always collected during a calibration procedure. The background measures
the bias and dark signal over the selected area of the CCD. This is a necessary part of the
calibration routines, so it is always performed.
The spectrometer has no moving parts in its detection system; therefore frequent cali bra ti on
is not necessary.
We recommend that you calibrate the instrument in accordance with the following SOPs.
SOP-9: Calibration/Performance Verification
We recommend that wavelength calibration verification is performed on a regular basis to
ensure that the instrument is performing within sp ecification. This can be once a day or once
a week depending on your requirements and the laboratory environment.
When carrying out the calibratio n/verification procedures, the instrument laser power should
be set to 100% and the Data Interval to 4 cm
On completion of the wavelength calibration verification, the results of the verification (or
calibration) procedure is displayed. A spectrum detailing the outcome of the verification
procedure is saved to the folder …\pel_data\config\Raman\Calibration Files\Logs with the
filename Calibration Report [Name of calibration][Date].sp
IMPORTANT: If the instrument passes the wavelength calibration verification procedure,
the instrument is optimized for performance and does not require wavelength calibration.
−1
or smaller.
Wavelength Calibration
Laser Frequency
Calibration
focus of polystyrene
Completed
Pass
Using the RamanMi cro 200 Series . 79
Verification
Check Laser Key is
turned on and laser
power is set to
100%
Wavelength
Calibration
Fail
Peak
wavelengths
outside limits
Fail
Intensity
low
Finished.
Instrument ready
for use
Check position and
sample
Figure 41 System verification / calibration flow diagram
If wavelength calibration verification fails and t he Description of the calibration report
spectrum states that peak wavelengths are outside limits, then wavelength and laser
frequency calibrations should be performed. When these calibrations ha ve been completed
successfully, repeat the wavelength verification to ensure that the instrument has been
brought back within specification.
If the wavelength calibration verification fails because the spectral intensity is low, then this
may also be due to the calibration sample holder not being placed in the sample accessory,
or that it is not correctly positioned, optimally focused or because the laser key is not
switched on, or the laser power is too low. Then repeat the wavelength calibration
verification to ensure that instrument has been br ought back within specification. If the
instrument fails the wavelength calibration verification again, please contact PerkinElmer.
NOTE: Stray light could cause the wavelength calibration verification to fail. If a test fails,
particularly if you are using the microscope without an enclosure, en sure that no light
is entering the Raman system.
SOP-10: Wavelength Calibration Verification
To perform the Wavelength Calibration Verification with the Microscope 50x objective
selected:
1. Ensure that the RamanMicro 200 is switched on.
If not follow SOP-1.
2. Select the 50x objective lens on the microscope.
3. Place a polystyrene sample (L1181228) in a suitable sample holder.
80 . RamanMicro 200 Series Getting Started Guide
4. Select Raman Instrument from the Setup menu.
The Setup Instrument t abs are displayed.
5. On the Setup Instrument Basic tab s elect Microscope or Microscope XYZ Stage as
appropriate from the Accessory drop-down list
6. Select Microscope 50x from the Objective drop-down list.
7. Select the appropriate Sample Holder in the drop-down list and load the sample holder
onto the sample stage, following the instructions on-screen if you have a motorized
stage.
8. Select the Setup Instrument Advanced tab and ensure that the Laser Power is set to
100%.
9. Select the Dark Field (DF) switch position on the microscope and switch off the
illumination.
10. Cover the microscope with a dark cloth or, if fitted, close the light-proof enclosure doors
and ensure that caps are placed on any external fiber optic probes or accessories, to
minimize stray light from interfering with the calibration verification.
11. Focus on the polystyrene sample.
If you have a motorized stage you can use the joystick or Stage Position controls on the
SetupMicroscope XYZ Stage tab. Alternatively, click Raman AutoFocus to focus
using the strongest Raman signal. The Focus Limits should be set to Full Range.
If you do not have a motorized stage, you may find it useful to Monitor your sample to
view the Raman signal while manually adjusting the focus.
12. When the polystyrene sample is optimally focused for Raman signal, select Toolbox on
the Setup Instrument Advanced tab.
