HORIBA Jobin Yvon

Training LabRAM Spectrometer and LABSPEC Software

Gwnalle Le Bourdon HORIBA Jobin Yvon SAS

Ref: Doc.TM-LabLS/En

The LabRAM Description, functioning and optical adjustment P.2

1. Description of the Spectrometer different parts and optical path
2. Alignment : frequency calibration, verification of the laser alignement,
change of the exciting wavelength

- LABRAM TRAINING

3. Optimisation of the options available: choice of grating, choice of

objective, slit and confocal hole

LabSpec how to obtain and save spectra P.34

1. Acquisition modes
2. Multi-point analysis
3. Raman mapping
4. Kinetics measurements
5. Depth profiling
6. Configuration menu (acquiqition parameters storage)
7. Saving options : formats, autosave mode

LabSpec Modes of visualisation and treatment of spectres

1. Visualisation options (spectra, mappings, depth profiles)
2. Spectra treatments : baseline, smoothing, peak fitting, normalisation
3. Mapping treatment
4. Modelling

Use with samples

1
Ref: Doc.TM-LabLS/En

- LABRAM TRAINING

1st PART

The LabRAM

Description of the spectrometer

Adjustments and optical alignment

2
Ref: Doc.TM-LabLS/En

- LABRAM TRAINING

LabRAM Description:

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The different parts of the LabRAM :

Laser :
- internal : HeNe 17mW. Wave length: 632,817 nm

- LABRAM TRAINING

- external

Notch Filter :
1 Notch FIlter for each exciting wave length

Density Filters : To decrease the laser power

I = I0 x 10-D

By clicking her e, select one of the 6 neutral filters

with the optical densities 0.3, 0.6, 1, 2, 3 or 4.
filter [---] = no attenuation (P0 ), [D0.3] = P0 /2, [D0.6]
= P 0 /4, [D1] = P 0 /10, [D2] = P 0 /100, [D3] = P 0 /1000
and [D4] = P0 /10000
Microscope :
Illumination : 2 modes : transmission and reflection
Objectives : X10, X50, X100 (standard)

Confocal Hole: linked to spatial resolution

Close the confocal hole aperture

Initialize the confocal hole by closing

the hole and opening it to the previous
value.

Set the value of the confocal

hole aperture

Set the confocal hole to the maximum

value of aperture.
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Slit entrance : linked to spectral resolution

Spectrometer :

- LABRAM TRAINING

2 gratings
Movement controlled by Sin Bar

Detector : CCD

5
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Optical chamber :

- LABRAM TRAINING

Upper part
of LabRAM

Spectrograph
Lower part of
LabRAM

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Optical Path and stem options:

- LABRAM TRAINING

3
4

STEM

ACTION

ROLE

POSITION

Moves the beam

splitters BS12

Allows to choose
between a Raman
recording or a
visualization of the
laser sample

Pushed : visualization
with the camera
Pulled: Raman
measurement

Moves the block :

Allows to choose
Pushed : point mode
between

line

mode
Lenses L3, L4, L5, L6
Pulled: line mode
and

point

mode
and hole H2

Moves the mirror

M10

Moves the two

Choice between the
gratings 1800 g/mm two gratings
and 600 or 300
g/mm

Allows to choose
between the analysis
of a signal from the
microscope or from
the fiber optics
entrance

Pushed : point or
line mode
Pulled: fiber optics
analysis

Pushed : 1800 g/mm

grating
Pulled: 600 or 300
g/mm grating

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Ref: Doc.TM-LabLS/En

Laser Alignment

This should be carried out when there is a significant loss of signal (a test to verify the
confocality by using a silicon sample will reveal any misalignment see P. 29-30) or when
the spot laser shows problems with the centering or focus.

- LABRAM TRAINING

Aim : to ensure that the laser is perfectly centred on the confocal hole image.
In order to do this, use the internal diode which follows the return path of the Raman
beam.

With the Labram, you have the possibility to see the image of the confocal hole
projected on the sample when you switch on the laser diode for alignment.
The laser diode for alignment is placed inside the spectrograph, and, when the
grating is turned at an appropriate angle, the diode beam exits from the
entrance slit of the spectrograph and illuminates the confocal hole. If you put a
flat reflective sample under the microscope (like silicon or even a glass surface)
you can see the projection of the confocal hole on it.

