Semrock Master Catalog 2018

Multiphoton Filters Common Specifications
Common Specifications
Property Emitter LWP Dichroic Comment
Passband
Transmission
Guaranteed > 90% > 93% Averaged over any 50 nm (emitter) or 10 nm
Typical > 95% > 95%
(dichroic) window within the passband. For SWP
dichroic specifications, see page 44 .
Dichroic Reflection LWP N/A > 98% Averaged over any 30 nm window within the reflection
band. For SWP dichroic specifications, see page 44.
Autofluorescence Ultra-low Ultra-low Fused silica substrate
Blocking
Pulse Dispersion
Emitter Orientation
Dichroic Orientation
Emitter filters have exceptional blocking over the Ti:Sapphire laser range as needed to achieve superb
signal-to-noise ratios even when using an extended-response PMT or a CCD camera or other siliconbased
detector.
LWP dichroic beamsplitters are suitable for use with 100 femtosecond Gaussian laser pulses. For SWP
dichroic beamsplitters, see Group Delay Dispersion and Polarization Technical Note at
www.semrock.com
The emitter orientation does not affect its performance; therefore there is no arrow on the ring to
denote a preferred orientation.
For the LWP dichroic, the reflective coating side should face toward detector and sample. For the SWP
dichroic, the reflective coating side should face towards laser as shown in the diagram on page 27.
Fluorophores
Single-band
Sets
Multiband
Sets
Microscope Compatibility
These filters fit most standard-sized microscope cubes from Nikon, Olympus, and Zeiss and may
also be mounted in optical bench mounts. Contact Semrock for special filter sizes.
Cubes
TECHNICAL NOTE
Multiphoton Filters
In multiphoton fluorescence microscopy, fluorescent molecules that
tag targets of interest are excited and subsequently emit fluorescent
photons that are collected to form an image. However, in a two-photon
microscope, the molecule is not excited with a single photon as it is in
traditional fluorescence microscopy, but instead, two photons, each
with twice the wavelength, are absorbed simultaneously to excite
the molecule.
Ti:Sapphire
Laser
Source
Laser Beam
Scan Head
Dichroic Beamsplitter
Emitter Filter
Figure 1: Typical
configuration of
a multiphoton
fluorescence
microscope
Laser
Sets
NLO
Filters
As shown in Figure 1, a typical system is comprised of an excitation
laser, scanning and imaging optics, a sensitive detector (usually a
photomultiplier tube), and optical filters for separating the fluorescence
from the laser (dichroic beamsplitter) and blocking the laser light from
the detector (emission filter).
The advantages offered by multiphoton imaging systems include: true
three-dimensional imaging like confocal microscopy; the ability to image
deep inside of live tissue; elimination of out-of-plane fluorescence; and
reduction of photobleaching away from the focal plane to increase
sample longevity. Now Semrock has brought enhanced performance
to multiphoton users by introducing optical filters with ultra-high
transmission in the passbands, steep transitions, and guaranteed deep
blocking everywhere it is needed. Given how much investment is
typically required for the excitation laser and other complex elements
of multiphoton imaging systems, these filters represent a simple and
inexpensive upgrade to substantially boost system performance.
Depth 100 µm
Sample
Detector
x-z projection x-y images y-z projection
1
2
Individual
Filters
Dichroic
Beamsplitters
Exciting research using Semrock multiphoton filters demonstrates the power of
fluorescent Ca 2+ indicator proteins (FCIPs) for studying Ca 2+ dynamics in live cells using
two-photon microscopy. Three-dimensional reconstructions of a layer 2/3 neuron
expressing a fluorescent protein CerTN-L15. Middle: 3 selected images (each taken
at depth marked by respective number on the left and right).
Image courtesy of Prof. Dr. Olga Garaschuk of the Institute of Neuroscience at
the Technical University of Munich. (Modified from Heim et al., Nat. Methods,
4(2): 127-9, Feb. 2007).
1
2
3
Depth 260 µm
3
1
2
3
Tunable
Filters
47
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