Raman - LabSPEC 5

CONFIDENTIAL
2009 HORIBA, Ltd. All rights reserved.

LabSPEC 5

CONFIDENTIAL
Notes
This tutorial is based on ARAMIS
LabSPEC operates ALL HJY Raman microscopes including
LabRAM HR, ARAMIS and XploRA.
LabSPEC comes with two icons: LabSPEC and Config.
LabSPEC icon opens the main software to operate the system
Config icon opens the configuration window to configure the system
LabSPEC is installed in C:\Program
Files\HORIBAJY\NGSLabSpec
Manuals are stored in C:\HORIBA Jobin Yvon\Manuals Aramis
Annotations
Names of icons, buttons and menu items are denoted in bold.
Names of windows are denoted in italic.
Names of columns, rows or entries in a window are denoted in all
CAPITAL.

CONFIDENTIAL
Overview Main Window
File I/O Data Information Experiment setup Data processing

Data acquisition
Axes and Cursors Stop
Laser selection
Neutral density filter Exposure time

Number of accumulation
Confocal hole diameter
XYZ positions
Slit width
Spectrometer Laser block/unblock
Grating System setup

Objective lens
4 CCD temperature monitor

CONFIDENTIAL
Overview Icons and Tool Bars
Right click on the background

Popup menu opens
Toggle between Big icons and Small icons
Turn on/off Left, Right and Bottom tool bar
Big icons
Small icons

CONFIDENTIAL
LabSPEC Window Types
Each window comes with
A heading Window Name: Data Name
Overview
Unique set of utilities on the left tool bar

Unique set of popup menu activated by right clicking the mouse
Video: Displays video or extended video images. Captured images
are saved to *.ngv format
Spectrum: Displays individual spectra. Saves to *.ngs format
SpIm: The parent window for mapping data. Saving from this
window saves all data and all information data acquisition
parameters, data processing histories, data processing results and
child windows.
Point: A child window of SpIm that displays a spectrum at the selected
point
Map: A child window of SpIm that displays an intensity map of the
selected spectral region
6

CONFIDENTIAL
Overview Video Window
Utilities for
Data name
Video Window
Window name (Video)
Popup menu

CONFIDENTIAL
Overview System Setup
System setup
One the best features of HJY Raman microscope is the
versatility. Setup button opens Aramis window that
allows switching between system modes.

CONFIDENTIAL
System Setup
Measurement Polarization
Location Modulates polarization
MICRO: measurement of laser and Raman
Overview
thorough the signal for polarized

microscope. Raman spectroscopy
RETRO: macro
measurement,
Measure
backscattering
Switches between point
MACRO 90: macro
mapping and line
backscattering, right
scanning mode
angle scattering
Visualization UV/Visible
VIDEO ON/OFF: live Switches between UV
video on/off and Visible laser beam
MICRO: internal paths
camera visualizing both
sample and laser spot
TOP: external camera
Detector
visualizing sample and
Switches between CCD
in sync with ocular
(1st detector) and
observation
PMT/IGA (2nd detector)
FTIR
9

CONFIDENTIAL
Axes, Cursors and Data Information Axes and Cursors
Rescale both X- and Y-axis Center cursors

Brings cursors to the center of window
Rescale Y-axis
10

CONFIDENTIAL
Axes, Cursors and Data Information Data Range
Spectrum
SpIm
Data range window shows range and size of each dimension of data.
Depending on the type of window, which is determined by the dimensions of data, when data set changes,
Data range window changes accordingly.
Data set can be truncated by altering FROM and TO, and then selecting Extract button
11

CONFIDENTIAL
Axes, Cursors and Data Information Parameters
Parameters window shows all

pertinent information about the
data set
Acq tab displays system Custom tab displays History tab displays data
parameters in data collection customized comments for processing performed on the
sample and data collection data
12

CONFIDENTIAL
Video Real Time Display and Acquisition
Video -30
Opens Video window and display real-time

video image of sample AND/OR laser spot. -20
The last image is captured and can be saved

-10
(*.ngv)
Y (m)
0
10
20
4 m
30
-40 -20 0 20 40
13 X (m)

