LabRAM User Guide

LabRAM User Guide
Page 1
LabRAM
User Guide
An Introduction to the
Software and Hardware
v.01 11/2004
Page 2
Contents
Section 1 - Introduction to the LabRAM hardware and software
4
Introduction to the LabRAM hardware
Introduction to the Labspec software
The Hardware toolbar
White light illumination – image acquisition
5
8
9
11
Section 2 - The LabSpec software for data acquisition
Acquiring a spectrum
Real time spectrum adjustment
Spectrum accumulation
Simple
Multiwindow
CREST
Independent spectral windows
12
13
13
13
14
14
16
18
Acquiring mapped images and profiles
Introduction
Setting up a mapping/profiling experiment
20
20
21
Other acquisition functions
Cosmic ray (random spike) removal
Autosave
Autofocus
Autoexposure
Extended white light imaging
24
25
26
27
28
24
Section 3 - The LabSpec software for data analysis
Data analysis functions
Arithmetic
Baseline correction
Correction
Profile
Smoothing and filtering
Fourier transformation
Peak labelling and band fitting
Band integration
Object dimensions
Object generation
ASCII multi-file save
Custom units
29
30
30
30
31
32
33
33
34
36
37
38
38
39
Page 3
Contents
Section 3 - The LabSpec software for data analysis
(continued)
Analysing mapped images and profiles
Analysing with cursors
Analysing with models
Displaying a mapped image of band position, width, area, etc
40
41
43
Data display functions
View
Colours/Axes
3D Option
Scale
45
45
46
46
Other software functions
Multi
Axe + text
Copy and paste
XY(Z) stage display
Units
More objects
Page set up (for printing)
40
45
47
47
47
47
47
48
48
49
Section 4 – A summary of software icons
A summary of software icons
50
51
Section 5 – Maintenance and calibration procedures
Calibrating the LabRAM
Changing laser wavelength
Calibrating the XY stage movement
Calibrating the laser spot position
Installing LabSpec software for data analysis
56
57
59
60
61
63
Section 6 – Contact details
Contact details for Horiba Jobin Yvon
66
67
Page 4
Section 1
Introduction to the LabRAM Hardware and Software
Page 5
Introduction to the LabRAM Hardware
The following diagram of a LabRAM HR system shows the typical layout. The
arrangement of stems and accessories is identical for the smaller LabRAM 300
instrument. The instrument can be considered in four parts:
1.
Lasers – the HeNe (633nm) laser is internal, whilst other lasers are external,
mounted on an extended chassis at the back of the system.
2.
Microscope – sampling is carried out through a standard optical microscope.
3.
Spectrometer – dispersing the Raman signal into its constituent parts for
detection by detector (usually CCD, but other formats can be used).
4.
Optics – for coupling the lasers to the sample, and carrying the Raman signal
through to the spectrometer.
LASERS
(behind main
unit)
OPTICS
SPECTROMETER
(running underneath
top optics)
Stem 1
Stem 2
Stem 3
Stem 4
MICROSCOPE
(with sampling accessories,
such as XY stage, Z motor,
heating cooling stages etc)
For an explanation of the Stem
functions, see next page.
Page 6
Introduction to the LabRAM Hardware - continued
To provide the greatest flexibility and versatility, the LabRAMs include a number of
simple stems (or push-pull bars) to allow fast switching between different
functionalities.
Stem 1
Stem 2 Stem 3
Stem 4
Stem 1
Present on
LabRAM
Present on
LabRAM HR
"
"
Operates camera beamsplitter.
DOWN = camera
UP = Raman
Function
Stem 2
!
!
LabRAM 300 – operates switching mirror for point
mode (IN) and line scan (OUT).
LabRAM HR – operates switching mirror for vis-NIR
(IN) and UV-vis (OUT), or point mode (IN) and line scan
(OUT).
Stem 3
!
!
Operates switching mirror for microscope (IN) and fibre
entrance (OUT)
!
LabRAM 300 – operates grating turret (in the LabRAM
HR this is motorised).
IN = high resolultion (grating 1)
OUT = low resolution (grating 2)
LabRAM HR – operates switching mirror for two
detectors.
IN = side detector
OUT = top detector
Stem 4
!
" = always present
! = sometimes present
Page 7
Introduction to the LabRAM Hardware - continued
In conjunction with the software, the LabRAM control box provides an interface
between the user and the instrument. An explanation of the switches on the front
panel is given below.
Laser shutter
switch
Internal diode
for back
alignment
Line scan
facility
UP = ON
DOWN = OFF
UP = ON
DOWN = OFF
UP = Shutter
open, laser on
sample
DOWN = laser
blocked by
shutter
Detector
Main key control
for internal
633nm laser
(off position
shown)
LASER
OFF
DIODE
ON
Shutter switch
Camera power
UP =
microscope
DOWN = fibre
entrance
UP = video
camera on
DOWN = video
camera off
SHUTTER
MICRO
SCANNER
ON
CAMERA
ON
UP = shutter
controlled by
detector 1
DOWN = shutter
controlled by
detector 2
LASER
ON
SHUTTER 1
SHUTTER 2
LASER INDICATOR
INTERLOCK INDICATOR
Power indicators for laser and interlocks – on when lit
Power indicator – on when lit
For more information on the hardware, including optical layout, please see the
LabRAM User Manual.
Page 8
Introduction to the LabSpec Software
The LabSpec software provides a complete data acquisition and analysis package
for use with the LabRAM.
The LabSpec 4.XX folder on the desk top contains four icons:
Configuration Set
Up – password
protected.
Reload data files
into LabSpec in
event of software
crash.
Open the LabSpec
software
Uninstall
the
software from the
hard drive.
The main LabSpec screen can be divided up into a number of regions, as shown
below:
Menu Bar
Data analysis
Acquisition
toolbar
toolbar
Graphical
manipulation
Spectrum display
Hardware toolbar
Cursor values
In the rare event of a software crash, it is recommended that the software is exited,
and then double click on the Software Reset icon in the main LabSpec folder on the
desk top. Click OK, and then re-open the software with the LabSpec 4.XX icon.
Note that LabSpec can also be used for simple data analysis (no instrument control)
on other computers. See page 63 for details on how to install LabSpec to do this.
Page 9
The Hardware Toolbar
The hardware tool bar at the bottom of the screen controls settings such as the laser,
filter wheel, confocal hole, slit, spectrometer and gratings.
Displays the current laser
wavelength used – the
precise
value
of
this
wavelength can be amended
by adjusting the value in the
white box.
Select the laser wavelength
to be used from a pre-set
list.
(Has no affect.)
Select a neutral density filter
to be placed in front of the
laser beam, to prevent
sample burning and heating.
Close the confocal hole.
Type in the value for the
confocal hole and press
return.
Close the slit.
Type in the value for the slit
and press return.
Reinitialise the hole – sends
the hole to a known
reference point, and then
back
to
the
position
displayed in the box.
Open the confocal hole to its
maximum value.
Reinitialise the slit – sends
the slit to a known reference
point, and then back to the
position displayed in the
box.
