TriM Scope II Series

Components

Lasers

CRONUS laser for 2-photon microscopy

The CRONUS laser is a novel, femtosecond pulse laser expanding the possibilities of 2-photon microscopy. It provides two tunable (680–960 nm and 960–1300 nm) and one fixed (1025 nm) independent but synchronized laser beams. Its gap-free, broad tuning curves cover the entire excitation spectrum relevant for 2-photon applications. The CRONUS laser thus allows a free choice of fluorophores that can be excited simultaneously and moreover the detection of non-linear optical processes, such as second and third harmonic generation (SHG and THG).

Thus far, the combination of a Ti:Sapphire laser with an optical parametric oscillator (OPO) has been considered the gold standard for the simultaneous excitation of multiple fluorophores in 2-photon microscopy. However, this combination does not fully cover the excitation spectra of many fluorescent dyes and markers frequently used in life sciences. In contrast, the CRONUS laser provides a continuous excitation range between 680 and 1300 nm (see tuning curves). Single or dual output broad band lasers, which are integrated into 2-photon microscopy systems nowadays, do not have the capability to excite multiple dyes at once. Integrated into our TriM Scope systems, the CRONUS laser meets this challenge with its three synchronized output laser lines.

Benefits of the CRONUS laser for 2-photon microscopy in a nutshell:

  • Free choice of fluorophores for 2-photon microscopy through two tunable (680–960 nm, 960–1300 nm) and one fixed (1025 nm) synchronized output channels
  • Simultaneous excitation of fluorescent probes, calcium indicators, or opsins right at their spectral absorption maxima
  • Compact and cost-efficient

Third-party lasers for 2-photon microscopes

The TriM Scope 2-photon microscopy platforms can also be equipped with a large variety of third-party lasers, either individually or in combination. These include classical Ti:Sapphire (680–1080 nm), ultra-wide tunable (660–1320 nm), OPO (1010–1340 nm), and fixed-wavelength femtosecond (1040 or 1070 nm), visible lasers for phototreatment.

This range of options enables high flexibility regarding fluorophores and optogenetic actuators with different excitation ranges, as well as simultaneous excitation of several dyes.


Our optional module for dispersion compensation keeps the laser pulse length short, which guarantees efficient excitation with any type of optical layout and objective.

A common problem when using different laser wavelengths for inspection of multiple dyes or for simultaneous imaging and photomanipulation is the mismatch of focal planes. All TriM Scope systems can be equipped with adjustable telescope optics to overcome these hurdles.

Type Wavelength [nm] Models (examples) Application Probes (examples)
 Ultra-wide tunable 680-1300

CRONUS laser

Spectra Physics InSight X3, Coherent Chameleon Discovery

Fluorescence GFP, GCaMP, YFP, , RFP, mCherry
Optogenetics & Uncaging ChR2, MNI-Glu, RuBi-GABA, C1V1 
SHG, THG intrinsic
Ti:Sapphire  690-1080 Coherent Chameleon Ultra II, Spectra Physics MaiTai Fluorescence RFP, GCaMP, YFP 
Optogenetics & Uncaging ChR2, MNI-Glu, RuBi-GABA
SHG intrinsic
 OPO  1010-1340 Coherent Chameleon  MPX (pumped with Ultra II) Fluorescence  RFP, mCherry
Optogenetics  C1V1
SHG, THG  intrinsic
Fixed-Wavelength Femtosecond 1045, 1070 Coherent Fidelity-2, Spectra Physics HighQ-2 Optogenetics C1V1, ReaChR

Visible lasers (1-photon)

488, 532, 561, 594, 637, etc. Coherent Obis Fluorescence GFP, YFP, mCherry
Optogenetics & Uncaging ChR2, MNI-Glu, Halorhodopsin, Chrimson