CONFIDENTIAL
Video Extended Video Acquisition
Extended Video
Requires a motorized xy-stage.
Selecting the icon opens the Extended Video window for setting up the
extended video acquisition.
Start Video to open Video window and start recording extended video.
14

CONFIDENTIAL
Spectrum Laser Selection
Laser selection
As a new laser is selected, matching rejection filter must be also
selected, and spectrometer position is re-calculated to reflect the
change in Raman range.
15

CONFIDENTIAL
Laser Selection
Conventionally
Blue lasers: 400 500 nm
Spectrum
Green lasers: 500 600 nm

Red lasers: 600 700 nm
NIR lasers: > 700 nm
Choose the laser that minimize the fluorescence
background. Ordinarily, Raman spectrum is laser
wavelength independent while fluorescence background
is.
Choose for spectral resolution, spatial resolution or
speed of measurement.
Under otherwise identical conditions, short wavelength
lasers yield stronger Raman scattering, higher spatial
resolution and lower spectral resolution.
16

CONFIDENTIAL
Laser Selection and Fluorescence
70000 25000
60000
20000
Spectrum
50000
Intensity (a.u.)
Intensity (a.u.)
15000
785 nm
40000
10000
30000 532 nm
5000
20000
0
10000
500 1000 1500 2000 2500 3000 500 1000 1500 2000 2500 3000
Wavenumber (cm-1) Wavenumber (cm-1)
35000
Laser selection to avoid the
30000
fluorescence
25000
Intensity (a.u.)
633 nm
20000
15000
10000
5000
500 1000 1500 2000 2500 3000

Wavenumber (cm-1)
17

CONFIDENTIAL
Spectrum Grating Selection
Grating
As a grating is selected, spectrometer position is re-calculated to
reflect the change in dispersion
18

CONFIDENTIAL
Grating Selection
A higher groove density grating provides higher spectral

resolution and more data points per cm-1 for more
Spectrum
accurate results.
A higher groove density grating disperses light over a
wider angle, lowering the per pixel per second intensity
and shortening the spectral range at one position
requiring longer measurement.
It is recommended to start with a low groove density
grating and move up as needed
19

CONFIDENTIAL
Grating Selection and Spectral Resolution
600
500 Lidocane, 1800 gr/mm

Spectrum
Intensity (cnt/sec)
400
300
200
100
500 1 000 1 500 2 000 2 500 3 000 3 500

Raman Shift (cm-1)
3 000
Lidocane, 300 gr/mm

2 500
Intensity (cnt/sec)
2 000
1 500
1 000
500
500 1 000 1 500 2 000 2 500 3 000 3 500

Raman Shift (cm-1)
20

CONFIDENTIAL
Spectrum Confocal Hole
Confocal hole
Reducing the confocal hole diameter rejects out-of-focus signal
including fluorescence, stray light, signals from a container and
signals from out-of-focus matrix. This enables the acquisition of a
depth profile
21

CONFIDENTIAL
Confocal Hole
The main role of the confocal hole is to reject out-of-

focus signal for high spatial (especially axial) resolution.
Spectrum
The main role of the slit is to reduce the size of image at

the monochromator for high spectral resolution.
For micro measurement, start with 400 m confocal hole
and decrease as needed. For macro measurement,
open the confocal hole up to 1000 m.
22

CONFIDENTIAL
Confocal Hole and Fluorescence rejection
5 000
Spectra taken with varying confocal hole
Overall intensity diameter from 500 to 100 m at 50 m
4 000
increments
Intensity (cnt/sec)
Spectrum
3 000
2 000
1 000
0
500 1 000 1 500 2 000 2 500
Raman Shift (cm-1 )
5 000 5 000
Fluorescence Raman
4 000 4 000
Intensity (cnt/sec)
Intensity (cnt/sec)
3 000 3 000
2 000 2 000
1 000 1 000
0 0
500 1 000 1 500 2 000 2 500 500 1 000 1 500 2 000 2 500
Raman Shift (cm-1 ) Raman Shift (cm-1 )
23

CONFIDENTIAL
Confocal Hole and Fluorescence rejection
0.24 X axis: Confocal hole diameter from 500

0.22 100 m at 50 m step.
Y-axis
Spectrum
0.20
Intensity (cnt/sec)
0.18 Blue: Overall intensity (I vs. C)