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Ref: Doc.TM-LabLS/En

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Aligning the laser on the internal diode:

- LABRAM TRAINING

1. Turn on the internal diode:

2. Select the grating 1800 g/mm

3. Move the grating to the diode reference position

4. Position the camera for visualisation

5. Using the 100X objective focus on the silicon sample

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Ref: Doc.TM-LabLS/En

10

6. To locate the centre of the confocal hole easily, close to 100 m.

8. The diode spot determines the point which is used to align the spot laser.

There are two possible types of misalignment :

- LABRAM TRAINING

A centring problem:

A focusing problem:

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Should there be a significant disalignment of the laser, it is necessary to adjust the path by
using the mirrors as follows:
<Reference Laser 633nm>
You can adjust intensity by this mirror.
This mirror is very sensitive.

- LABRAM TRAINING

1,At first,you have to search maximum intensity by Z axis.

: Confirmation of intensity.
2,
3,Touch that mirror,and search maximum intensity value.
4,When you took a good value,you should check laser focus
after.

5,When you got bad focus,you can adjust by this mirror.

6,When you adjusted this mirror,you should check intensity
again.

<Others wavelength Laser>

You can adjust intensity by this mirror.
This mirror is very sensitive.
1,At first,you have to search maximum intensity by Z axis.
: Confirmation of intensity.
2,
3,Touch that mirror,and search maximum intensity value.
4,When you took a good value,you should check laser focus
after.

5,When you got bad focus,you can adjust by this mirror.

6,When you adjusted this mirror,you should check intensity
again.

A new confocality test has to be realised after the alignment (see P 29-30).
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Verifying the calibration frequency : zero order position of the gratings

The easiest way to test if your Labram is perfectly calibrated in frequency is to run a
silicon sample. You should find the Si 1 line at 520.7 cm-1.
If not, the zero order position of the grating has to be checked.

- LABRAM TRAINING

The gratings are driven with a sinus

arm that is linear in wavelength. In
fact the formula for the dispersion of
a grating is given by:

= [2cos(0)sin()]/nN , where N
is the number of grating grooves/mm;
n is the order and the angles 0 and

are represented in the picture. As

[2cos(0)]/nN is a constant, is
proportional to sin().

The coefficient ZERO is the mechanical position used as a reference for a value of
0nm. This corresponds to a number of steps of the motor between the switch of the
mechanical reference and the position 0 nm.
At the angle =0, the zero order of the grating must be centered on the detector. If it
is not the case, a constant shift in all the spectrum is observed (and thus on the silicon
band).
A small shift of the zero order can be produced by temperature changes. A shift of 6
pixels each 10 degrees of temperature change should be considered as normal.
Ref: Doc.TM-LabLS/En

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Adjusting the zero coefficient of the grating :
This is to be carried out for each grating.

- LABRAM TRAINING

1. Select the grating to be calibrated. Move the grating to the zero order position by
clicking on:

2. Select the following values for the confocal hole and slit entrance :
Hole = 400m
Slit = 150 m
Remove all samples and turn on the reflecting white light.
Let the white light enter the spectrometer by putting the camera beamspiltter.
Change the units of measurement to nm.
3. Use the icon :
to save a spectre and change the acquisition time of the intensity of
the light to obtain a signal around several thousands of counts.

4. Press STOP.
Use the red cursor to determine the position of the band. It should be
at 0 nm, at about +/- 1 pixel.

5. If the band is not at 0 nm, open the calibration window, by clicking on:
Before changing anything, note the values ZERO et KOEFF.

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Ref: Doc.TM-LabLS/En

- LABRAM TRAINING

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In the second menu Mfluo , change the value of ZERO :

Zero value [+] : Spectrum is going to minus direction.

Zero value [-] : Spectrum is going to plus direction.

Now adjust the ZERO and watch the band position move, adjust until the band is within
+/- 1 pixel of 0 nm

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Ref: Doc.TM-LabLS/En

- LABRAM TRAINING

15

6. Now change the UNITS to cm-1 and move the spectrograph to the position at which you
should monitor the Si Raman band (520.7cm-1) at the centre of the CCD (the central pixel
corresponds to the pixel for which one the frequency position is the same than the
spectrograph window value).

7. Insert your standard silicon sample and focus the laser in the normal way. Using the
spectrum adjustment icon
line. Press STOP.

again , you should now be able to see the Silicon Raman

8. Again use the RED cursor to measure the position of the band. It should within +/1pixel of 520.7cm-1.

9. Once you are satisfied with the calibration, close the calibration window and ensure that
you save the changes when prompted.