Open the slit to its maximum
value.
Page 10
The Hardware Toolbar - continued
Send the spectromter to
Zero order.
Type in the central spectral
position for the spectrometer
and press return.
Reinitialise the spectrometer
– sends to Zero order, and
back
to
the
position
displayed in the box.
Send the spectrometer to
the reference diode positon.
Check box for autoexposure
Number
of
accumulations.
average
Accumulation time.
Select grating.
Select microscope objective.
File label – prefix a file name
to
each
accumulated
spectrum
Page 11
White Light Illumination – Image Acquisition
The LabRAM is equipped with standard white light illumination of the sample, by
reflection and/or transmission. A colour camera linked to the software allows the
sample to be visualised, and the image captured on the computer and saved.
1.
Put the camera beam splitter into
place.
Beamsplitter
down
DOWN = camera
UP = Raman
2.
Turn on the white light illumination.
3.
In the software, click on the video
icon to start the camera read out.
4.
To stop the continuous readout,
click on the STOP icon on the top
right hand side of the screen.
5.
Remember to turn off the white light
illumination, and take the camera
beam splitter out (=UP) before
starting a Raman measurement.
White
light on
To illuminate by transmission only, remove the white light fibre optic from the top left
port of the microscope:
Page 12
Section 2
The LabSpec Software for Data Acquisition
Page 13
Acquiring a spectrum
The LabSpec software provides the user with a number of methods for acquiring a
single spectrum.
Real Time Spectrum Adjustment
This method is primarily designed for a fast display of the spectrum on screen
corresponding to a single shot window. The update is continuous, providing a useful
way of adjusting the focus position to maximise the Raman signal, and quickly
monitoring the stability of the spectrum.
Each spectrum displayed replaces the previously displayed spectrum – there is no
averaging or accumulation of the spectra, and no extended coverage possible. To do
this you must use Spectrum Accumulation (see below).
1.
Ensure the central spectrograph
position has been correctly selected
(remember, the position typed into
the “spectro.” window corresponds
to the central position of the
resulting spectrum).
2.
Choose
required
3.
Click on the Real Time Spectrum
Adjustment icon.
4.
To stop the continuous readout,
click on the STOP icon on the top
right hand side of the screen.
the
integration
time
Spectrum Accumulation
This method allows spectra to be acquired with multiple accumulations and averaging,
and/or with coverage over extended regions.
Click on the Spectral Windows icon.
This allows you to define whether the
acquisition is to be over a single shot
window or an extended range.
The options from the drop down box are:
• SIMPLE = single shot
• MULTIWINDOW = extended range (1)
• CREST = extended range (2)
Page 14
Acquiring a spectrum - continued
Simple
This method allows a single shot spectrum to be acquired, with user defined integration
time and averaging.
1.
Click on the Spectral Windows
icon and choose SIMPLE from the
drop down menu.
2.
Ensure the central spectrograph
position
has
been
correctly
selected.
3.
Choose
required
4.
Choose
the
accumulations.
5.
Click on the Acquisition Options
icon to select a cosmic ray (random
spike) removal algorithm.
See
page 24 below.
6.
Now click on the Spectrum
Accumulation icon to start the
acquisition.
7.
The acquisition can be stopped at
any time by clicking on the STOP
icon (top right of screen).
the
integration
time
number
of
Multiwindow (extended range 1)
This method allows a spectrum to be acquired over an extended range, with a defined
integration time and averaging. The extended range is covered by taking a number of
individual single shot windows and ‘gluing’ these together. This procedure is fully
automated through the software.
1.
Click on the Spectral Windows
icon and choose MULTIWINDOW
from the drop down menu.
2.
WINDOW
In
the
SPECTRAL
PARAMETERS section, type in the
Start (“From”) and Stop (“To”)
positions.
3.
Choose what COMBINING MODE
options are required (see below).
Page 15
Acquiring a spectrum - continued
4.
Select the integration time and
number of accumulations from the
tool bar at the bottom of the screen.
5.
Click on the Acquisition Options
icon to select a cosmic ray (random
spike) removal algorithm.
See
page 24 below.
6.
Click
on
the
Spectrum
Accumulation icon to start the
acquisition.
7.
The acquisition can be stopped at
any time by clicking on the STOP
icon (top right of screen).
COMBINING MODE
The basic operation of MULTIWINDOW acquisition involves the acquisition of a number
of discrete spectral windows, covering the chosen extended range. In order to simplify
the process, it is possible to instruct the software to combine the individual spectra
automatically, resulting in just one spectrum over the chosen extended range.
• Combination after acquisition – checking this box
will result in the individual spectra being
automatically combined immediately after the
acquisition.
• Remove combinated objects – checking this box
will cause the individual spectral to be deleted
following the combination process, leaving just
the final combined spectrum.
• Adjust objects intensity – in order to remove any
possible artefacts (such as steps) being
introduced into the combined spectrum,
checking this box will cause the baselines of the
individual spectra to be adjusted prior to
combination, in order to give the best possible
result.
• Overlap – this defines the overlap between the
individual windows, in pixels. A typical value of
50 will give good results.
Note that individual windows can also be manually combined after acquisition simply
be clicking on the COMBINE button. The Remove combinated objects and Adjust
objects intensity check boxes will still function.
Page 16
Acquiring a spectrum - continued
CREST (extended range 1)
The Continuous Rapid Extended Scanning Technique involves moving the grating in a
number of very small steps, allowing the spectrum to be slowly built up step by step.
The advantage of this method is that any pixel to pixel variation in response can be
averaged out over the spectrum.
3000
Intensity (a.u.)
2500
Offset individual spectra
2000
1500
1000
500
Resultant spectrum
0
0
1000
2000
3000
Wavenumber (cm-1)
1.
Click on the Spectral Windows
icon and choose CREST from the
drop down menu.
2.
In
the
SPECTRAL
WINDOW
PARAMETERS section, type in the
Start (“From”) and Stop (“To”)
positions.
3.
Set the ACCUMULATION NUMBER. A
good starting value for this is 10,
but higher or lower values can be
used as desired.
For more
information see below.
4.
Set the SUB-PIXEL FACTOR to 1,
and SMOOTHING (%) to 0. Again, for
more information see below.
5.
Select the integration time and
number of accumulations from the
tool bar at the bottom of the screen.
Note that the accumulation number
set here is in addition to the
CREST ACCUMULATION NUMBER
already specified.
Page 17
Acquiring a spectrum - continued
6.
Click on the Acquisition Options
icon to select a cosmic ray (random
spike) removal algorithm.
See
page 24 below.
7.
Click
on
the
Spectrum
Accumulation icon to start the
acquisition.
8.
The acquisition can be stopped at
any time by clicking on the STOP
icon (top right of screen).
ACCUMULATION NUMBER, SUB-PIXEL FACTOR, and SMOOTHING (%)
Accumulation number – this value represents the number of individual acquisitions
contributing to any particular data point in the final spectrum. The higher this number
the better will be the signal to noise, the smaller the step size, and the more pixel to
pixel averaging there is.
