Lightnovo miniRaman User manual

miniRaman microscope
version 1.0
English
lightnovo.com
User Manual

2
Table of Contents
Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
General Information . . . . . . . . . . . . . . . . . . . . . 4
Warning labels . . . . . . . . . . . . . . . . . . . . . . . . 5
Safety instructions . . . . . . . . . . . . . . . . . . . . . . 5
Laser Safety . . . . . . . . . . . . . . . . . . . . . . . . . 6
Installation . . . . . . . . . . . . . . . . . . . . . . . . 8
General Information . . . . . . . . . . . . . . . . . . . . . 8
Delivery Scope . . . . . . . . . . . . . . . . . . . . . . . . 8
Site requirements . . . . . . . . . . . . . . . . . . . . . . 9
Cable Connections . . . . . . . . . . . . . . . . . . . . . . 10
Software Installation . . . . . . . . . . . . . . . . . . . . . 11
Removal and installation of miniRaman spectrometer . . . . . . . . . 12
Overview . . . . . . . . . . . . . . . . . . . . . . . . . 13
General information. . . . . . . . . . . . . . . . . . . . . . 13
Specication . . . . . . . . . . . . . . . . . . . . . . . . 14
Instrument design . . . . . . . . . . . . . . . . . . . . . . 16
General overview of control elements and components . . . . . . . . 17
Operation . . . . . . . . . . . . . . . . . . . . . . . . . 18
Hardware connection . . . . . . . . . . . . . . . . . . . . . 18
Operation procedure . . . . . . . . . . . . . . . . . . . . . 19
Upright and Inverted operation . . . . . . . . . . . . . . . . . 28
Technical drawings. . . . . . . . . . . . . . . . . . . . 29
Service addresses . . . . . . . . . . . . . . . . . . . . 30

4
Safety
1General Information
Read the following safety instructions carefully before operating miniRaman micro-
scope and keep this manual for future reference available at any time. Always observe
the instructions described in this manual to ensure user safety and to avoid property
damage. Improper use or failure to follow these safety instructions can result in seri-
ous injuries and/or property damage. Any non-observance of the precautions will
infringe the intended use (i.e. performing measurements by Raman spectroscopy) of
miniRaman microscope. In this case Lightnovo ApS will not assume any liability. It is the
operator’s duty to plan and implement all necessary safety measures and to supervise
their observance. Moreover, the operator must ensure that miniRaman microscope is
in proper functioning condition. A safe and faultless operation can only be guaranteed
if miniRaman microscope is transported, stored, installed, operated and maintained
properly according to the procedures described in this manual. Never remove or deac-
tivate any supporting safety systems during miniRaman microscope operation. Ensure
that objects and/or material not required for the measurement is out of the instrument
operating area.
Qualied Personnel
Primary installation and all maintenance and repair works not described in this manual
should only be performed by Lightnovo service personnel. Only authorized operat-
ing personnel that have been briefed about the instrument operation and all relevant
safety aspects should operate and maintain (i.e. only maintenance works that are de-
scribed in this manual) the instrument. All repairs, adjustments and alignments on any
miniRaman microscope component must be performed in accordance with the safety
regulations and standards applied in the country in which the instrument is installed.
Correct Usage
miniRaman microscope and its components should only be used according to the
instructions described in the manual or advised by a Lightnovo engineer. In case of
accessories or components made by other manufacturers and used in connection with
the microscope, Lightnovo does not assume any liability for safe operation and proper
functioning.

5
Safety
2
3
Warning labels
Safety instructions
This warning symbol indicates the existence of laser radia-
tion. A Class 3R laser is considered safe if handled carefully,
with restricted beam viewing. With a class 3R laser, the maxi-
mum permissible exposure (MPE) can be exceeded, but with
a low risk of injury. Visible continuous lasers in Class 3R are limited to 5 milliwatts.
This warning symbol indicates the existence of laser radia-
tion. Class 3B lasers are hazardous for eye exposure. They
can heat skin and materials but are not considered a burn
hazard. For visible-light lasers, Class 3B lasers’ output power
is between 5 and 499 milliwatts. Class 3B lasers are normally
hazardous under direct beam viewing conditions, but are normally safe when viewing
diuse reections.
The following chapters describe all relevant safety aspects of the instrument operation.
Depending on the degree of hazard the safety instructions are classied as follows:
Danger
indicates that death, severe personal injury or substantial property damage WILL result
if proper precautions are not taken.
Warning
indicates that death, severe personal injury or substantial property damage CAN result
if proper precautions are not taken.
Caution
indicates that minor personal injury or property damage CAN result if proper precau-
tions are not taken. Important draws your attention to a particularly important infor-
mation.
Note
draws your attention to an useful information on the product, e.g. product operation
or to a special part of the manual.
The safety instructions Danger, Warning and Caution are marked by the corresponding
warning labels.