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Notch Filter
The Notch Filter is used to reject the laser and filter

- LABRAM TRAINING

the Rayleigh diffusion.

EASY

OPTICAL
DENSITY
7
6

POSSIBLE RAMAN
5
4

NO RAMAN

3
2

(degree)
0
15

10

T (%)
100

80

60

40

RAMAN
(cm-1)

20

0
0

50

100

150

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Characteristics of Notch Filters :

1 : filter references
2 : transmission of the filter
in function of the angle
3 : optical density in
function of the angle
4 : cut-off position in
function of the angle
5 : spectral edge width in
function of the angle
2

Edgewidth and cut-off definition

edgewidth
1
0
-1

-2

-500

4
4

5
5

80
Intensity (%)

- LABRAM TRAINING

Wavenumber
(
1)

60
40

50 %

20
0
-500

0
Wavenumber (cm-1)

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Aligning the Notch Filter.

Notch filter

- LABRAM TRAINING

Spacer

Spacer Number
Diameter (mm)
Angle of the
notch (degrees)

1
4
9,84

2
5
8,69

3
6
7,58

4
7
6,59

5
8
5,69

6
9
4,86

7
10
4,05

8
11
3,3

NB. Each Notch Filter must be used with the adapted spacer.
You can see the position of the cut-off of the notch filter looking at the white lamp in
transmission with a x10 objective. If the spectrograph is positioned on the exciting line
you can record a spectrum like :

The cut-off of the

notch
is
often
asymmetrical,
to
achieve a lower edge
of transmission. The
lowest wave number
that can be measured
reliably is around
100-120 cm-1.
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Hardware Section
Practical Advise Sheet n1 : Changing the excitation wave length

1. Removable mirror
2. Changing the Notch Filter (remember to choose a suitable spacer)

- LABRAM TRAINING

3. Software : enter the new wavelength value.

4. Choose a suitable grating (see p.20)

5. Position the spectrometer in the centre of the spectral window.

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Hardware Section
Practical Advice Sheet N2 : Choosing a suitable grating

The spectral resolution depends on :

- the grating
- the slit entrance

- LABRAM TRAINING

- the excitation wavelength

- the spectometers focal distance
- the number of pixels of the CCD

Parameters which can be optimised by the user:

- the grating
- the excitation wavelength
- the size of the slit entrance (in most cases a slit of 100 m is used)

Depending on the grating selected:

- the resolution differs
- the observed spectral range will differ

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- LABRAM TRAINING

Influence of the grating :

Gratings at fixed positions (1700 cm-1)

10000

1800 l/mm
600 l/mm

Intensity (a.u.)

8000

6000

4000

2000

500

1000

1500

2000

2500

Wavenumber (cm-1)

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Ref: Doc.TM-LabLS/En

707.9

22

12000

Different gratings same excitation wavelength

10000

953.7
939.4

966.1

923.5
908.8

4000

- LABRAM TRAINING

1800 l/mm
600 l/mm

929.9

6000
695.2

Intensity (a.u.)

8000

2000

700

750

800

850

900

950

1000

Wavenumber (cm-1)

To conclude:
Choice of grating depends on the application and aims of the measurement.

NB : other grating characteristics :

- The gratings are appropriate to a certain wavelength, meaning they have a reflection
maximum in certain spectral ranges.
The reflection of a grating is hence subject to the wavelength (1) and the light polarisation
(2)
TE light polarized parallel to the grooves
TM light polarized normal to the grooves

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IMPORTANT when working in the Near InfraRed!
- choice of grating (the grating 1800tr/mm is not suitable : optimised in visible and limited
mechanically)

950 tr/mm

- effect of the detectors response

Quantum efficiency, % .

60

633 - 787 nm

50
40
30

780 - 1030 nm

20
10
0
200

300

400

500

600

700

800

900

1000 1100

Wavelength, nm .

32000
30000

Pine excitation at 633 nm

Pine excitation at 780 nm

28000
26000
24000
22000

INT [a.u.]

- LABRAM TRAINING

Typical spectral response at 193 K .

20000
18000
16000
14000
12000
10000
8000
6000
4000
2000
0
250

500

750

1000 1250 1500 1750 2000 2250 2500 2750 3000

-1

Wellenzahl [cm ]

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Hardware Section
Practical Advice Sheet n3 : choosing the best objective
Numeric aperture of an objective : N.A. = n. sin(m)
m being the half open aperture
and n the refraction indice

- LABRAM TRAINING

Maximum diameter of the luminated spot is

NA = n sin ()

limited by diffraction phenomenas:

T = 1,22 x ( / NA )

eg : for a x100 objective of NA: 0,9

T = 1,22 x ( 632,81 / 0,9 )= 858 nm or 0,86 m

This resolution can be limited by the confocal

hole.