Sub-pixel factor – setting this number to anything other than one will cause the
spectrometer to step by an amount less than a whole pixel value. The result is to
increase the number of data points defining a band (ie, 2 = twice the number of data
points, 3 = three times the number etc). Note that this will increase the total acquisition
time, and it is suggested that sub-pixel acquisition is only used over limited ranges.
6000
4000
sub-pixel = 4
3000
2000
5000
Intensity (a.u.)
Intensity (a.u.)
6000
sub-pixel = 1
5000
4000
3000
2000
1000
1000
0
0
510
515
520
525
Wavenumber (cm-1)
510
515
520
525
Wavenumber (cm-1)
Smoothing (%) – this allows control over the algorithms used for the CREST sub-pixel
acquisition, and relates to the response profile of an individual pixel to light. Typically
this value can be set to 0.
Smoothing (%) = 0
Smoothing (%) = 100
Page 18
Acquiring a spectrum - continued
Separate spectral regions and independent accumulation times
As well as defining a complete extended range for acquisition, a number of discrete
ranges can be acquired, for example, 200-1000 cm-1 and 2500-3500 cm-1. In this way,
only regions of interest need be acquired, and regions with no (useful) information can
be ignored, thus saving time. Additionally, the separate ranges can be acquired with
independent integration times, so that the resulting signal to noise can be optimised for
each window.
1.
Click on the Spectral Windows
icon and choose MULTIWINDOW or
CREST from the drop down menu.
2.
Check the boxes in the SPECTRAL
WINDOW PARAMETERS section to
select the number of individual
windows required, and type in the
Start (“From”) and Stop (“To”)
positions for each region.
3.
For CREST set the ACCUMULATION
NUMBER, SUB-PIXEL FACTOR and
SMOOTHING (%) values.
Note
that
for
MULTIWINDOW
acquisition the automatic combine
function does not operate when
acquiring discrete spectral regions.
However, the regions can be
combined
manually
after
acquisition, by clicking on the
Combine button.
4.
If independent accumulation times
are required for the individual
regions, check the Independent
Accumulation Times box, and set
the integration time for each region.
5.
Set the number of averages in the
toolbar at the bottom of the page.
6.
If using independent accumulation
times for each region, it is useful to
set the intensity scale to “counts
per second” (via the OPTIONS menu
in the toolbar).
OR
Page 19
Acquiring a spectrum - continued
7.
Click on the Acquisition Options
icon to select a cosmic ray (random
spike) removal algorithm.
See
page 24 below.
8.
Click
on
the
Spectral
Accumulation icon to start the
acquisition.
9.
The acquisition can be stopped at
any time by clicking on the STOP
icon (top right of screen).
Note that the automatic combine function does not operate when acquiring discrete
spectral regions by MULTIWINDOW, but the regions can be combined manually after
acquisition.
For other acquisition functions which can be used in conjunction with those discussed
here, please see the Other Acquisition Functions section on page 24.
Page 20
Acquiring Mapped Images and Profiles
Introduction
With suitable accessories on the LabRAM, it is possible to acquire XY mapped images,
and line (X or Y), depth (Z), time and temperature profiles.
The principal of these measurements involves acquiring an array (2D or 1D) of spectra,
with each spectrum acquired with a particular varied parameter (for example, position,
or time).
Variation
Description
Hardware requirement
X and Y position
map
motorised XY stage
X or Y position
line profile
motorised XY stage
Z position
depth or Z profile
motorised Z stage/piezo
X, Y and Z position
volume
motorised XY and Z stage/piezo
Time
time profile
no accessory required
Temperature
temperature profile
heating/cooling sample stage
In all cases, the spectral array (3D for volume, 2D for map, 1D for depth, line, time, and
temperature profiles) is saved as a single file, allowing fast and easy analysis of the
data.
The example image below shows the point by point nature of a Raman mapped image.
Page 21
Acquiring Mapped Images and Profiles - continued
Setting up a mapping/profiling experiment
With suitable accessories on the LabRAM, it is possible to acquire XY mapped images,
and line (X or Y), depth (Z), time and temperature profiles.
1.
On the microscope select the
objective to be used for the mapping
experiment, and select the same
objective in the software.
2.
If an XY map, XYZ volume, or line
profile is to be acquired, obtain a
white light image of the sample, and
define the area/line to be analysed.
Discrete points across a surface (right
mouse click to add and delete points)
Horizontal (X) line
Vertical (Y) or sloped (XY) line
Rectangular (XY) area
Polygonal (XY) area
Circular (XY) area
If a depth (Z), time or temperature
profile is to be acquired, simply
ensure that the sample is correctly
positioned beneath the microscope.
3.
Click on the Acquisition Data
Parameters icon in order to define
the mapping parameters.
Time
Temperature
X, Y and Z
(Pixel binning of CCD)
Page 22
Acquiring Mapped Images and Profiles - continued
4.
For each specific property, set up
the experiment by clicking on the
relevant icon. This will give you a
choice for how the parameters are
defined:
Don’t use – do not use that property
Defined size – defines the data
array size
Defined step – defines the step size
in s, oC, or µm.
The examples on the right show an
XY map set up, either by defined
size (indicating a final array of 20 x
20 points), or defined step (indicating
a step size of 10 µm in both X and Y
directions). If size is defined, step
will adjust accordingly, and vice
versa.
Defined size
Defined step
or
Area/Volume mapping
For area mapping X and Y dimensions
need to be defined. For volume, define
the Z dimension as well.
Line profile
For horizontal line profile, X dimension is
defined only.
For sloped line profile, X and Y
dimensions are active, but size and step
are defined only through the X dimension.
Time profile
For time profile, define time only.
Depth profile
For time profile, define time only.
Temperature profile
For temperature profile, define
temperature only.
Page 23
Acquiring Mapped Images and Profiles - continued
4.
(continued)
Further specific details:
For time profiles, the step size is in
seconds, and defines the time
between
the
start
of
each
acquisition. Note that the overall
acquisition time per data point must
be less than the step size.
For depth (Z) profiles, a negative
number in the variation box implies
moving the analysis point into the
sample (ie, -20 ! 20 µm will move
20 µm below the start position, and
step its way up to 20 µm above the
start position). The start position is
always defined as 0 µm.
For temperature profiles, a simple
ramp (ie, Defined size or Defined
step) doesn’t include a dwell
(equilibration)
time
at
each
temperature. In order to do this,
choose Arbitrary and click on Set
Up
to
define
the
required
temperature profile with dwell times.
Page 24
Other Acquisition Functions
Cosmic Ray (Random Spike) Removal
The CCD detectors such as those used on the LabRAMs are sensitive to other forms
of radiation, and in particular to random events known as ‘cosmic rays’. These can
interfere with acquisition of spectra by registering as very sharp and strong bands in
the spectrum.
Since the occurrence of a cosmic ray is random, it is extremely unlikely that a cosmic
ray will occur in exactly the same part of the spectrum in two or more consecutive
accumulations. Hence, it is possible to use a simple algorithm to detect random
events in a spectrum (as opposed to constant features, such as a Raman peak).