The Adjustments Toolbox dialog is displayed.
13. Select .
The Wavelength Calibration Verification dialog is disp layed.
14. Ensure that you have followed the instructions on the dialog, and then click Next.
The wavelength calibration verification routine beg ins. This will take a few minutes.
When the verification is complete, the result is displayed.
15. Remove the polystyrene sample and store safely.
The instrument is now ready for use.
Using the RamanMi cro 200 Series . 81
In the unlikely event that the wavel engt h calibration verification should fail, follow Figure 41.
If the wavelength calibration verification fails for the microscope, this does not necessarily
mean that the performance of any other accessories attached to your instrument is also
affected.
SOP-11: Wavelength Calibration
Wavelength calibration should only be performed if the instrument has failed a wavelength
calibration verification.
1. Ensure that the RamanMicro 200 is switched on.
If not, follow SOP-1.
2. Attach the external neon calibration accessory (L1320220) to the rear panel of the
spectrometer module.
Push the large connector into the socket labeled CALIBRATION or CAL ACCY and
carefully attach the small cable to the COL coupler below the LED FIBER 1 connector.
Figure 42 Attaching the calibration accessory
NOTE: As wavelength calibration is not required on a re g ula r basis, the external neon
calibration accessory is not included in the basic RamanMicro 200 package. The
external neon calibration accessory is included in the RamanMicro Calibration Kit
(L1320266), whi ch can be purcha se d se pa ra te ly.
3. Rotate the objective nosepiece until mid-way between two objectives.
4. Cover the microscope with a dark cloth or, if fitted, close the light-proof enclosure doors
and ensure that caps are placed on any external fiber optic probes or accessories, to
minimize stray light from interfering with the calibration.
5. Select the Dark Field (DF) switch position on the microscope and switch off the
illumination.
6. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
7. Select the Setup Instrument Advanced tab and click Toolbox.
The Adjustments Toolbox dialog is displayed.
8. Select .
The Wavelength Calibration dialog is displayed.
82 . RamanMicro 200 Series Getting Started Guide
9. Ensure that you ha ve followed the i nstructions on the dialog, and then click Next.
The instrument will now perform the wavelength calibration routine. This will take some
time. When the calibration is complete, the result is displayed.
10. Disconnect the neon calibration accessory and refit the screw-in cap to the port.
The instrument is now wavelength cali brated.
SOP-12: Laser Frequency Calibration
Laser frequency calibration should only be performed if the instrume nt has failed a
wavelength calibration verification.
To calibrate the microscope with the Microscope 50x objec tive selected:
1. Ensure that the RamanMicro 200 is switched on.
If not follow SOP-1.
2. Select the 50x objective lens on the microscope.
3. Place a polystyrene sample (L1181228) in a suitable sample holder.
4. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
5. On the Setup Instrument Basic tab select Microscope or Microscope XYZ Stage as
appropriate from the Accessory drop-down list
6. Select Microscope 50x from the Objective drop-down list.
7. Select the appropriate Sample Holder in the drop-down list and load the sample holder
onto the sample stage, following the instructions on-screen if you have a motorized
stage.
8. Select the Dark Field (DF) switch position on the microscope and switch off the
illumination.
9. Cover the microscope with a dark cloth or, if fitted, close the light-proof enclosure doors
and ensure that caps are placed on any external fiber optic probes or accessories, to
minimize stray light from interfering with the calibration.
10. Select the Setup Instrument Advanced tab and ensure that the Laser Power is set to
100%.
Using the RamanMi cro 200 Series . 83
11. Focus on the sample of polystyrene.
If you have a motorized stage you can use the joystick or St age Position controls on the
SetupMicroscope XYZ Stage tab. Alternatively, click Raman AutoFocus to focus
using the strongest Raman signal.
If you do not have a motorized stage, you may find it useful to Monitor your sample to
view the Raman signal while manually adjusting the focus.
12. When the polystyrene sample is optimally foc used for Raman signal, select the Setup
Instrument Advanced tab and click Toolbox.