6
4Laser Safety
General Information
The analysis system miniRaman microscope uses the light of a lasers (660 and 785nm)
that are installed in miniRaman spectrometer. The used laser diodes emit visible and
partially visible laser radiation in the near infrared region.
According to the standard EN 60825-1/10.2003, model MiniRaman microscope SERS
is laser class 3R product and it is considered safe if handled carefully, with restricted
beam viewing.
According to the standard EN 60825-1/10.2003, miniRaman microscope models:
MiniRaman microscope Standard, MiniRaman microscope Standard Dual, MiniRaman
microscope Power Dual, MiniRaman microscope Power are Class 3B lasers products.
Therefore, they are normally hazardous under direct beam viewing conditions, but are
normally safe when viewing diuse reections.
Safety Features
The laser protective enclosure shields the area around the sample stage. The enclosure
is opened and closed manually. For safety reasons, the operator has to close the laser
protective enclosure before starting measurement. This safety precaution protects the
operator against accidental exposure to hazardous laser radiation exiting the objective.
Figure 1. Open Laser Protective Enclosure Closed Laser Protective Enclosure
Safety

7
Safety Instructions
In additions to the safety instructions given below, also comply with all local regulations
concerning laser safety.
The analysis system is specied as a laser class 3R/3B product (depending on the
model, see overview section), i.e. it considered safe if handled carefully, with restricted
beam viewing. Nonetheless, observe the following safety instructions:
Warning:
Avoid eye and skin exposure to direct or scattered laser radiation under
all circumstances! Failure to do so can cause permanent and irreversible
eye damage and/or serious skin injuries!
Do NOT remove the laser protective enclosure!
Safety

8
Installation
1
2
General Information
Delivery Scope
Unpacking and initial installation including hardware setup and cable connection is
done by qualied Lightnovo service personnel. The operating company has to provide
the required utilities and an installation site that meets the site requirements described
in this chapter.
Standard Components
miniRaman microscope (including user manual and quality test report)
Several cables: power cord, USB cable. (See section Cable Connections below in this
chapter.)
Accessories (includes spares, adaptors, objective lenses, sample preparation tools etc.)
Inspecting the Packaging
After having received miniRaman microscope, inspect the packaging for damages. If
there are any signs of damage, contact your local shipping representative before open-
ing the shipping box.
Warning:
Do not put miniRaman microscope into operation if there are signs of damage. Failure
to do so may result in severe personal injuries and/or property damage.
Transportation
When transporting
the microscope, use
the original case to
avoid damages.
Figure 2. Transportation
case for miniRaman micro-
scope

9
Installation
3Site requirements
Space Requirements
miniRaman microscope requires a space of 17cm in diameter and 28cm in height. (For
the exact instrument dimensions refer to Specication.) At the rear instrument side,
take a clearance of at least 5cm into account.
When dening the installation site, take into consideration that the power supply con-
nections are easily accessible at any time.
Environmental Requirements
To ensure optimum instrument performance and long-term reliability the following
environmental conditions are essential:
• Ambient temperature range:18 - 35°C (64 - 95°F)
• Humidity (non-condensing): ≤ 70% (relative humidity)
Temperature variations can impair the results of long-term measurements. Therefore,
the temperature variations should be less than 1°C an hour and not more than 2°C per
day for high spatial resolution type of measurements.
Vibration
Ideally, the instrument is not installed near vibration sources (e.g. ventilation hoods, air
conditioners, motors, elevator).
Power Supply
The instrument power supply unit automatically adapts to the most common power
sources.
Valid voltage range:
• 100V microscope system: 100V to 120V AC 10%
• 200V microscope system: 220V to 240V AC 10%
• Valid frequency range: 50/60Hz
MiniRaman microscope is an instrument of protection class I (electric safety).
Note:
A power outlet box to which miniRaman microscope connected provides a suitable solution.
miniRaman microscope requires a stable and horizontal base which can carry the instru-
ment (ca. 7kg) and possible accessories. This is critical for applications that require high
spatial resolution.
Note:
For some kinds of application, the opportunity to black out the room is advantageous