Lateral Resolution
Optical characteristics of the main objectives used:
Type of objective

10

50 X

50 X

100 X

100 X

LWD

MPlan

LWD

MPlan

33.4

48.6

53.1

64.2

Half aperture
max (m)
NA=n.sin (m)

0.25

0.55

0.75

0.8

0.9

W. D. (mm)

8.1

0.38

3.2

0.21

Spot diameter

3.1

1.4

1.03

0.96

0.86

632.8 nm

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Axial resolution field depth
The depth probed depends on the numeric aperture (NA) of the objective :
B High aperture : small volume studied
B Low aperture : large volume studied
The choice of objective will determine the intensity of the Raman spectra. Depending on
the sample type (opaque or transparent), the same objective will not have the same

- LABRAM TRAINING

behaviour.

1 Opaque sample

When there is almost no penetration of the laser in the sample, the Raman spectrum is
obtained mainly from the surface and its intensity is proportional to the collected flux. It
will be better to use a microscope objective with a high numerical aperture (x100,
NA=0.9) so that the solid angle (NA = nsin()) is bigger and you have a maximum
Raman signal.
The following drawing compares a x100 objective with a macro objective, which supports
this argument.

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Ref: Doc.TM-LabLS/En

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Silicon line intensity = f(NA)

100

obj 100x
90
80

Intensity (%)

- LABRAM TRAINING

70

obj 50x

60
50
40
30
20
10

obj 10x

0
0

0,2

0,4

0,6

0,8

Numerical Aperture

2- Transparent sample

If you have an homogenous sample, it will be better to use a microscope objective with a
big depth of focus (for example a x 10) so that it will collect the signal from a bigger
volume with a macro objective supports this argument.

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Hardware Section
Practical Advice Sheet N3 : Confocality

The principle of confocality

- LABRAM TRAINING

Confocal Hole

Rejected
Beams

Multilayered sample

Advantages :
(1) small increase in the lateral resolution
(2) large improvement in the axial resolution

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Ref: Doc.TM-LabLS/En

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Relationship between the aperture of the confocal hole (m) and the signal intensity (%)

- LABRAM TRAINING

This is also a test to verify the laser alignment.

Relationship between Depth (m) and

Intensity (a.u.) For 6 Confocal Hole
Apertures from 100 to 1100 m

Relationship between Confocal Hole

Aperture (m) and Axial Resolution (m)
(Full Width at Half Maximum)

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Ref: Doc.TM-LabLS/En

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Use of a quick confocality test to check laser alignment

Device
bar

- LABRAM TRAINING

Take a
spectrum

: Confirmation of intensity.

: Take in spectrum. (1 sec)

: Confirmation of intensity.

: Take in spectrum. (1 sec)

Device bar

Take a
spectrum

Check

Choose a 1000um spectrum.

: Select this icon.(Multiply Const)
Calculate the ratio of 1000um to 200um.

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Ref: Doc.TM-LabLS/En

- LABRAM TRAINING

30

Laser He-Ne (632.817 nm) :

-

The optimum confocality value = 60% with a confocal hole at 200 m/

confocal hole aperture at 100m

The laser is required to be adjusted if confocality becomes <40 %

Diode laser 785 nm :

-

Optimum confocality value = 35-40 % with a confocal hole at 200 m/

confocal hole aperture at 1000m

The laser is required to be adjusted if confocality becomes <20 %

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Harware Section
Practical Advice Sheet N4 : Taking measurements in Macro

- LABRAM TRAINING

With the Labram you have the possibility of working with macro-samples, gases and
liquids. For that you have to mount the accessory that is shown in the drawing below
which replaces one of the microscope objectives.

This macro-sampling device uses a 40 mm focal length lens, but other focal lengths are
available. You can even mount a microscope objective at the place of the 40 mm lens.
The advantage is that your laser beam exits horizontally and not vertically. An accessory
with a spherical mirror for double laser pass is also available for transparent material (see
below).

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Ref: Doc.TM-LabLS/En

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Troubleshooting : I cant acquire my spectrum

- LABRAM TRAINING

Check the following :

- Wavelength (nm/cm-1)
- Camera pull handle
- Z axis (Focus)
- Try reinitialization. (Hole/Slit/Wavelength)
- Is there some obstacle on light axis? (Paper)
- Checking of Power supply switches.