The Adjustments Toolbox dialog is displayed.
13. Select .
The Frequency Calibration dialog is displayed.
14. Ensure that you have followed the instructions on the dialog, and then click Next.
The instrument will now perform the laser frequency calibration routine. This will take a
few minutes. When the calibration is complete, the result is displayed.
The instrument in now laser frequency calibrated.
In the unlikely event that the laser frequency calibration routine should fail, please perform a
wavelength calibration and repeat the laser frequency calibration.
NOTE: If the calibration fails again, please contact your PerkinElmer Service Representative.
SOP-13: Intensity Calibration
The intensity calibration must be performed using the National Institute of Standards and
Technology (NIS T) Standard Refer ence Material. NIST SRM 2241 is the relative intensity
correction standard for Raman spectroscopy when using 785 nm excitation.
NOTE: NIST calibration is temperature dependent. If you intend to perform quantitative
analyses, you should ensure that your instrument is NIST calibrated at the
temperature at which data is to be acquired. Otherwise, any data should be used for
qualitative purposes only.
The NIST intensity correction is valid in the recommended operating temperature
range 20 °C – 25 °C. If you move the instrument to an environment outside this
temperature range, that is 15 °C – 20 °C or 25 °C – 30 °C, we strongly recommend
that you perform a NIST intensity correction at this new temperature. Contact your
PerkinElmer Raman Business Unit.
84 . RamanMicro 200 Series Getting Started Guide
Intensity calibration should typically be performed on an annual basis – that is, during an
annual instrument service – but al so whenever the instrument is moved, following a service
repair or when there has been significant temperature variation in the local environment.
NOTE: As intensity calibration is not required on a regular basis, NIST Standards are not
included in the basic RamanMicro 200 package. Standard Reference Materials can be
purchased from PerkinElmer (L1321831). Your instr ument will be intensity calibrated
prior to shipping and during installation.
NOTE: The intensity calibration should be performed for each sampling accessory. In the
case of the RamanMicro 300 Accessory, an intensity calibration is required for each
objective lens.
1. Ensure that the RamanMicro 200 is switched on.
If not follow SOP-1.
2. Select the 50x objective lens on the microscope.
3. Place the NIST standard in a suitable sample holder.
4. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
5. Select Raman Instrument from the Setup menu.
The Setup Instrument tabs are displayed.
6. On the Setup Instrument Basic tab s elect Microscope or Microscope XYZ Stage as
appropriate from the Accessory drop-down list
7. Select Microscope 50x from the Objective drop-down list.
8. Select the appropriate Sample Holder in the drop-down list and load the sample holder
onto the sample stage, following the instru ct ions on-screen if you have a motorized
stage.
9. Select the Dark Field (DF) switch position on the microscope and switch off the
illumination.
10. Cover the microscope with a dark cloth or, if fitted, close the light-proof enclosure doors
and ensure that caps are placed on any external fiber optic probes or accessories, to
minimize stray light from interfering with the calibration.
11. Select the Setup Instrument Advanced tab, and ensure that the Laser Power is set to
100%.
12. Focus on the appropriate NIST standard with the dull surface facing upw ards.
If you have a motorized stage you can use the joystick or Stage Position controls on the
SetupMicroscope XYZ Stage tab. Alternatively, click Raman AutoFocus to focus
using the strongest Raman signal. The Focus Limits should be set to Full Range.
If you do not have a motorized stage, you may find it useful to Monitor your sample to
view the Raman signal while manually adjusting the focus.
13. When the polystyrene sample is optimally foc used for Raman signal, select Toolbox on
the Setup Instrument Advanced tab.
The Adjustments Toolbox dialog is displayed.
Using the RamanMi cro 200 Series . 85
14. Select .
The Intensity Calibration dialog is displayed.
15. Ensure that you have followed the instructions on the dialog, and then cl ick Next.
The instrument will now perform the intensity calibration routine. This will take some
time. When the calibration is complete, the result is displayed.
Any data acquired will now be intensity corrected.
In the unlikely event that the intensity calibration routine should fail, please ensure that
the NIST sample is correctly positioned, and that the laser key switch is set to on
(ARM).
