PerkinElmer RamanMicro 300 Series User manual

RamanMicro 300 Series
Getting Started Guide
MOLECULAR SPECTROSCOPY

2 . RamanMicro 300 Getting Started Guide
Release History
Part Number Release Publication Date
L1321862 B May 2009
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: [e-mail]
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 © 2009 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 300 Getting Started Guide . 3
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
Environmental Conditions ............................................................................. 14
Electrical Safety........................................................................................... 15
Power Cords......................................................................................... 15
Laser Safety................................................................................................ 16
Class 3B Safety Precautions.................................................................... 17
General Laser Safety Issues.................................................................... 18
Warning Labels ........................................................................................... 20
Labels on the Exterior of the Microscope.................................................. 20
Raman History, Theory and Instrumentation..........................................21
The History of Raman Spectroscopy............................................................... 22
Raman Theory ............................................................................................ 23
The RamanMicro 300 ................................................................................... 25
The Echelle Spectrograph....................................................................... 27
Preparing Your Facility ............................................................................29
Specifications.............................................................................................. 30
PC Requirements......................................................................................... 31
Hardware Requirements......................................................................... 31
Software Requirements.......................................................................... 32
Windows Administrator Level.................................................................. 32
Finding a Suitable Location for the RamanMicro 300........................................ 33
Instrument Dimensions ................................................................................ 34
Installing Your RamanMicro 300.............................................................35
Unpacking the Instrument ............................................................................ 36
Lifting the Instrument .................................................................................. 38
Assembly of the RamanMicro 300.................................................................. 39
Fitting the Stage on the Microscope......................................................... 39
Connecting the Motorized Stage.............................................................. 40
Attaching the Illumination Source............................................................ 42
Connecting the Raman Module to the Microscope ..................................... 43
Connecting the Alignment Camera to the Raman Module........................... 43
Connecting the RamanMicro 300 to the Spectrometer Module .................... 44
Installing Spectrum Software ........................................................................ 46
Before you Start.................................................................................... 46
Installation Procedure............................................................................ 47
Alignment of the Microscope......................................................................... 63
Aligning the Microscope Objectives.......................................................... 63
Aligning the Raman Module.................................................................... 64
Moving the Instrument................................................................................. 67

4 . RamanMicro 300 Getting Started Guide
Using the RamanMicro 300......................................................................69
General SOPs.............................................................................................. 70
SOP-1: Turning the Raman System On .................................................... 70
SOP-2: Turning the Raman System Off .................................................... 71
SOP-3: Setting the Limits of the Motorized Stage ...................................... 71
SOP-4: Sample Positioning using a Manual Stage...................................... 73
SOP-5: Sample Positioning using a Motorized Stage .................................. 74
SOP-6: Setting-Up and Saving Experimental Parameters............................ 77
SOP-7: Defining Scan Parameters and Starting Data Collection................... 80
Calibration and Performance Verification SOPs ................................................ 81
SOP-8: Calibration/Performance Verification............................................. 81
SOP-9: Wavelength Calibration Verification of the RamanMicro 300............. 83
SOP-10: Wavelength Calibration of the RamanMicro 300............................ 84
SOP-11: Laser Frequency Calibration of the RamanMicro 300..................... 85
SOP-12: Intensity Calibration of the RamanMicro 300................................ 86
Summary.............................................................................................. 87
Additional Parts and Upgrades....................................................................... 88
Microscope Objective Lenses .................................................................. 88
Sampling Accessories............................................................................. 88
Safety and Calibration............................................................................ 89
Microscope Laser-Safe Enclosure............................................................. 89
Maintenance and Troubleshooting ..........................................................91
Maintenance ............................................................................................... 92
Cleaning the Spectrometer and Power Supply Units................................... 92
Maintenance and Service of the Microscope.............................................. 93
Replacing the Stage Controller Fuse............................................................... 94
Troubleshooting .......................................................................................... 95
Hardware............................................................................................. 95
Software .............................................................................................. 98
Decontamination ........................................................................................100
WEEE Instructions for PerkinElmer Products ..................................................101

Introduction

6 . RamanMicro 300 Getting Started Guide
About This Manual
This Getting Started Guide contains all of the information that you will need to setup your
RamanMicro 300 Raman microscope and start collecting spectra.
The RamanMicro 300 Raman microscope is used in conjunction with the RamanStation 400F
Raman spectrometer, or with the RamanFlex 400 Series Raman spectrometer. It cannot be
used in isolation.
NOTE: For a detailed description of the RamanStation or RamanFlex installation procedure,
see the Getting Started Guide supplied with the instrument.
This Getting Started Guide is divided into the following chapters:
Introduction
This chapter contains an overview of the manual, and the conventions and warnings used in
the manual.
Safety Practices
This chapter outlines the electrical and laser safety issues. We recommend that all users read
this chapter.
Raman History, Theory and Instrumentation
This chapter contains a brief overview of the history and theory of Raman spectroscopy.
Preparing your Facility
This chapter shows you how to position your RamanMicro 300, and what utilities are
required, for example electricity, temperature control.
Installing your RamanMicro 300
This chapter contains instructions about how to assemble your RamanMicro 300.
Using the RamanMicro 300
This chapter contains a number of Standard Operating Procedures (SOPs). These are a series
of procedures which describe how to perform certain tasks, for example how to collect a
spectrum, or how to calibrate the spectrometer.
Maintenance and Troubleshooting
This chapter contains solutions to problems you may encounter.