10
Installation
4Cable Connections
Caution:
To avoid personal injury and spectrometer damage, connect miniRa-
man microscope only to a socket outlet with earthing contact.
Please unpack miniRamaman
microscope and remove it from the
transportation case (see Figure 3).
Please remove power adaptor
(Figure4) and connect it to the right
connector on the rear panel of the
miniRaman microscope (Figure 5).
Then connect USB cable to the left
connector on the rear panel and
connect it to PC from the other side.
Figure 3. miniRaman microscope with transportation case.
Figure 4. Power adaptor.

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Installation
Figure 5. Rear side – overview of the connector
sockets (left connector is USB 2.0 for connection
with PC via USB cable, right connector is for power
supply 12V).
5Software Installation
1. Download miniRaman microscope software package from Lightnovo website:
https://lightnovo.com/lightnovo-software/
Please select version 32 or 64 bit depending on your operation system.
2. Install all drivers from the corresponding software folder.
3. Run Miraspec.exe le to start the data acquisition software.
4. Software is ready for operation.

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6Removal and installation of miniRaman spectrometer
MiniRaman microscope has an in-build product inside which is miniRaman spectrom-
eter. It can be inserted and removed by the user without the need of realignment.
Removal steps are shown below.
1
4
7
2
5
8
3
6
9
remove top cover
unscrew three bolts on the rear panel
remove small magnet
unscrew four side bolts of rear panel
unscrew three bolts that hold
miniRaman spectrometer
assembly laser safety kit
remove rear slide cover
remove miniRaman spectrometer
miniRaman spectrometer is ready to use
gure 6. Removal steps of the miniRaman spectrometer from the miniRaman microscope

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Overview
1General information
MiniRaman microscope has a modular design with reation and o-axis white light
illumination microscopy unit combined with motorized 3D stage and miniRaman spec-
trometer.
(See section Instrument Design below in this chapter.)
This instrumental setup allows for both sample viewing using the optical microscopy
capabilities and performing measurements by Raman spectroscopy simultaneously.
The visual observation reveals morphological details of a sample (e.g. color, size,
shape), whereas the spectroscopic measurement reveals information about the mo-
lecular structure and chemical composition of a sample.
This instrumental setup is designed for demanding R&D application in materials sci-
ence, pharmacy, life science or forensics, for example. Possible elds of application are
analyses of SERS signals on plasmonic substrates, thin sample lms, nano particles and
surfaces.
MiniRaman microscope is also suitable for samples that tend to uoresce when ex-
posed to laser radiation. Due to the usage of a 785nm laser, the excitation energy is low
enough for these samples not to uoresces or only to a minor degree.
Note:
In Raman spectroscopy, sample uorescence can yield a much more intense signal than the
Raman scatter of the sample, masking any Raman bands in the spectrum. Therefore, Raman
spectroscopy is normally not a suitable analysis technique for uorescent samples.

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Overview
2Specication
Lasers
Depending on the selection of the miniRaman spectrometer model miniRaman micro-
scope could be congured in the following models with dierent laser parameters:
• MiniRaman microscope Standard (includes one laser with optical isolation: 785
nm, power range on a sample 5-50 mW)
• MiniRaman microscope Power (includes one laser with optical isolation: 785 nm,
power range on a sample 15-150 mW)
• MiniRaman microscope SERS (includes one laser with optical isolation: 785 nm,
power range on a sample 0.5-5 mW)
• MiniRaman microscope Power Dual (includes two lasers with optical isolation:
785 nm, power range on a sample 15-150 mW; 660 nm, power range on a sample 1-32
mW)
• MiniRaman microscope Standard Dual (includes two lasers with optical isola-
tion: 785 nm, power range on a sample 5-50 mW; 660 nm, power range on a sample
1-32 mW)
Spectral range
• 400-2700 cm-1 (at 785 nm laser excitation),
• 2750-4500 cm-1 (at 660 nm laser excitation)
Spectral resolution
• 10-15 cm-1 (slit size dependent; slit size can be customized)
Sensitivity in point mode at laser wavelength 785 nm (determined as SNR of poly-
styrene spectrum)
• SNR 350:1
• spectral range 400-2700 cm-1
• laser wavelength: 785 nm
• laser power: 100 mW
• integration time: 0.1s
• number of repetitions: 1
Sensitivity in point mode from laser wavelength 660 nm (determined as SNR of
polystyrene spectrum).
• SNR 150:1
• spectral range 2750-4500 cm-1
• laser wavelength: 660 nm
• laser power: 30 mW
• integration time: 0.5 s
• number of repetitions: 1