Green: Fluorescence intensity (F vs. C)
0.16
Red: Raman intensity (R vs. C)
0.14
Cyan: Ratio of fluorescence intensity to
0.12 Raman intensity (R/F vs. C)
0.10 Note that the intensity ratio of Raman to
500 400 300
Hole (m)
200 100 Fluorescence ratio grows as confocal
hole diameter decreases
Let's say that the total intensity, I, is the sum of fluorescence

background, F, and Raman signal, R.
I decreases as confocal hole diameter, C, decreases (blue).
Therefore, F and R decrease as C decreases (green and red,
respectively). However, the ratio R/F increases as C decreases
(cyan).
Too small C results in too small I and thus R. Too large C results in
too small R/F. The best C is somewhere in between.
24

CONFIDENTIAL
Spectrum Slit
Slit
This is the entrance slit to spectrometer and one of the parameters
that determine spectral resolution. We recommend to relatively
narrow slit width (< 150 m) to maintain as high spectral resolution
as the system configuration allows
25

CONFIDENTIAL
Spectrum Exposure time and Accumulation
Exposure time and Exposure time for

accumulation for spectrum spectrum real time display
acquisition (RTD)
26

CONFIDENTIAL
Exposure time and Accumulation
Increasing exposure time achieves high/required S/N.

Long exposure time reduces high frequency noise and
Spectrum
increases signal, but increase the background signal,

too.
27

CONFIDENTIAL
Spectrum Real Time Display (RTD)
Spectrum RTD
Opens Spectrum window and
continuously displays
spectrum at the current
spectrometer position
Spectrometer
The spectral position at the center
over spectral range coverage
Spectrum RTD exposure time

Set separately from Acquisition
exposure time and can be
changed on the fly
28

CONFIDENTIAL
Real Time Display (RTD)
Continuously capture and display the spectrum at the

current spectrometer position
Spectrum
Neutral density filter, confocal hole diameter, slit width

and RTD exposure time can be changed on the fly
Used to fine tune the spectrum acquisition conditions
29

CONFIDENTIAL
Spectrum Extended Range
Extended Range
Opens Extended Range
window to setup the spectral
range(s) for the Spectrum
Acquisition
Indicator for Multi window

and Auto scanning at the
bottom of the window
30

CONFIDENTIAL
Extended Range
Single window
Records the spectrum with the grating stationary
Spectrum
The spectral coverage is determined by the laser wavelength and

grating
Multi window and Auto scanning
Records up to five spectral ranges. When selected, the indicator
appears at the bottom of the LS window
For each range, if needed, multiple segments are recorded with rotating
grating, and combined to produce the spectrum of the specified spectral
range
Multi window
Grating is rotated to the center of each segment
The signal from each spectral point is averaged from a single CCD pixel
Changing the number of accumulations does not change the number of
segments
Auto scanning
Grating is rotated in increments, which is determined by the number of
accumulations
The signal from each spectral point is averaged from multiple CCD pixels
Changing the number of accumulations change the number of segments
31

CONFIDENTIAL
Multi Window
360 360
340
Exposure time: 1s 340
Exposure time: 1s
Number of accumulation: 1 Number of accumulation: 2
320 320
300 300
Spectrum
280 280
260 260
240 240
Intensity (cnt/sec)
Intensity (cnt/sec)
220 220
200 200
180 180
160 160
140 140
120 120
100 100
80 80
60 60
40 40
200 400 600 800 1 000 1 200 1 400 1 600 1 800 200 400 600 800 1 000 1 200 1 400 1 600 1 800
Raman Shift (cm-1) Raman Shift (cm-1)
360 360
340
Exposure time: 1s
320 320
300 Number of accumulation: 3 300 Number of accumulation: 4

280 280
260 260
240 240
Intensity (cnt/sec)
Intensity (cnt/sec)
220 220
200 200
180 180
160 160
140 140
120 120
100 100
80 80
60 60
40 40
200 400 600 800 1 000 1 200 1 400 1 600 1 800 200 400 600 800 1 000 1 200 1 400 1 600 1 800
32