In LabSpec there are two algorithms to choose from.
Multi accum This is the more robust algorithm, and works by comparing two (or
more) accumulations within an acquisition. If a random cosmic ray spike is observed,
the spike will be removed. For this algorithm to work, the accumulation number
(averaging) must be set to greater than 2.
Single accum This algorithm attempts to locate a spike in a single accumulation
acquisition (ie, accumulation number = 1), by analysing bands for sharpness (width)
and intensity. This algorithm needs to be used with care, for very sharp Raman
bands can easily be modified. However, the algorithm works very well when looking
at broad features (such as Raman bands of amorphous materials, photoluminescence
etc).
Note that both algorithms can be used for SIMPLE, MULTIWINDOW, and CREST, single
point, mapping, profiling etc.
1.
Click on the Acquisition options
icon.
2.
Select which cosmic ray removal
algorithm to be used.
3.
Ensure that the accumulation
number
(averaging)
for
the
acquisition is set appropriately:
Multi accum ! 2 or more
Single accum ! 1
Page 25
Other Acquisition Functions - continued
Autosave
The Autosave function (via OPTIONS in the menu bar) automatically saves each
spectrum/mapped image/profile immediately the acquisition finishes.
Autosave on/off
Repeat an acquisition at
defined time intervals.
Directory path for file
save, with auto suffixing
folder name (DD=day,
MM=month, YY=year)
File save format (tsf =
standard labspec format)
File name for autosave,
with
auto
suffixing
(NN=number, HH=hour,
MM=minute).
Page 26
Other Acquisition Functions - continued
Autofocus
The Autofocus ensures an optimal focus is reached before an acquisition.
The Autofocus can work in three ways, depending upon the instrument configuration:
• Monitor total scattered light intensity with a pin diode – maximum = focus position
• Monitor laser spot size on video – smallest, brightest spot = focus position
• Monitor Raman intensity on CCD – maximum intensity = focus position
Click on the Acquisition Options icon
. Check the box to activate the autofocus,
and click on the Autofocus button to set up the autofocus function.
Depth
Image
and
Intensity Image – when
using
autofocus
for
Raman
mapping,
checking these boxes will
create additional images,
corresponding
to
the
focus depth position and
the
autofocus
diode
intensity in order to give
topographical information
about the sample.
If using the Z motor, set
the
start
and
stop
positions
for
the
autofocus
movement,
together with the step size
(all units in µm).
Negative number implies
moving the focus position
further down into the
sample
Use either the pin diode,
the
autofocus
video
display, or CCD readout
for calculating the focus
position.
Offset (in µm) to be
applied to the calculated
focus position.
Force the software to use
either the Z motor or the
piezo for autofocussing.
When using the CCD
signal,
specify
the
spectral range to be used
(in whichever units are
currently
set
via
OPTIONS in the menu
bar) and the integration
time.
Page 27
Other Acquisition Functions - continued
Autoexposure
The Autoexposure function allows the software to automatically adjust the integration
time to ensure a good signal to noise. An initial acquisition is made, using the
standard acquisition time set in the hardware tool bar. The quality of this spectrum is
then compared to preset autoexposure parameters, and if necessary the acquisition
time is increased or decreased to fit the required limits.
The Autoexposure is activated by checking the box in the main hardware tool bar.
Once activated, click on the Autoexposure icon
to adjust the parameters.
Saturation control - sets the upper limit for acquisition eg. 9,500 counts. If this
limit is reached it will calculate the acquisition time required to bring the
spectrum below this threshold. A minimum time value can be set so that the
system does not try to continually adjust this ad-infinitum.
Low level control - sets the values for low signals. The threshold value here
defines the minimum difference between the highest and lowest intensity data
points. With any measurement below this threshold value the software will
calculate the increased time required to achieve a suitable acquisition (to meet
the desired threshold value if entered) and set this as the resultant acquisition
time. Again, a maximum acquisition time is specified here, to prevent the
software attempting to make an infinitely long acquisition (for example, if there
is no signal.
Saturation threshold
30000
Intensity (a.u.)
25000
20000
Low level value
15000
10000
5000
400
600
800
1000
1200
Wavenumber (cm-1)
1400
1600
Page 28
Other Acquisition Functions - continued
Extended white light video imaging
With a motorised XY stage on the LabRAM, it is possible to montage a number of
individual white light images together, in order to obtain an image from a much large
area than would normally be possible.
1.
Set up the LabRAM for white light
illumination.
2.
Ensure the correct objective is
specified in the hardware tool bar.
3.
In the software, click on the
Extended video acquisition icon.
4.
Set the parameters for
extended image acquisition.
the
Number of images = total number of
images in X and Y dimensions to be
montaged.
Max size (pix) = the maximum pixel size of
the final extended image.
Overlap (pix) = individual images are
overlapped in order to ensure seamless
montaging.
Input the size of overlap
regions to be used.
Total size (µm) = display of the total size of
the final extended image.
5.
Click on the Start button to start the
acquisition.
Note that once an extended image has been acquired all the standard map/profile
experiments can be performed using this image.
Page 29
Section 3
The LabSpec Software for Data Analysis and Display
Page 30
Data Analysis Functions
The LabSpec software contains a wide range of data analysis functions, which are
briefly described here.
Arithmetic
Allows spectra to be have a constant value added/subtracted, or be
multiplied/divided by a constant value.
Additionally, two spectra can be added/subtracted/multiplied/divided.
Add/subtract and
multiply/divide by
constant value
Add/subtract and
multiply/divide
two spectra
Baseline Correction
Allows removal of a background (eg, fluorescence) from a spectrum.
Degree of polynomial
to
be
used
to
approximate baseline
Operation
allows
control
of
inserting/deleting
points
from
the
baseline
(Ins/Del),
adding or multiplying
by a constant value
(+const, *const) and
zooming in on a
region (zoom). Note
that +const and *const
only
work
when
Attachment is set to
No.
Type of baseline
approximation: either
polynomial
(data
points lie on a
polynomial curve), or
line segment (data
points joined by
straight lines)
Attachment: Yes = data points forced to
attach themselves to the spectrum, No =
data points can be freely placed anywhere
in window.
Once the baseline has been correctly approximated, click on Sub to subtract
the baseline from the spectrum.
Page 31
Data Analysis Functions - continued
Correction
Allows normalisation, zeroing, and correction of a spectrum.
Zeroes the spectrum (ie, subtracts a constant
value so that the lowest data point lies at
zero).
Allows a reference spectrum to be used for
correction purposes. Activate the spectrum to
be used, and click Get.
Normalise spectrum –
ie, adjusts the intensity
so
that the total
integrated area of the
spectrum = 100
In order to normalise to a particular band, activate a spectrum containing the
band, and click on Get. Adjust the two green cursors in the Correction
window to select the band to be used for normalisation.
Clicking on Norma will now normalise the active spectrum so that the
integrated area of the selected band = 100.
Page 32
Data Analysis Functions - continued
Profile
Allows a series of spectra to be bundled together into one file – a spectral
profile. Examples of profiles include line analysis, time and temperature
profiles, analysis of a series of slightly differing materials etc. Compiling
spectra into one profile can simplify subsequent analysis.