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Ref: Doc.TM-LabLS/En

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Troubleshooting : My spectrum look odd

- LABRAM TRAINING

Check :

That the internal diode hasnt been left on

That the white light is switched off

That the light in the room is switched off

Troubleshooting : My spectrum have a weak intensity

Check :

That there is no density filter on the laser beam

That the objective is clean

If necessary, clean the obhective with a mix 5% ether / alcohol. Soak the lens and place it
in an ultra sound bath.

Aligning the laser on a silicon sample (check the spot is centred and there is no
major focusing problems)

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Ref: Doc.TM-LabLS/En

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- LABRAM TRAINING

Second Part

LabSpec

Acquiring and saving a spectrum

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Ref: Doc.TM-LabLS/En

35

5 Practical Advice Sheets for acquiring :

- a spectrum in point mode
- spectra at multiple points in the sample
- a Raman mapping
- kinetics

- LABRAM TRAINING

- an in-depth profile

Precautions before recording spectrum :

Be sure that the laser power will not harm the sample ! Use density filters to decrease the
laser power in case the sample is sensitive to laser heating.

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36

Practical Advice Sheet N1 : recording a spectrum / point mode

1. Choosing the grating
Select the grating from the LabSpec software (LabRAM : a message will appear asking to

- LABRAM TRAINING

choose the correct stem position for the grating)

Click here to select one of the available spectrograph gratings
Do not forget to initialize the spectrograph after a grating change

Choose the grating with the use of stem 4.

In case the grating has not been used before, check its zero order.
Move the spectrograph to the
zero order position (0 nm)

2. Focusing the laser

Using either the video monitor or the video image function within LabSpec

3. Positioning the spectrograph

Centre the grating at the desired position

Enter here the spectrograph

position value

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Ref: Doc.TM-LabLS/En

37

4. Define the size of the slit (100m) and that of the confocal hole

5. Define the parameters exposure time and number of accumulations

Number of accumulations

- LABRAM TRAINING

Exposure time in seconds

3 possible acquisition modes

Type of acquisition
Simple window
Adjustment

Multi-window
Accumulation Mode

Parameters to define

Note

- Acquisition time

The previous

- Central position of the spectral

spectrum is

zone

automatically erased

- Acquisition mode
- Number of accumulations
- Complete spectral mode
(Multi-window)

Scanning Mode
Continuous (Kiefer
Scanning mode)

- Acquisition time
- Kiefer Scan parameters
(Spectral zone and number of
accumulations)

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38
Principle of the different acquisition modes :
Generally to get the whole Raman spectrum, multiple spectral windows are required. This
is achieved by moving the grating position in the spectrometer in some manner so as to
move the specific part of the spectral range which illuminates the CCD.
The first option is to use discreet spectral windows, which are glued automatically. (see
multi window acquisition option).

60000

50000

Intensity (a.u.)

- LABRAM TRAINING

Multi-Window Mode

40000

30000

20000

10000

0
500

1000

1500

2000

2500

3000

3500

Wavenumber (cm-1)

Kiefer Scan Mode

The method used by the new Kiefer scanning works in a different way and consists of
shifting the spectrum step by step so that each individual spectral element is detected
several times by the detector, rather than as a discreet spectral window.
The software and hardware can scan the spectrometer through a defined spectral region,
off-setting each subsequent acquisition viewed by the CCD detector to a certain amount.
This can be by a large number of pixel steps or even a sub-pixel value, depending upon the
required effect.
(i) Mode (I) - Larger Pixel overlap values :
In mode (I), a larger number of pixels is chosen as the offset, and an averaging effect is
produced. For this method the operational principle is that a datapoint ( in cm-1 ) is seen by
a number of different pixels on the CCD detector, and the average signal for this datapoint
is calculated.
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39

3000

Intensity (a.u.)

2500

2000

1500

500

500

1000

1500

2000

Wavenumber (cm-1)

(i) Mode (II) - Sub pixel offset.