CONFIDENTIAL
Auto Scanning
360 360
340
Exposure time: 1s
Number of accumulation: 1 Number of accumulation: 2
320 320
300 300
Four segments identical to Multi Window Four segments identical to Multi Window
Spectrum
280 280
260 260
240 240
Intensity (cnt/sec)
Intensity (cnt/sec)
220 220
200 200
180 180
160 160
140 140
120 120
100 100
80 80
60 60
40 40
200 400 600 800 1 000 1 200 1 400 1 600 1 800 200 400 600 800 1 000 1 200 1 400 1 600 1 800
400 400
380 380
360
Exposure time: 1s
340 340
320 Number of accumulation: 3 320 Number of accumulation: 4

300
280
Eight segments 300
280
Thirteen segments
260 260
Intensity (cnt/sec)
Intensity (cnt/sec)
240 240
220 220
200 200
180 180
160 160
140 140
120 120
100 100
80 80
60 60
40 40
-200 0 200 400 600 800 1 000 1 200 1 400 1 600 1 800 2 000 -200 0 200 400 600 800 1 000 1 200 1 400 1 600 1 800 2 000
33

CONFIDENTIAL
Acquisition Options
From menu, select Acquisition
and then Options
Spectrum
EXPOSURE TIME,
ACCUMULATION NUMBER and
DATA NAME can be set here as
well as from the bottom tool bar
REFRESH TIME refreshes real
time when set to 0
ACCUMULATION MODE:
Average, Sum, Detector
SPIKE FILTER: None, Multi (auto
add), Multi, Single
SCANNING: Stage for point
mapping mode and Scanner for
line scanning mode
SHUTTER MODE: Auto, Open
and Close
34

CONFIDENTIAL
Experiment Setup and Data Acquisition Acquisition
Spectrum acquisition
Opens Spectrum window and start acquiring a spectrum according
to the exposure time and accumulation
35

CONFIDENTIAL
Map Mapping Area and Shape
Map shapes available for

lateral maps: lines, rectangle,
oblong and polygon
Left mouse click to select the
first point. Then drag and
drop.
36

CONFIDENTIAL
Map Mapping Properties
Mapping properties
Opens Mapping properties window
Array Size and Increment must be defined here.
37

CONFIDENTIAL
Increments
A coarse grid map (low sampling frequency) serves as a road map for the
sample.
Quick measurement over a large area
Map
Identifying major chemical components; how many, what and where.

Identifying the region of interest
A fine grid map (high sampling frequency) provides a complete picture of
the sample.
Customized measurements over a region of interest
Identifying major AND minor chemical components; how many, what and where
Morphological analysis; shape, size and locations of chemical components.
Oversampling
For samples that are near at the limit of spatial resolution, over-sampling is
recommended.
Theoretically, the spatial resolution limits the highest effective sampling
frequency or step size, which is the half of the spatial resolution. This will result in
2 points for the smallest (resolvable) object.
We have experienced, however, 5, 10, 20 times over-sampling, resulting in 10,
20, 40 points for the smallest (resolvable) object enhances the Raman image
quality significantly.
38

CONFIDENTIAL
Map Increment and Array Size
When the INCREMENT box is checked, the measurements are performed at the set increment.
When the INCREMENT box is not checked, measurements are performed at the increment
calculated from FROM, TO and ARRAY SIZE.
The array size or the number of measurements, not the mapping area, is what determines
the duration of the mapping experiment and the size of the mapping data file.
39

CONFIDENTIAL
Map Batch Setup
Right click to activate popup menu

Select insert to open Devices window
Add Z for depth profile

Add Hole for scanning confocal hole diameter
40

CONFIDENTIAL
Map Batch Setup
Only checked parameters are scanned

The lowest row is scanned first and then progress upward. In this example, x is scanned first followed
by y
To change the order of scan, use UP and DOWN options from the popup menu
To remove a parameter from the window, use REMOVE option
41

CONFIDENTIAL
Map Acquisition
Map Acquisition
Opens SpIm, Map and Point windows and starts
mapping acquisition
42

CONFIDENTIAL
Windows
Video
Map
Point
A child window that
displays a point spectrum
Map
A child window that displays
SpIm integrated intensity maps
The parent window for
map data
43