16. Repeat steps 2–15 for each objective lens.
17. Remove the intensity correction standard and store safely.
Summary
• The only calibration procedure that needs to be performed on a regular basis is the
wavelength calibration verification procedure.
• The wavelength and laser frequency calibrations should only be performed if the
instrument fails the wavelength calibration verification procedure.
• We recommend that the intensity calibration is performed on a yearly basis, unless the
environment (temperature and/o r humidity) changes significantly, or the instrument is
moved or repaired.
86 . RamanMicro 200 Series Getting Started Guide
Additional Parts and Upgrades
Microscope Objective Lenses
Part Number Description
L1323336 100x objective lens
L1323347 Long working distance objective lenses upgrade kit
Contains 5x and 20 x long working distance objective lenses
L1323343 5x long working distance objective lens
L1323344 20x long working distance objective lens
L1323345 50x long working distance objective lens
L1323346 100x long working distance objective lens
Sampling Accessories
Part Number Description
L1323514 Quartz microscope slide, 25 × 75 × 1 mm
L1860335 Tablet holder kit for motorized stage
L1860336 Powder holder kit for motorized stage
L1321675 Multi-well plate holder for motorized stage
L1321676 Multi microscope slide holder
Holds 4 standard (25 × 75 × 1 mm) microscope slides
L1321677 Universal sample holder
Holds standard (25 × 75 × 1 mm) microscope slides,
50 × 75 mm glass slides, and Petri dishes up to 60 mm
L1860635/
L1860634
N1870184/
N1870188
Linkam temperature controlled stage (110–130 V AC/220–240 V AC)
Hot stage accessory (110 V A C/230 V AC)
For use with long w orking distance objectives
SERS Consumables
Part Number Description
Using the RamanMi cro 200 Series . 87
L1329300
L1329307
L1329308
L1329301 Aggregating agent – MgSO4 0.16 M in milli-Q water (20 ml)
L1329303 Standard crystal violet pigment (0.25 ml)
L1329304 Disposable micropipettes – 200 µl (100 pack)
L1329305 Disposable micropipettes – 50 µl (100 pack)
L1329306 Disposable micropipettes – 2 µl (100 pack)
SERS gold colloid – 50 nm particles (20 ml)
SERS gold colloid – 100 nm particles (20 ml)
SERS gold colloid – 200 nm particles (20 ml)
Safety and Calibration
Part Number Description
L1320262 Raman laser safety kit (785 nm)
Contains external interlock cable, laser safety goggles, laser detection card,
and door warning label
L1320266 RamanMicro calibration kit
L1321843 sCAL Raman calibration standard
L1323503 Replacement laser keys
L1323518 Laser Glasses
88 . RamanMicro 200 Series Getting Started Guide
Fiber Optic Probes
Part Number Description
L1320002
L1320003 Liquid immersion sleeve for Raman fiber optic probe
L1320011
L1320012
L1320013
L1320030 Triggered Fiber Optic Probe, 5 m cable
L1320031 Triggered Fiber Optic Probe, 20 m cable
L1320324 Raman probe fixed spacer kit (7.5 mm)
L1320325 Raman probe adjustable spacer kit
L1320070 Short immersion sleeve – for use with L13200 30 and L1320031
Raman fiber optic probe, 7.5 mm working distance, maximum working
temperature 80 °C, 5 M cabling
Raman probe – long, 350 mm long, suitable for non-c ontact and liquid
immersion. Maximum working temperature 200 °C, maximum working
pressure 1500 psi, 5 M cabling
Raman probe – max, suitable for non-contact and liquid immersion.
Maximum working temperature 500 °C, maximum working pressure
3000 psi, 5 M cabling
Purge Jacket for Raman probe – max, required to enable max probe
operation at temperatures between 200 °C and 500 °C
L1320071 Ultra-long working distance lens adaptor
L1320095
For information about customized working distance or cable length fiber optic probes, contact
your PerkinElmer Sales Representative.
Fiber probe sample holder (for measurement of samples in small vials and
cuvettes)
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