The second way of using the Kiefer CREST scan is to use very small overlaps in the
spectrum, which provides an enhanced band definition.
Here a shift in position, entered in the sub Pixel value box, will move the grating
position by an amount less than one pixel on the detector. - A Sub Pixel.
By selecting a sub-pixel value (a value of 1 is for full integer values and hence
deactivates this mode), the step selected will increase the number of data points
obtained for the spectrum. Hence, a 2 subpixel value will give twice the number of
data points, 3 subpixel value three times and so on.
If you consider a usual acquisition, you have 1024 pixels (and 1024 data points).
Using the subpixel arrangement of a factor of 2, will give 2048 data points.
Whilst this operation does not affect the actual spectral resolution which remains
defined by the spectrometer focal length, grating and entrance slit, it does provide a
better band definition for Raman bands and can hence provide a better basis for
analysis of band shape and position for a given instrument.

8000

Intensity (a.u.)

- LABRAM TRAINING

1000

6000

4000

2000

39

0
428

430

432

434

436

438

Wavenumber (cm-1)

Ref: Doc.TM-LabLS/En

40
How to use the Kiefer Scanning Modes?

- LABRAM TRAINING

indicates the Kiefer scanning modes, by clicking on this, the window for the
Kiefer Scan will open:

start point is the starting point for the spectrum you wish to have (in cm-1 or in
nm)
finish point is the end point for the spectrum you wish to have (in cm-1 or nm)
accumulation number activates the first mode of the Kiefer scan used to
generate extended spectral regions and for spectral averaging, Mode (I)
The value represents how often a spectral data element will be detected (any value
can be entered) ie. the larger the number the greater the averaging.
sub-pixel factor defines the linearized data point (maximum is 6).
This is used to generate the higher definition mode. Again the greater the value, the
greater the definition, Mode(ii).

It can be considered that there are three cases for using the Kiefer scanning:

reduction of spectroscopic phenomenon ( on a wide spectral domain):

1) just enter the start point and finish point values,

2) select the accumulation number required
3) and set sub-pixel factor to 1.
4) Press start and the software drives the spectrograph, acquires the data and
linearizes the spectrum.
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The selection of the accumulation number depends on the required quality of the
end spectrum. The higher the number, the better the quality.
Integration time : for instance, if integration time T for classic acquisition is selected
and N accumulation for the Kiefer Scanning is also selected, the integration time
should be changed to T/N before starting the Kiefer Scanning.

- LABRAM TRAINING

- Improving the definition of the line shape ( on a short spectral domain) :

1) just enter the start point and finish point values,
2) set the accumulation number to 1
3) and select the sub-pixel factor required
4) Press start.
For the same reason as above, change the integration time to a lower value.

- A combination of both modes is possible but this is not really recommended as

this method will take time, especially if you require a complete Raman spectrum
with a definition four times better!

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42

Practical Advice Sheet N2 : recording spectra in several points in the

sample (Multi-points analysis)

- LABRAM TRAINING

LabRAM equipped with a motorised XY table

1.

Focusing the laser on the sample

2.

Recording the video image on the sample surface

Stop the acquisition of the continuous video

3. Define the measurement points with the cursor

4. Define the acquisition parameters :

- grating
- acquisition time and number of accumulations
- spectral zone multi-window
- confocal hole aperture and slit entrance

5. To start an acquisition use the icon spectral image

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43

Practical Advice Sheet N3 : recording Raman Maps

LabRAM equipped with a motorised XY stage

Initially, determine the best measurement conditions (acquisition time and number of
accumulations, confocal hole and slit apertures, density filter, spectral range) by

- LABRAM TRAINING

acquiring some few spectra in different areas of the sample. This allows one to optimize
the parameters and to prevent from detector saturation.

1. Displaying the sample with white light

Verify that the objective selected in LabSpec corresponds to the objective used.
Recording the video image

2. Open the window Acquisition Options

Select table in the sub-menu X-scanning and Cursor in scanning area.

Define the value in Refresh Time :which determines the period of time between the
two updates displaying the spectrum recorded on screen. In case of a long mapping
increase this time (eg. 3600 s) in order not to lose time with the use of display updates.
Close the Acquisition Options window.
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44
3. Choosing the cursor type to define the zone analysed
- a-

sloping line

- b-

horizontal line

- c-

rectangle

- d-

elipse

- e-

polygon

- LABRAM TRAINING

3. Selecting the parameters of the measuring zone

Open the Acquisition Data Parameters window
- Click on X and Y to define either the number of measurement points or the steps
between two points (m), then the software calculates automatically the second
parameters.

4. Define the acquisition parameters:

- grating
- acquisition time and number of acquisitions
- multi-window spectral zone
- Apertures of confocal hole and slit entrance

5. To start an acquisition, use the icon spectral image

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45

Practical Advice Sheet N4 : recording Kinetics

This option enables you to take a series of spectra or spectral images as a function of time.
Follow the same procedure as the one to record point by point spectra or spectral images.