CONFIDENTIAL
Stop Stop
Stop
Either data measurement or processing, any on-going
procedure turns the Stop icon active in red. Clicking
Stop icon will stop the procedure and let the partial
result stands.
44

CONFIDENTIAL
Data Treatment Baseline Correction
Fit: fit baseline using current

OPTIONS
Sub: subtract fitted baseline
Clear: clear fitted baseline
Auto: fit, subtract and clear
Interactive Add and Remove Convert: convert the spectrum to the
baseline fitting points fitted baseline
Copy/Paste: copy and paste fitted
baseline
45

CONFIDENTIAL
Baseline Correction
Fitting points are manually added

Data Treatment
or removed as needed
Baseline fitted Baseline subtracted
46

CONFIDENTIAL
Data Treatment Spectral Normalization and Correction
Normalize: Normalizes the spectrum to the unit area

(100)
Zero: Subtract the minimum intensity from the spectrum
Subtract: Subtracts the corrector spectrum from the
selected spectrum
Multiply: Multiplies the corrector spectrum and the
selected spectrum
Correct: Subtracts the corrector spectrum the selected
spectrum after intensity adjustment
Get: Gets the current as the corrector spectrum
Delete: Removes the corrector spectrum.
Thr: Reset spectra whose maximum intensity is less than

threshold fraction (%) of maximum intensity of the map to
all zero 47

CONFIDENTIAL
Data Treatment Spectral Normalization and Correction
Zero corrected Zero corrected and normalized
48

CONFIDENTIAL
Data Treatment Smoothing
Smooth, Der1 and Der 2 use Savitsky-Golay algorithm to smooth,

take 1st derivative of and take 2nd derivative of the spectrum.
DEGREE specifies the order of polynomial and SIZE the number of
points
Median uses median filter to smooth the spectrum
Despike removes spikes (cosmic lay) whose intensity is stronger
than HEIGHT
49

CONFIDENTIAL
Data Treatment Fourier Transform
This is another smoothing tool. When a spectrum is

Fourier transformed, the low frequency data (spectral
Low High features) and high frequency data (noise) are
frequency frequency separated by eliminating the high frequency region in
the frequency domain, and then inverse Fourier
transforming, produces a spectrum where the high
frequency noise is removed.
50

CONFIDENTIAL
Fourier Transform
LIMIT: Position of the cut off point in %. This
parameter defines the smoothing factor. Click
Data Treatment
on the Go button to apply the value or drag the

cursor (red line) to select the LIMIT
interactively.
0: Fully smoothed no features
100: No smoothing
APOD: Type of the apodization function with
apodization value reaching zero at a Limit
point.
None: No apodization function
Line: Linear function
Sqr: Parabolic function
Cos: Cosine function
FILTER: Select here the type of the filter
function
None: No filter function
Traffic: Traffic function
51

CONFIDENTIAL
Data Treatment Math
CONST+ (add/subtract) and

CONST* (multiply/divide) are
binary operations between the
selected spectrum and a
constant.
These operations can be
performed interactively using
icons on the left tool bar.
52

CONFIDENTIAL
Math
FUNC 1 and FUNC 2 performs basic binary
operations (+, -, *, /), negating, power, log, exp, sin,
Data Treatment
cos, asin, acos, atan, abs, sqrt, step.

When x and y are used, they refer to individual
spectra. When a, b, c, d, are used, they refer to
axis values
COMBINE combines segments of spectra into one
full range of spectrum
Go: Perform the operation
53

CONFIDENTIAL
Data Treatment Peak Searching and Fitting
Search: find peaks to fit

automatically
Approx: approximate the initial
conditions (peak position, height
and width) for fitting
Fit: start fitting
Clear: clear all peaks
Convert: convert the spectrum
with the fitted curves
These icons allow interactively
adding and removing peaks, and
adjusting peak shapes
54

CONFIDENTIAL
Peak Searching and Fitting
FUNCTION lists fitting functions. Default fitting functions are
Gaussian, Lorentzian and Gaussian+Lorentzian combination
Data Treatment
functions.
Fitting iteration starts by clicking Fit button, and stops when
either the number of ITERATION is reached or ERROR is not
changing.
When the fitting is completed, the results are reported in Peaks
window, which opens by clicking Peaks button.
Right click to activate the popup menu for adding or removing

peaks. The table can be copied and pasted into a spreadsheet
using Copy and Paste buttons.
55

CONFIDENTIAL
When Peak Fitting has been applied to the SpIm window, the
results peak positions, peak heights, band widths, peak areas
can be mapped.
Click Peak button from Peaks window and check MAP box of
the parameter to be mapped. Click OK to open Param window
56

CONFIDENTIAL
Variables opens Variables window to set parameters for fitting

functions.
Enter user defined functions at Functions.