Introduction . 7
Conventions Used in this Manual
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 instrument means the RamanMicro 300 Raman Microscope.
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 300 Getting Started Guide
CAUTION
We use the term CAUTION to inform you about situations that could
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
einen Geräteschaden zu vermeiden.
Caution (Bemærk)
Dette betyder, at den nævnte vejledning skal overholdes nøje for at
undgå en beskadigelse af apparatet.
Caution (Advertencia)
Utilizamos el término CAUTION (ADVERTENCIA) para advertir sobre
situaciones que pueden provocar averías graves en este equipo o en
otros. En recuadros éste se proporciona información sobre este tipo de
circunstancias.
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
figurent dans un encadré semblable à celui-ci.
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.

Introduction . 9
WARNING
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.
Warning (Warnung)
Bedeutet, daß es bei Nichtbeachten der genannten Anweisung zu einer
Verletzung des Benutzers kommen kann.
Warning (Advarsel)
Betyder, at brugeren kan blive kvæstet, hvis anvisningen ikke
overholdes.
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'utilisateur
ou à d'autres personnes. Les détails sur ces circonstances sont données
dans un encadré semblable à celui-ci.
Warning (Pericolo)
Con il termine WARNING (PERICOLO) vengono segnalate situazioni
che potrebbero provocare incidenti alle persone. Troverete
informazioni su tali circostanze in un riquadro come questo.
Warning (Waarschuwing)
Betekent dat, wanneer de genoemde aanwijzing niet in acht wordt
genomen, dit kan leiden tot verwondingen van de gebruiker.
Warning (Aviso)
Significa que a não observância da instrução referida poderá causar um
ferimentoao usuário.

10 . RamanMicro 300 Getting Started Guide

Safety Practices

12 . RamanMicro 300 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 instructions, and make sure that you understand the procedures explained.
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 spectrometers 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 Institute’s Z136 series of laser safety
standards.

Safety Practices . 13
General Operating Conditions
The RamanMicro 300 microscope has been designed and tested in accordance with
PerkinElmer specifications and in accordance with the safety requirements of the
International Electrotechnical Commission (IEC). The RamanMicro 300 system meets the
requirements of EC directive 2006/95/EC (low voltage directive).
Only use the RamanMicro 300 microscope indoors and under the following conditions:
Temperature 15 °C to 30 °C
Relative Humidity 75% 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 hazard involved.
Whenever it is likely that the RamanMicro 300 is unsafe, make it inoperative. The
RamanMicro 300 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 300 Getting Started Guide
Environmental Conditions
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;
•Mains fluctuations not exceeding ±10% of the nominal voltage.
WARNING
If the equipment is used in a manner not specified herein, the protection
provided by the equipment may be impaired.
CAUTION
Ensure that there is at least 15 cm (6”) between any cooling vents and
adjacent equipment or walls.

Safety Practices . 15
Electrical Safety
Power Cords
The power cords supplied to connect the microscope to the power point (socket) should be
appropriate for the country listed as the shipping destination.
WARNING
If the power cord is not appropriate for your country it must be removed
from use immediately and replaced.
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.

16 . RamanMicro 300 Getting Started Guide
Laser Safety
The RamanMicro 300 Raman microscope is used in conjunction with the RamanStation 400F
Raman spectrometer, or with the RamanFlex 400 Series Raman spectrometer. As part of the
system the RamanMicro 300 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.
WARNING
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
damaged, turn the instrument off, secure it against any unauthorized
operation and contact PerkinElmer for advice before using the instrument
again.
The RamanMicro 300 emits a focused beam of laser radiation at 785 nm from the microscope
objective lens. This radiation is classified as Class 3B. The maximum laser power accessible
at the stage is 100 mW.
WARNING
Avoid direct exposure to the laser beam. Do not look at the laser light or
scattered laser light. Laser light at 785 nm is nearly invisible and only
appears dim to the eye.
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
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 . 17
Class 3B Safety Precautions
You should follow Class 3B laser safety precautions 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 ANSI Z136.1-2000 Standard. 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
published by the national authorities in your country.
The following 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
information as the basis 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 with 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 custody of a
nominated person. Keys must never be issued to unauthorized 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 connected 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.

18 . RamanMicro 300 Getting Started Guide
General Laser Safety Issues
Laser Keyswitch
There is a captive laser key control switch on the rear panel of the RamanStation, or the
front panel of the RamanFlex. This key is required to operate the laser. A laser safety officer
or laboratory manager should control 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 on page 20.
Microscope Safety
The laser should also be turned off when the system is not in use. AVOID DIRECT
EXPOSURE TO THE BEAM.
Figure 1 Schematic of microscope
Class 3B laser
radiation is emitted
through the
objective lens

Safety Practices . 19
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, 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);
•Use an interlock switch on the door to the laboratory, if required.
WARNING
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 the safe light path through
the system.
WARNING
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, ensure that protective
covers are in place when FOPs are not in use.

20 . RamanMicro 300 Getting Started Guide
Warning Labels
When this label is attached to an instrument it means refer to the
manual to find out the nature of the potential hazard and any actions
which have to be taken.
Labels on the Exterior of the Microscope
Figure 2 Labels on the microscope
LASER APERTURE
CAUTION –CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM
CAUTION –CLASS 3B LASER
RADIATION WHEN OPEN
AVOID EXPOSURE TO BEAM