- LABRAM TRAINING

But you also have to choose:

In the Data size window (select the time parameter, there are two types of boxes, the
first is the number of measurements and the second is the time interval between them).
Please refer to the Index section
Be careful, because the acquisition time is included in the interval of time between two
measurements.
To start an acquisition, use the icon spectral image

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46

Practical Advice Sheet N5 : Recording a profile in depth

Focus the laser on the sample,

Take a video image and freeze it with the icon,

- LABRAM TRAINING

Choose the cursor,

Select the time of exposure,
Select the
In the window Data size , select the Z scanning parameters.
- Please refer to the Index section
Press the icon
to start the acquisition

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Ref: Doc.TM-LabLS/En

47

Saving the measuring options

- LABRAM TRAINING

Configuration Window

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48

Saving spectrum

Save one of the displayed spectra, by activating the spectrum window, select the
desired spectrum by pressing the radio button, then choose one of the following
methods:

- LABRAM TRAINING

Click on the following icon:

then enter the file name, its location and format.
From the file Main Menu, select the Save As option, then enter the file name, its
location and format.
Before saving, it would be useful to complete the information list about the spectrum
(operator, laser power, etc...) by pressing the icon
NB. Some parameters (hole, slit, spectro, grating, time, accum, date) are automatically
updated.
LabSpec file formats.
The following is a list of the various file formats that LabSpec supports to save the
acquired spectra or images:
Dilor ASCII format (*. ms0): This format is used for single spectrum.
Extended Tiff
- (*.tsf):
It is a specific format for single spectrum and
LabSpec software.
- (*.tvf):
It is a specific format for spectral image.
Standard Tiff (*.tiff): Standard TIFF format for image.
Text format (*.txt): This is an ASCII mode format which uses two columns:
wavelength/Wavenumber and intensity, without header.
Spectra Calc format (*.spc):
processing softwares.

This format is used by SpectraCalc and Grams

Important comments:
- It is possible to save several spectra one after the other by using MULTI, (do not forget
to de-select after use) icon:
- when saving as txt, remember to activate Axe+Text
- Saving images:
The save procedure always saves the content of the active window. To save an image,
activate the window that contains all the spectra. If the Spectrum: Point window is
activated, only this spectrum will be saved, and not the whole spectral image.
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49
Saving in ASCII Format

- LABRAM TRAINING

This module permits to save all the spectra (or the spectra of a Raman image) of a window
as ASCII files (the maximum spectra saved in one shot is 64)

Destination path : indicate the address of the directory where the spectra will be saved.
Conversion option :
Different conversion options are available:
to remove the converted objects of the LabSpec window.,
to split the activated image to spectra that will be saved in ASCII
format,
to saved the ASCII file in two columns or two lines
to write the frequency values and/or the position values.
To activate the saving of the spectra press the button : SAVE ALL

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50

- LABRAM TRAINING

Auto Save

- You can choose the directory where the spectra will be recorded:

Yo u can choose a name

You can choose to use

the day, the month and the year as the name
MAXIMUM 8 CHARACTERS
- You can choose the file name of the recorded spectra:

Yo u can choose a name

You can choose to increment

the name by number, hour and minutes
of the acquisition.
MAXIMUM 8 CHARACTERS.
- You can choose the format of the files:
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51
Two options are available:
You can choose to work with the Auto Save and Auto Repeat options:
For this the delay between two acquisitions (eg: 10 seconds) must be chosen. Then after
clicking on the
icon the software takes an acquisition and repeats every 10
seconds, saving the spectra automatically.

- LABRAM TRAINING

However it is possible to work only with the Auto Save option:

- When the icon
it.

is selected, the software takes an acquisition, stops and records

- When the icon

is selected (which is used for spectral imaging, time scanning...)
the software takes a spectral image and at the end of the acquisition will be recorded.
MAKE SURE TO CHOOSE THE TSF FORMAT FOR THE SPECTRAL
IMAGES.