57

CONFIDENTIAL
Options: customize rendering
58

CONFIDENTIAL
Data Treatment Profile from Image
Profile window displays vertical or

horizontal intensity profile along a line in
an image
59

CONFIDENTIAL
Data Treatment Profile from Image
VER shows the intensity profile along the vertical

line.
HOR shows the intensity profile along the horizontal
line.
60

CONFIDENTIAL
Data Treatment Map Analysis
When USE is checked,

intensity between FROM and
TO is mapped in the color
designated. BASELINE
subtracts a linear baseline
between FROM and TO.
Intensity ratio of GREEN over

BLUE is mapped in cyan.
Check SPECTRUM box to see After applying data treatment

the spectrum of the selected to Point window, click on the
data point(s) Correct button to update
61 SpIm.

CONFIDENTIAL
Interactively adjust the spectral range for

intensity mapping red for red image,
green for green image and blue for blue
image
62

CONFIDENTIAL
Spectral ranges for intensity maps can be set in three places Point window, SpIm
window and Map Analysis window. All adjustments are synchronized.
63

CONFIDENTIAL
Intensity maps in Map window.
64

CONFIDENTIAL
Data Treatment Create Spectral Profile
Separately recorded spectra can be grouped into a map

from Spectral Profile window.
Input NAME, select the first spectrum in the series, and
click New. This creates a map file and opens map
windows SpIm, Point and Map. Select the next
spectrum and click Add or Insert till all spectra are added.
To remove a spectrum from the map, select the spectrum
and click Delete.
65

CONFIDENTIAL
Data Treatment Modeling
When a meaningful spectrum is available, it can be

used as a model.
Select the spectrum from Spectrum, Point or Model
window.
Click Get button. The selected spectrum is placed in
the Model window and the corresponding scores image
is created in matching color.
66

CONFIDENTIAL
Data Treatment Modeling
When no meaningful spectrum is available, or the

sample is totally unknown, Modeling can calculate
statistically representative models.
Determine the number of factors and click Create
button. Models and scores images are created.
67

CONFIDENTIAL
The Number of Spectra in a Raman Map
Modeling Multivariate/chemometrics analyses are statistical analysis
methods. Therefore, in a Raman map, if there are multiple spectra
of the same chemical compound, even though the S/N of individual
Map
spectra is somewhat poor, the repetition can contribute to recover

the S/N statistically, similarly to averaging multiple accumulations in
a single point measurement.
In LS, the number of spectra in a map is determined by the array
size. Increase the array size increase the number of spectra in
the map increase the chance to increase the number of spectra
of the same chemical increase the chance to increase the S/N of
the model created in Modeling without reference spectra.
When reference spectra is applied, the statistical nature of
multivarate/chemoemtrics is irrelevant).
68

CONFIDENTIAL
Modeling and the Number of Spectra in a Raman
Map
When applied without reference spectra, Modeling is a statistical
analysis method. Therefore, in a Raman map, if there are multiple
spectra of the same chemical compound, even though the S/N of
Map
individual spectra is somewhat poor, the repetition can contribute to

recover the S/N statistically, similarly to averaging multiple
accumulations in a single point measurement.
The number of spectra in a map is determined by the array size.
Increase the array size increase the number of spectra in the
map increase the chance to increase the number of spectra of
the same chemical increase the chance to increase the S/N of
the model created in Modeling without reference spectra.
When reference spectra are used, the statistical nature of Modeling
is irrelevant, and the number of spectra has no affect on Models
S/N.
69