In addition, a profile in the X or Y direction can quickly be generated from a
Raman mapped image using this tool.
Creating a profile from individual spectra
1. Open the Profile window.
2. Activate the first spectrum to be added to the profile.
3. Click on Add in the Profile window.
4. Repeat the process with additional spectra to be added into the profile.
5. Once complete, close the Profile window.
6. Three windows will now be open:
Spectral_profile – overlapped data of all spectra
Profile – the profile displayed according to intensities between
the three cursors (R, G, and B).
Spectrum – spectrum at current cursor position
Creating a profile from a mapped image
1. Place the map cursor in the correct X (horizontal) or Y (vertical) position to
select the precise row/column from which to create the profile.
2. Click on Horizontal to create a profile across a row of the map (X
dimension), or Vertical to create a profile down a column of the map (Y
dimension).
3. A new window will be created showing the profile.
Page 33
Data Analysis Functions - continued
Smoothing and Filtering
Allows a spectrum to be smoothed in order to reduce noise, and other filtering
algorithms (such as spike removal) to be applied.
Once the parameters are correctly set (see below), click on the Filtr button to
apply the algorithm to the spectrum.
Defines which dimension is being
smoothed/ filtered. For a single
point spectrum this defaults to
Frequency but for maps and
profiles, other dimensions can be
selected.
Number of adjacent
pixels to be used
for
smoothing/
filtering.
Display of adjacent
pixels. The weighted
contribution from each
pixel can be set if
required.
Type of smoothing/filtering. For
spike removal choose Despike, for
a standard smoothing function,
choose average.
Fourier Transformation
Applies Fourier transform processing to the active spectrum. Using the IFT
functionm allows particular frequencies contributing to the spectrum can be
removed (for example, removing high frequency can reduce the random pixel
to pixel noise apparent in a spectrum).
Allow real time refresh of
spectrum with applied
DFT and IFT processing.
Process set up.
Apply DFT/IFT
processing
Zoom function – input
value to limit how much
of the FID is observed
(100% = full).
Page 34
Data Analysis Functions - continued
Peak Labelling and Band Fitting
Allows peaks within a spectrum to be quickly labelled, and for particular
bands to be fitted in order to accurately calculate band position, width,
amplitude and integrated area.
Peak Labelling
To activate the peak labelling algorithm, either click on the Search button, or
adjust the Height and Neighbour scroll bars. These two scroll bars control
parameters used within the searching algorithm for locating peaks in terms of
the peak size and proximity to other peaks. Adjusting these scroll bars will
allow the labelling to be optimised.
Real time refresh of
labelling on the spectrum
so that the effect of
adjusting
Height
and
Neighbour
can
be
monitored.
Check boxes allow the display to be controlled:
Peak label – display the peak label
Band sum – display the complete band sum used in the fitting
Band shape – display individual band shape for each labelled peak
Attachment – attach the peak label to the peak, or float at top of window.
Clear the labels
from the spectrum.
Label format – allows the
number of decimal places
to be set (1 = X.X, 2 =
X.XX etc)
Note: if a spectrum is saved with peak labels, the peak label information will
be saved with the spectrum for future reference.
Page 35
Data Analysis Functions - continued
Peak Labelling and Band Fitting (continued)
Band Fitting
1.
Click on Func in order to define the
algorithm to be used for the band
fit.
2.
Select the function to be used, and
add a baseline if necessary.
3.
Click on the peak Label icon, and
mark which peaks are to be fitted.
(left click = add, right click = delete).
4.
Once all the peaks have been
marked, click on Approx. and then
Fit.
5.
The results will be displayed on the
screen.
Note: if a spectrum is saved with the resulting band fit, the information will be
saved with the spectrum for future reference.
Fit parameters:
Iteration
number
=
number of iterations of the
fitting procedure to be
completed
before
stopping.
Deviation = if the χ2 value
changes by less than this
amount, the fit procedure
will
be
considered
complete.
Opens a new window
displaying the results of
the band fit for baselines
and peaks. (p=position,
w=full width half max.,
a=amplitude
or
max.
intensity,
g=Gaussian
contribution, s=integrated
area)
Allows
initial
start
parameters
to
be
specified and/or fixed.
Page 36
Data Analysis Functions - continued
Band Integration
Allows a fast integration of a band area.
1.
Select the green pair of cursors
2.
If the cursors aren’t visible on
screen, click on the Cursor
normalisation icon.
3.
With the cursors on screen, click on
the Integral icon.
4.
The area between the cursors will
be shaded, and the integration
results displayed.
The integrated area is divided into
two parts; the top section
(corresponding to the peak itself),
and
the
bottom
section
(approximating
background
contribution). The values displayed
correspond to the total integrated
area, and the area of the top and
botom sections:
Total = Top + Bottom
Page 37
Data Analysis Functions - continued
Object dimensions
Allows extraneous data to be removed from a spectrum/map/profile. For
example, if a spectrum includes data from 200 cm-1 to 3500 cm-1, but the
region of interest is only between 400 cm-1 and 800 cm-1, it is possible to
delete the data outside of this region.
1.
Activate the spectrum.
2.
Zoom in on the region that is to be
retained (‘extracted’).
3.
Click on the Extract button.
4.
Note that this change is permanent
if the file is saved, so it is
recommended that the extracted
data is saved with a new file name.
The spectrum/map/profile can also
be rescaled in all dimensions by
manually typing in the new limits
and clicking on Scale. The effect of
this can be reversed by clicking on
Expand.
Page 38
Data Analysis Functions - continued
Object generation
Controls visibility and behaviour of cursor components for map and profile
analysis by cursor and models.
If a particular component is deleted by accident, it can be returned by
unchecking and then rechecking the particular box in the Object generation
window.
Display R, G, B and G/B
components if checked
Display
spectrum
at
current cursor position
Cursors display intensity
without background
Display
model
result
components if checked.
Mapped images of models
Model spectra in window
Model fit result in window
ASCII multi-file save
Saves all open files (spectra/mapped images/profiles) in text format.
Browse for save destination
Remove the individual
spectra once saved.
Split a mapped
image / profile into
individual spectra if
checked (otherwise
data will be saved
in
a
multiple
column single file)
Includes
the
total number of
data points at
the top of each
row/column
Include X axis
data
(cm-1,
nm…)
Save as columns of
data if checked
(otherwise in rows)
Include point location information for mapped
images / profiles if checked. For example,
row/column will be headed by micron positions for
X and Y dimensions for a map
Page 39
Data Analysis Functions - continued
Custom Units
Allows units other than cm-1 and nm to be used. eV and kbar are preset, but
custom defined formulae can also be used.
Drop down box containing
custom units.
This
name
will
be
displayed in OPTIONS in
the menu bar.
x = wavelength in nm
Function
defining
custom unit
the
x = custom unit
Variables used in the
function formulae above.
Note that whatever unit is chosen from the Custom Units drop down box will
be displayed in OPTIONS (in the menu bar).