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- LABRAM TRAINING

52

Third Part

LabSpec

Processing Spectra and Raman Mappings

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53

Baseline Correction

Automatic
calculation

- LABRAM TRAINING

Additional/Subtraction
Clears the points
of the baseline
Saves as a
spectrum
To save the
baseline
Normalise the
spectrum
(maps)

The main procedure is to build up a baseline point by point, to best fit the spectrum and
then subtract it from the spectrum.
If you have to subtract the baseline from a spectrum, activate this spectrum.
If you have to subtract the baseline an image, activate the spectral image window.
Verify that the option Ins/Del is active (in the operations box)
Select the type for interpolation: it can be linear or polynomial (in the box type). For
the polynomial interpolation, you can also choose the degree of the polynom (in the
degree box).
You can select with the mouse in the spectrum window (validate: left button, delete:
right button) the points for the baseline computation
Hint:
(for the images, when you press the icon
average spectrum of the image is opened automatically).

, a window that contains an

Activate the option attachment , to fit at best the baseline to the spectrum.
Hint:
An automatic procedure can also be used for the computation of baseline:
button Auto
Press the button SUB .
NB: The baseline can be saved as a spectrum: by selecting the CONV button
Ref: Doc.TM-LabLS/En

53

54

Correction

- LABRAM TRAINING

Main use for processing images:

Enter the weakest value for the spectra as 0 and normalisation.

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55

Profile

2 main usess :

- LABRAM TRAINING

- (1) to create a profile file with several spectra separately saved and required to be
treated as images
- (2) to extract the profiles from a map.
(1) Create an image with several spectra : use the Add function after having activated the
spectrum into which the profile is to be added.

(2) Extract a profile from a map, either by following the horizontal or vertical lines.

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56

- LABRAM TRAINING

Peak-fitting

Preparation : Remove spikes on the spectrum

NB: in the case of an image, this modification must be carried out for each spectrum. If a
spectrum has a spike, remove this spike in the window Spectrum : Point and Press
button R to insert the corrected spectrum in the Raman mapping.

Step 1 : If only one part of the spectrum is of interest, first extract this zone by using the
window:

(if it is a spectrum or image)

Then, define the approximative positions of the bands

- by an Auto search (height parameter and neighbour to adjust)
- by using the following icon

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57

- LABRAM TRAINING

Step 2 : define the function for fitting/add a baseline

It is possible to choose a function independently for each band.

Step 3 : Define the parameters: number of iterations and deviations

Then start the peak fitting operation by clicking on
Then on

Step 4 : Visualise the reults by activating Band sum and Band shape.

Then for a table click on

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- LABRAM TRAINING

58

p : frequency position of the band.

a : amplitude of the band.
w : width at half maximum.
s : surface of the band.
g : Gaussian/Lorentzian ratio.
NB : it is possible to save these results by using the SAVE function
NB. : it is possible to fix the value of a parameter

Step 5 : with images

It is possible to draw up a map for each of the predetermined parameters by pressing keys
p , a , w , g or s .

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59
Examples of Raman mapping based on band Intensity/Frequency/Width (Thanks to Mr
Mermoux) :

- LABRAM TRAINING

Length Y (m)

200
300
x1000

400
50

500

intensit

600
400

500

600

Length X (m)

15000

1333.0

1333.0

1332.8

1332.8

1332.6

1332.6

frquence
10000

2.4

5000

2.2

Images : 4500 pixels

(pas de 5 m)
2h30 acquisition time

0
1200

largeur

1400

Backg

intensity
x1000

200
150

40

100

10 m

20

50

TEM

1340
10
1335
5
1330

frequency

width

100 microscope objective

Cathodoluminescence

Point spacing: 0.7 m

exc.=514.5 nm

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60

Modelling
It is the decomposition of each original spectrum of an image as a sum of model
spectra.

Choose the spectra that will become MODELS , which can be already saved on the

- LABRAM TRAINING

disk, or they can be particular spectra that have been extracted and saved from a spectral
image.
If the spectrum is a particular and extracted from the image and save.
Do the same with any other interesting spectra.
Load all the selected spectra and put them overlapped in a window (for that use the
option Behavior in the menu view in the FORMAT menu).
Activate the first spectrum selected and press the button get. Do the same for all the
selected spectra: a window is then created with all the model spectra .
In the single spectrum window, there are:

- The models with relative intensity.

- The experimental spectrum in particular point of the image.
- The result of the fitting.

You will see in the mapping window additional pictures that correspond to each one of the
model .
You can overlap them to see the relative contribution of each of the spectra in a particular
point of the image (for that use the option Behavior in the menu view in the
FORMAT menu
You can save a whole spectral image with the model compounds choosing the tvf
format. When loading this image, the decomposition will automatically appear.

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Ref: Doc.TM-LabLS/En