Robust and rapidly tunable light source for
SRS/CARS microscopy with low-intensity noise
Heiko Linnenbank,
a,b,
* Tobias Steinle,
a,b
Florian Mörz,
a,b
Moritz Flöss,
b
Han Cui,
c
Andrew Glidle,
c
and
Harald Giessen
a,b
a
SI Stuttgart Instruments GmbH, Öhringen, Germany
b
University of Stuttgart, 4th Physics Institute and Research Center SCoPE, Stuttgart, Germany
c
University of Glasgow, College of Science and Engineering, Division of Biomedical Engineering, Glasgow, United Kingdom
Abstract. We present a fully automated laser system with low-intensity noise for coherent Raman scattering
microscopy. The robust two-color system is pumped by a solid-state oscillator, which provides Stokes pulses
fixed at 1043 nm. The tunable pump pulses of 750 to 950 nm are generated by a frequency-doubled fiber-
feedback femtosecond optical parametric oscillator. The resulting pulse duration of 1.2 ps provides a viable
compromise between optimal coherent Raman scattering signal and the necessary spectral resolution. Thus
a spectral range of 1015 to 3695 cm
−1
with spectral resolution of <13 cm
−1
can be addressed.
Keywords: label-free imaging; stimulated Raman scattering; optical parametric amplifier; spectral compression.
Received Jul. 4, 2019; accepted for publication Aug. 28, 2019; published online Sep. 24, 2019.
© The Authors. Published by SPIE and CLP under a Creative Commons Attribution 4.0 Unported License. Distribution or
reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
[DOI: 10.1117/1.AP.1.5.055001]
1 Introduction
Coherent Raman scattering (CRS) imaging is based on a multi-
photon scatteri ng process that employs two near-infrared laser
pulses to excite Raman modes in the midinfrared spectral range.
Owing to its chemical selectivity without labeling, it has found a
wide scope of applications in biomedical microscopy, such as
live cell, tissue, or DNA imaging,
1–5
over the past years. Its most
prominent representatives are coherent anti-Stokes Raman scat-
tering (CARS) and sti mulated Raman scattering (SRS).
6
In the
case of CARS, two beams, the so-called pump and Stokes beam,
with an energy difference matching the Raman resonance Ω,
interact according to a four-wave-mixing process. Thus coher-
ent radiation at the anti-Stokes frequency ω
CARS
¼ 2ω
P
− ω
S
is
generated, as depicted in Fig. 1. In contrast to CARS, SRS is an
inelastic scattering proces s, where energy is transferred from the
pump to the Stokes beam, thus either the change in pump
[stimulated Raman loss (SRL)] or Stokes intensity [stimulated
Raman gain (SRG)] can be detected. SRS, unlike spontaneous
spectroscopy and CARS,
7
is therefore free from fluorescent and
nonresonant background. In addition, SRS provides quantitative
information about the chemical constituents, due to its linear
signal scaling.
8–10
Although in the case of CARS a very weak but spectrally
easy to separate third beam is generated, a tiny change of either
the pump or the Stokes beam has to be measured for SRS. Thus
lock-in-based detection is required for the latter.
1
This imp lies
that for SRS an ultralow-intensity laser noise, ideally at the
shot-noise limit, is required. Simultaneously, excellent long-
term average power stability and reproducibility are required
for laser scanning imaging, and at least one color must be rap-
idly tunable while maintaining its spatiotemporal properties.
The main regions of interest are the H-stretching vibrational
region (2500 to 3500 cm
−1
) as well as the fingerprint region
(500 to 1700 cm
−1
). In order to face real-world applications,
the source is required to be cost-effective and easy to use, and
thus to be fully automated.
The vast majority of work in CRS in the past decade was
based on picosecond, green-pumped optical parametric oscilla-
tors (OPOs),
8,11
as well as electronicall y synchronized Ti:sap-
phire laser systems.
9
Recent demonstrations of novel CRS
sources include Fourier-transform spectral retrieval,
12
chirped
pulse spectral focusing,
13,14
as well as photonic time stretch,
15
ps-laser diodes,
16
and swe pt-source SRS.
17
These techniques are
able to achieve higher information volume
6
but to date are still
quite complex to operate and exhibit alignment sensitivity of the
detection system.
*Address all correspondence to Heiko Linnenbank, E-mail: h.linnenbank@pi4.uni-
stuttgart.de
Letter
Advanced Photonics 055001-1 Sep∕Oct 2019
•
Vol. 1(5)