Page 40
Analysing Mapped Images and Profiles
Before any analysis is made it is recommended that the data is saved. In all cases,
ensure the window containing all the overlapped spectra (Spectral_Image or
Spectral_Profile) is highlighted, and save in the normal way.
A spectral image or profile is initially displayed with three active windows as below:
Spectral_Image (Spectral_Profile)
– containing all data of the
image/profile
Image (Profile) – the resulting
Raman image or profile
Spectrum – the spectrum at the
current cursor position
Note: the following descriptions are based upon analysis of a Raman mapped image,
but apply equally to analysis of a line, temperature, time or depth profile.
Analysing with Cursors
The initial method for analysing a mapped image or profile is to use three cursors (R,
G and B) to define regions. The image is generated by displaying the spectral
intensity between these cursors. For example, in the above diagram, the image can
be seen to comprise three coloured regions, which correspond to peak intensities in
regions defined by the cursors.
The three cursors are simply selected from the three coloured cursor icons on the
bottom right hand side of the Spectral_image and Spectrum windows.
If one or more of the cursors aren’t currently displayed on screen, they can be
returned by clicking on the Cursor normalisation icon
on the toolbar.
Using the cursors provides a fast method of getting useful information from the data.
However, in order to distinguish different components this method does require that
there are distinct, non-overlapping peaks which can analysed through the cursors.
Page 41
Analysing Mapped Images and Profiles - continued
Analysing with Modelling
With the modelling functionality, it is possible now to distinguish spectral components
that have a large number of overlapping bands, and look at the distribution of any
number of species (with the cursors, this is limited to just three – one for each cursor).
The modelling algorithms are based upon correlation fitting of known reference
spectra (the models) to the raw data. These model spectra can in fact be obtained
separately (ie, run spectra of known raw materials) or taken from the mapping data
itself.
1.
With the map data open, click on
the Model icon on the toolbar and
set if for Full search tree.
2.
If you wish to load in model spectra
which you have already acquired
separately, open up these spectra.
3.
Activate the spectrum you wish to
use as the first model component.
If you wish to take this model
component directly from the
mapping data, use the map cursor
to find the spectrum you wish to
use, and then activate the raw data
spectrum (light blue) in the
Spectrum window.
4.
Click on Get in the Model window.
5.
A new window will open up which
contains the model spectrum. The
purple cursors define which region
of the spectrum is to be used for
the correlation fitting.
6.
This process can now be repeated
as desired. Remember, if model
spectra are being taken from the
map itself, ensure the light blue
spectrum in the Spectrum window
is highlighted before clicking Get.
Page 42
Analysing Mapped Images and Profiles - continued
The Image window now contains both the information from the cursors and the
models.
If the data is in overlay mode (through FORMAT > VIEW in the toolbar), the image will
either display all the cursor intensities, or all the model intensities, but not both at the
same time. Which is displayed can simply be chosen by using the mouse to activate
any cursor indicator or any model indicator.
Cursor indicators (R, G, B and G/B)
Model indicators
The Spectrum window now shows not only the raw spectrum at the current cursor
position but also information about the correlation fit.
The coloured legends in the top right hand corner indicator the contribution of each
component to the correlation fit to provide a match with the raw spectrum.
The coloured spectra displayed comprise:
Light blue = raw data
Red, Green, Blue etc = models
Yellow = Fit
Raw data
Model 1
Model 2
Model 3 etc
Resulting fit
Results of fit
Page 43
Analysing Mapped Images and Profiles - continued
Displaying a map of band position, width, area etc
The data contained within a mapped image or profile can be analysed by a band
fitting routine, and the results can then be plotted to give a map showing band
position, width, area etc. Such analysis can be of use when investigating changes in
phase, stress/strain in materials and other such applications.
1.
With the map data open, in the
Spectral_image window use the
zoom tool to select just the area
which contains the bands which are
to be fitted.
2.
Click on the Data objects icon
and then the Extract button, in
order to remove all extraneous
data, and leave just the region of
interest.
It is best that this
extracted data is now saved with a
different file name, so that the
original map with complete data is
not lost.
3.
Open the Peaks & Bands window.
4.
Choose the band function you wish
to use for the fitting procedure, by
clicking on the Func. button. A
baseline can also be added at this
point.
5.
With a function selected, now select
the peak Label tool from the left
hand tool bar.
6.
On the Spectral_image window,
mark which peaks you wish to fit.
7.
Click on Approx. and then Fit to
start the fitting procedure.
Note: this could take a few minutes
to complete for a large map.
Page 44
Analysing Mapped Images and Profiles - continued
8.
Once the fit is complete, click on
Bands.
9.
A new window will open, containing
details of the band parameters
resulting from the fit.
These include information on the
baseline (if present) and all the
bands in the fit procedure. The
parameters are as follows:
p = peak position (cm-1)
a = amplitude (max. intensity)
w = full width half max (cm-1)
g = gaussian contribution (1=max)
s = integrated area of band
The band parameters shown are for
the spectrum at the current cursor
position.
10. Clicking on one of the parameter
labels will generate a new mapped
image, giving information about that
parameter.
For example, a map could be
generated showing the exact peak
position of a band, according to the
coloured scale (for example, dark
regions !
low wavenumber
position, bright regions ! high
wavenumber position).
Page 45
Data Display Functions
FORMAT in the menu bar allows a number of display settings to be adjusted.
Set the font used for axes etc.
Set the axes to be displayed, and
the colours of axes, gridlines and
spectra.
Adjust
the
spectral
view
(line,point,
bar),
overlay
behaviour, superimposition of
data for mapped images
Adjust 3D display of maps and
profiles.
Adjust the scale behaviour,
including fixed and auto scaling.
View
Display type for spectra (line,
point and bar) and images
(2D, contour and 3D)
Superimposition
of
map/profile data onto white
light image.
Colours/Axes
Check boxes allow axes,
titles, and gridlines to be
displayed. Colour palettes
allow their colours to be
changed if desired.
Axis titles (default values are
shown)
Overlay
behaviour
(single, multi, and multi
tiled)
Display all spectra,
selection, 100% or
25%)
Page 46
Display Functions - continued
3D Option – only accessible when the 3D View mode is chosen (FORMAT>VIEW).
Note that the only single components can be shown in 3D display.
3D display mode
Adjust the angle of the 3D
display.
Scale
Type the axis limits to
be displayed and press
return.
Rescale the X, Y and Z
axes to display all open
spectra.
Choose between the
scaling mode for the
axes.
Rescale all spectra
so that min and max
values are the same,
for easy comparison.
Auto scale
Used in conjunction with autoscaling, this
limits the autoscaling behaviour just for
increases in intensity, ie, scale will adjust if
intensity increases, but not if it decreases.,
Fix the scale to the
current
displayed
limits.
Page 47
Other Functions
Multi
The Multi function (via OPTIONS in the menu bar) allows whatever process is
performed on the active spectrum to be performed on all open spectrum.