CONFIDENTIAL
Imposition
Select options for imposition
Visualization
Right click in
Video window Select a window with
2D image (Map, Scores
or Param) to overlay
70

CONFIDENTIAL
Popup Menus
Video window Map window Score window
Visualization
Spectrum window Point windows Model windows
Activated by clicking right mouse button in the window. These menus are window type sensitive
71

CONFIDENTIAL
Popup Menus
SpIm window in 1D display SpIm in 3D display
Visualization
1.0
Intensity (cnt/sec)
0.8
1.0
0.6
0.4
0.5
0.2
950
0.0 Ram
an S 1 000 10
hift ( m)
960 980 1 000
-1
1 020 1 040
1/cm X (
Raman Shift (cm ) )
SpIm in 2D display
7.0
8.0
9.0
X (m)
10.0
11.0
12.0 5 1/cm
13.0
950 1 000
Raman Shift (cm-1 )
Activated by clicking right mouse button in the window. These menus are window type sensitive
72

CONFIDENTIAL
Format and Scale
MODE:
Single displays one data set at a time
Visualization
Overlay overlays multiple data sets on top of each other

Stack stacks multiple spectra with a constant offset
Tile each data set is displayed in a sub window
SCALE:
FREEZE fixes FROM and TO
AUTO automatically rescales to whatever changes made
REL uses separate axis for each of multiple data sets
Single display mode
Multi display (Overlay, Stack or Tile) mode
Switch between spectra
73

CONFIDENTIAL
Format and Scale
Overlay Stack
Visualization
Tile
74

CONFIDENTIAL
Visualization Axes
3D axes
2D axes
75

CONFIDENTIAL
Visualization Cursor
Spectrum, Point and Model windows Video, Map and Score windows
Line: single spectral position Cross: a point
Double: two spectral positions Rect: an area for averaging
Cross and Level
Peak: peak position (solid line) and
approximated FWHM (dashed lines)
76

CONFIDENTIAL
Colors and Shapes
Spectrum, Point and Model
windows: Styles, colors, line
Visualization
thickness, point sizes, etc.
Map and Score windows

colors and interpolation options
SpIm window in 3D display

colors
SpIm window in 1D display

colors and rendering options (All
spectra or Limits)
77

CONFIDENTIAL
Thank you
78


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CONFIDENTIAL

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

File I/O Data Information Experiment setup Data processing

Neutral density filter Exposure time

Spectrometer Laser block/unblock

Grating System setup

4 CCD temperature monitor

2009 HORIBA, Ltd. All rights reserved.

Right click on the background

2009 HORIBA, Ltd. All rights reserved.

Unique set of utilities on the left tool bar

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

thorough the signal for polarized

2009 HORIBA, Ltd. All rights reserved.

Rescale both X- and Y-axis Center cursors

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

Parameters window shows all

2009 HORIBA, Ltd. All rights reserved.

Opens Video window and display real-time

The last image is captured and can be saved

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

Green lasers: 500 600 nm

2009 HORIBA, Ltd. All rights reserved.

500 1000 1500 2000 2500 3000

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

A higher groove density grating provides higher spectral

2009 HORIBA, Ltd. All rights reserved.

500 Lidocane, 1800 gr/mm

500 1 000 1 500 2 000 2 500 3 000 3 500

Lidocane, 300 gr/mm

500 1 000 1 500 2 000 2 500 3 000 3 500

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

The main role of the confocal hole is to reject out-of-

The main role of the slit is to reduce the size of image at

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

0.24 X axis: Confocal hole diameter from 500

0.18 Blue: Overall intensity (I vs. C)

Let's say that the total intensity, I, is the sum of fluorescence

2009 HORIBA, Ltd. All rights reserved.

2009 HORIBA, Ltd. All rights reserved.

Exposure time and Exposure time for

2009 HORIBA, Ltd. All rights reserved.

Increasing exposure time achieves high/required S/N.

increases signal, but increase the background signal,

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Spectrum RTD exposure time

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Continuously capture and display the spectrum at the

Neutral density filter, confocal hole diameter, slit width

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Indicator for Multi window

2009 HORIBA, Ltd. All rights reserved.

The spectral coverage is determined by the laser wavelength and

2009 HORIBA, Ltd. All rights reserved.