15
Overview
Spatial resolution
• Diraction limited (microscope objective dependent)
• 600 nm (lateral) at 660 nm excitation at NA=0.95 (100x)
• 750 nm (lateral) at 785 nm excitation at NA=0.95 (100x)
White light illumination
• Reected light microscopy with simultaneous laser spot visualization and Raman
spectra registration
• Transmittance light microscopy with simultaneous laser spot visualization and
Raman spectra registration
• O-axis illumination microscopy with simultaneous laser spot visualization and
Raman spectra registration
Microscopy conguration mode
• up-right
• inverted
Motorized stage
• lateral step size 50nm, axial step size 50nm
• bi-directional reproducibility 0.5um
• closed loop
• travel range in XYZ is 26mm
Weight
7kg
Dimensions
12x13x28 cm

16
Overview
2Instrument design
MiniRaman microscope consists of three key units (see Figure 7):
1. miniRaman spectrometer
2. white light microscopy unit
3. motorized 3D stage
MiniRaman spectrometer can be installed and taken out by the user if necessary as
shown in Figure 6.
Key system components are shown on Figure 7.
Figure 7. Opto-mechanical design of the miniRaman microscope.
Motorized 3D stage
Sample holder
Sample clamp
Microscope objective
miniRaman spectrometer
LED for o-axis white
light microscopy
LED for o-axis white
light microscopy
LED for reected white
light microscopy
CMOS sensor

17
3General overview of control elements and components
Overview
MiniRaman microscope has 6 control elements:
1. Laser with wavelength 785nm (ON/OFF and power control)
2. Laser with wavelength 660nm (ON/OFF and power control)
3. Spectroscopic sensor for Raman spectra acquisition (exposure, gain, row selec-
tion and binning control)
4. LED for reected light microscopy (power control)
5. LED for o-axis light microscopy (power control)
6. LED for transmittance light microscopy (power control)
7. CMOS sensor for white light microscopy (exposure, gain and color balance con-
trol)
8. 3D stage for sample movement and automated Raman mapping (sample scan-
ning control)

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Operation
1Hardware connection
MiniRaman microscope has two wire connections:
1. USB cable for system control and data transfer
2. Power cable (12V)
Both cables could be connected (order does not matter). User should wait for LED 1
shining “Blue” (constant) and miniRaman spectrometer bottom blinking “Blue”. Then
software is ready for connection (“Connect” bottom in Miraspec). After connection LED
2 should be “Green”. It means that device is properly connected. If LED 2 is “Red” – de-
vice has connection problems. Please try to reconnect USB cable and power cable. If it
does not help – contact service ([e-mail]).
If connection with PC is lost bottoms 2 and 3 will not shine.
During Raman mapping LED 3 will be blinking “Yellow”.
Figure 8. LED indicators of miniRaman microscope.
123
Power ON (Blue)
Connection to PC
(Green) Fault (Red)
Scanning (blinking yellow)
Location of temperature
and humidity sensor
miniRaman spectrometer
botton (used for
spectrometer reboot)

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2Operation procedure
Operation
Switching the system ON/OFF
It is highly recommended to switch o the miniRaman microscope LEDs for white light
microscopy when they are not used for a longer period of time (several hours) and
overnight. This action will prolong the service lifetime of the microscope LED.
When the analysis system is not used for a longer period of time, it is also highly rec-
ommended to switch o also the Raman excitation laser (785nm and/or 660nm). This
action will prolong the service lifetime of the laser.
Switching o the PC is facultative. Take into account that each PC reboot requires a
recalibration of the motorized stage.
Starting the Miraspec software
1. Connection
Press “Connect” bottom in Connection window (see Figure 9). This will initialize
hardware accessories. Connection window can be found in Menu/Tools/Connection
(Figure11).
Figure 9. miniRaman microscope software interface; connection of accessories

20
Operation
2. Turn ON/OFF lasers
Press “On” or “O” bottoms, see below.
Figure 11. Tools menu
Figure 10. Switch on/o laser