For example, if a constant value is added to one spectrum, with Multi selected, all
the open spectra will have that constant value added to them. This can be a useful
way to save a number of files – with Multi selected, a save dialog window will appear
for each open spectrum, in the order they are displayed in the Objects list.
Axe + Text
If this option is checked (via EDIT in the menu bar), data saved as text format will
include both the X and Y axis data. With this option unchecked, only the Y (intensity)
axis data will be saved.
X and Y axis data saved as text format.
Y axis data only saved as text format.
Copy and Paste
The standard Copy and Paste functions (via EDIT in the menu bar) are slightly
expanded in LabSpec.
COPY TEXT = copy the spectrum in text format (one or two columns of data) to
paste into another application (Excel…)
COPY PICTURE = copy the spectrum window as an image to paste into another
application (Word, Powerpoint…)
COPY DATA = as COPY TEXT.
XY(Z) Stage
By checking this option (via OPTIONS in the menu bar) an additional window will
appear in the hardware toolbar, showing information about the current XY(Z) stage
position.
Reset all coordinates to 0.
Copy XY stage positions to
Scale Parameters window.
Disconnect / Reconnect
stage – allows manual
input of coordinate values.
Page 48
Other Functions - continued
Units
Units for the X axis can be specified as cm-1, nm and custom, and for the Y axis units
can be specified as counts or counts/s. Via OPTIONS in the menu bar.
Custom units are defined and chosen using the Custom units icon
toolbar – see page 39 for more details.
in the
More Objects
Up to 6 spectra can be chosen and activated through the coloured radio tags at the
edge of the spectrum window. However, More objects (via OBJECTS in the menu
bar) provides a full list of all open spectra, with coloured radio tag and file name.
Page 49
Other Functions - continued
Page Set Up
Page Set Up (via FILE in the menu bar) allows the print page to be configured,
including the spectrum itself, information from the acquisition, logo, and comments.
Properties
(including
zoom, colour / b&w,
frame, and orientation)
Add spectrum
Add text
Add signature
Add object
Save print page
Add file information
Note: to paste in an
image/logo,
use
EDIT/PASTE PICTURE in
the
main
LabSpec
menu bar.
Adjust font
Delete object
Print
Use current set up
as default
Close window
Page 50
Section 4
A Summary of Software Icons
Page 51
A Summary of Icons
Delete
Deletes active spectrum/mapped image/profile.
File Open
Open a file (spectrum, mapped image, profile, colour image, text, spc…).
File Save
Save a file (spectrum, mapped image, profile, colour image, text, spc…).
Cursor Normalisation
Bring the cursors back into window.
Rescale Window
Rescale the display that the entire spectrum (or all spectra) are displayed.
Print – see page 49
Print the current spectrum/mapped image/profile.
Information
Displays acquisition parameters relating to the active spectrum.
Spectral Windows – see page 13
Set the spectral range to be acquired (single shot, multiwindows, CREST).
Arithmetic – see page 30
Add/subtract/multiply/divide spectra.
Baseline Subtraction – see page 30
Model and subtract a background from a spectrum.
Page 52
A Summary of Icons - continued
Correction – see page 31
Zero or normalise spectra.
Profile – see page 32
Create a profile from a number of spectra, or from a mapped image.
Filtration – see page 33
Apply smoothing and filtering algorithms to the active spectrum.
Fourier Transform – see page 33
Apply DFT and IFT processing to the active spectrum.
Peaks and Bands – see page 34
Label peaks, and apply band fitting to spectra and mapped images.
Integral – see page 36
Calculate the integrated area between two cursors.
Palette
Adjust the colour/contrast/brightness settings for images.
Object Dimensions – see page 37
Extract a region of interest from the active spectrum.
Object Generation – see page 38
Control visibility and behaviour of cursors and models for mapping analysis.
Modelling – see page 41
Analyse maps and profiles using correlation fitting of model components.
Page 53
A Summary of Icons - continued
Video Readout – see page 11
Acquire a white light image of the sample.
Extended Video Acquisition – see page 28
Montage a number of white light images together for larger coverage.
Real Time Spectrum Adjustment – see page 13
Real time update of spectrum, with defined accumulation time – no averaging
Spectrum Accumulation – see page 13
Allows spectra to be acquired with multiple accumulations and averaging.
CCD Readout
Display the CCD image, with defined accumulation time.
Array Acquisition – see page 20
Acquire a mapped image, or line/time/depth/temperature profile.
Acquisition Data Parameters – see page 20
Set the parameters for an array acquisition (ie, step size, array size…).
Acquisition Options
Set up parameters for autofocussing, line scan and cosmic ray removal.
Detector Parameters
Parameters for CCD detector read out – includes temperature read out.
Scale Parameters – see pages 60 and 61
Set parameters for calibrating camera image and laser spot on screen.
Page 54
A Summary of Icons - continued
Spectral ID – see separate user manual
One touch link to Spectral ID database searching module.
ASCII Multi-file Save – see page 38
Save a batch of files in text format – for spectra, mapped images and profiles.
Custom Units – see page 39
Use custom units for X axis, including eV and kbar.
Stop Acquisition
Stop acquisition of spectrum/mapped image/profile.
Cursor
Use the cursors with the mouse.
Noise Filtration
Use the mouse to smooth a particular part of a spectrum.
Shape Correction
Amend a peak, or edit out a cosmic ray (random spike) with the mouse.
Zoom
Drag out the area to zoom with the mouse.
Horizontal Shift
Shift the X axis with the mouse.
Vertical Shift
Shift the Y axis with the mouse.
Page 55
A Summary of Icons - continued
Intensity
Rescale the intensity scale with the mouse.
Shift
Shift the X and Y axis scales with the mouse.
Add Constant – see page 30
Add/subtract a constant value to the active spectrum.
Multiply Constant – see page 30
Multiply/divide the active spectrum by a constant value.
Label – see page 34
Label a peak, or identify a band for band fitting.
Move Peak Maximum – see page 34
Adjust the position of a labelled/marked peak.
Fit Peak Width – see page 34
Adjust the band width of a labelled/marked peak.
Page 56
Section 5
Maintenance and Calibration Procedures
Page 57
Calibrating the LabRAM
Before acquiring a spectrum, the LabRAM needs to be calibrated. This is a simple
procedure involving two software parameters
ZERO
This is a number used to define the position of the zero order of the
spectrograph and can be thought of as the number of motor steps to
move away from a mechanical calibration sensor.
KOEFF
This can be thought of as the number of nm moved per motor step.
Part 1 - Calibrating the Zero order position
1.
Begin by selecting the grating you wish to calibrate and move the spectrograph
to zero order using the
icon in the Spectro. section of the hardware
toolbar.
Note that the two gratings are calibrated individually.
2.
Set the instrument up as follows:
•
•
•
•
•
Hole = 300 µm
Slit = 150 µm
Turn on the white light by reflection.
Ensure Camera beamsplitter is selected
Change UNITS to nm (via Options in toolbar)
3.
Now use the spectrum adjustment icon
to take a spectrum adjusting the
acquisition time or white light intensity until the signal level is around 10,000
counts.
4.
Press
.
Use the RED cursor to measure position of the band, remember that it is highly
unlikely to read exactly 0 nm. The band should be within +/- 1 pixel of Zero.
The way to measure the nm value of a pixel is to slowly move the RED cursor
and watch the position value increment. The value of one increment is the
value of 1 pixel, e.g. For a HR with a 1800 g/mm grating 1 pixel = 0.02nm.
5.
If the band is not at zero open the calibration window using the icon
. Now
adjust the ZERO parameter and watch the band position move, adjust until the
band is within +/- 1 pixel of 0 nm.
6.
Resend the spectrograph to zero order and retake the spectra. This will make
sure that your calibration changes have taken effect. Check the zero order
position again, and if necessary, make further adjustments.
NOTE:
The ZERO parameter should be changed in small increments e.g.
+5 or -5 at a time.
Page 58
Calibrating the LabRAM - continued
Part 2 - Calibrating the Raman spectrum
NOTE: This part of the calibration can also be done using a known
emission line from a Mercury or Neon lamp.
7.
Now change the UNITS to wavenumber (cm-1) and move the spectrograph to a
position at which you can monitor the Si Raman band (520.07cm-1).
8.
Insert your standard silicon sample and focus the sample in the normal way.
9.
Acquire a spectrum of the silicon sample (remember to remove the camera
beamsplitter and turn off the white light illumination) using the spectrum
adjustment icon
again - you should now be able to see the Silicon Raman
peak. Press
.
9.
Again use the RED cursor to measure the position of the band. It should within
+/- 1pixel of 520.07 cm-1. If it is not within this limit then adjust the KOEFF
value to move the position of the band.
Adjust the KOEFF by a small amount only (start by changing just the least
significant digit), and retake a spectrum each time you change the KOEFF to
monitor the effect.
NOTE: If the KOEFF is altered by a large amount it is advisable to
recheck the zero order.
10. Once you are satisfied with the calibration, close the calibration window and
ensure that you save the changes when prompted.
Zero and Koeff
values
Mfluo save
Click here to
save
the
current values
in the boxes
Mfluo restore
Click here to
reload
the
previous Zero
and
Koeff
values.
Page 59
Changing Laser Wavelength
1.
Open the top panel of the LabRAM, and remove the dust cover, in order to
access the top optics.
2.
Adjust/replace the items as displayed in the picture below:
Internal
laser in
External
laser in
If switching between two external
lasers, change over the clean up filter
If double notch/edge
filters are used,
change over the
second filter
Switching mirror –
up = internal laser,
down = all external
lasers
Change
notch /
filter.
the
edge
Change the notch/edge spacer
3.
Replace the dust cover, and close the top panel.
4.
Ensure all the push-pull bars are correctly chosen for the particular laser
wavelength to be used.
5.
Adjust the laser wavelength in the software:
Page 60
Calibrating the XY Stage Movement
1.
Insert the graticule, and focus on the markings .
2.
Ensure the same objective as is being used is
specified in the software.
3.
Capture an image of the graticule
4.
Using the “square/rectangle” mapping selection
cursor (selected from the bottom right of the
white light image), set the X dimension to match
exactly that of a known distance on the graticule.
5.
In the scale parameters box input the correct X
size. Having typed in the number, make sure
you press return, and then click “SET”.
6.
Repeat for the Y size (you may need to acquire
another white light image). Remember again to
press return having typed in the correct number,
and then click on “SET”.
7.
This procedure needs to be repeated for each
objective being used (ie, 10x, 50x, 100x).
If you are using a macro lens/objective (x4, x5) it
is likely that the graticule will be too fine to use.
A simple ruler with 1mm markings should be
sufficient in this case.
Page 61
Calibrating the Laser Spot Position
1.
Send the spectrograph to the reference diode
position by clicking on the right hand arrow.
2.
Open the confocal hole to 300 µm.
3.
Insert the standard silicon sample under the
microscope, and focus in the normal way.
4.
Turn off the white light illumination, but keep the
camera running.
5.
On the main control box, turn the reference
diode on using the left hand switch (marked
“diode”). [up = on down = off]
6.
You should now see the red diode spot on the
camera image. Adjust the microscope focus to
bring the spot to a tight focus.
7.
Close the confocal hole down to between 50 µm
and 100 µm (ensuring you can still see the diode
spot on the camera image). Adjust the focus
again.
8.
Stop the camera running.
9.
On the still image centre a single cursor point
-
Click on the discrete point cursor
-
Normalise
Page 62
Calibrating the Laser Spot Position - continued
10. Drag the cursor square to the centre of the diode
spot (it is best to zoom in on the spot using the
zoom icon
first)
11. Open the scale parameters box
12. Click on “SET” for the “Centr (xy)” row.
13. Click on “YES” to confirm the change to the
scale parameter.
Page 63
Installing LabSpec Software for Data Analysis
1.
Double click on the “LabSpec4XX.exe” to start
the installation procedure.
2.
Click on “next”
3.
The Installer will automatically choose a location
to save the LabSpec programme files. Click on
“continue”. If you wish you can amend the file
location, and then click on “continue”.
4.
The programme files are copied to the specified
location.
Page 64
Installing LabSpec Software for Data Analysis - continued
5.
The installer now searches for previously
installed LabSpec versions on the computer. If it
finds a previous version, it asks: “Would you like
to use the configuration files of the previous
LabSpec version?”
Click on “no”.
5.
The installation will be completed.
“Finish”.
Click on
6.
A Config window now opens.
“treatment”.
Click on
7.
At the prompt, click on “OK”.
Page 65
Installing LabSpec software for data treatment - continued
8.
Click on “Close”.
9.
When prompted, save the changes to the
configuration file by clicking on “Yes”.
10. A new folder labelled LabSpec 4.XX will appear
on the desktop, containing the following icons.
Using Spectral ID and Ascii Dump with LabSpec data treatment software
•
Exit the LabSpec software.
•
In the LABSPEC > DRIVERS directory on the hard drive, copy the files “toascii.dll”
and “search32.dll” and paste these into the LABSPEC > PLUGINS.
•
Click on “init” or “software reset” within the LabSpec folder on the desktop (and
click “OK” to confirm).
•
Restart the LabSpec software – two icons for Spectral ID (
Dump (
) will now be visible on the toolbar.
) and Ascii File
Page 66
Section 6
Contact Details
Page 67
Contact Details for Horiba Jobin Yvon
For further information in the UK please contact:
RAMAN SERVICE DEPARTMENT
For all enquiries concerning software, hardware, maintenance and service issues,
including lasers, heating/cooling stages, and other accessories.
RAMAN SALES OFFICE
For all enquiries about software and applications, upgrades, new accessories and
new instruments.
HORIBA Jobin Yvon Ltd.,
2, Dalston Gardens,
Stanmore,
Middlesex,
HA7 1BQ
Tel: 020 8204 8142
Fax: 020 8204 6142
http://www.jobinyvon.co.uk
Was this manual useful for you? yes no
Thank you for your participation!

* Your assessment is very important for improving the work of artificial intelligence, which forms the content of this project

Download PDF

advertising