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FRET Microscope Filters | SIMTRUM Photonics Store

Products / Optics /Filters /Filters for FRET Microscope /
Filters for FRET Microscope
Overview Specifications Accessories Order Now!
FRET is an effective tool for measuring the distance between two different sites, as well as the change in distance, and is very sensitive to changes in distance, known as a "spectroscopic ruler", with a characteristic distance of 1 to 10 nm.
 
If there is an overlap between the emission spectra of one fluorophore (donor) and the absorption spectrum of another fluorophore (acceptor), then when the distance between the two is between 1 and 10 nm, the donor will pass through the dipole-dipole interaction (dipole-dipole interaction) transfers energy to the acceptor in a non-radiative manner, and the donor returns to the ground state without emitting fluorescence, while the acceptor is excited and emits fluorescence when decaying to the ground state, a process called fluorescence resonance energy transfer.
The FRET efficiency value can be calculated by measuring the fluorescence intensity or fluorescence lifetime as follows: E = 1 - FDA/FD or E = 1 - τDA/τD where FD is the fluorescence intensity of the donor when the receptor is absent and FDA is the fluorescence intensity of the donor when the receptor is present. τD is the fluorescence lifetime of the donor in the absence of the acceptor, and τDA is the fluorescence lifetime of the donor in the absence of the acceptor.
Depending on the experimental needs or microscope configuration, FRET filters can be configured in different combinations. In a donor-single-excitation configuration, only one excitation plate is included to excite the donor, and two emitters are included to receive fluorescence from the donor and acceptor. The two emitters need to be mounted on the filter wheel, and the different emitters need to be switched through the rotor to receive the fluorescence of the donor and acceptor respectively. If the emitter is installed in a beam splitter, an additional dichroic mirror for beam splitting is required.

Cy3 - Cy5 FRET pair,Donor single-excitation configuration
In some experiments, the receptor needs to be excited, for example, to confirm the distribution of all the receptor molecules, to photobleach the receptor molecules, and so on. This configuration consists of two excitation plates, one for the donor and one for the acceptor, and a dual-band dichroic mirror and two emitting plates. The two emitters need to be mounted on the filter runner, and the different emitters are switched through the rotor to receive the fluorescence of the donor and acceptor respectively. If the emitter is mounted in a beam splitter, an additional dichroic mirror for beam splitting is required.

Cy3 - Cy5 FRET pair,Donor and acceptor dual excitation configurations
 
Förster  Resonance Energy Transfer (FRET) is the transfer of photonic energy between two light-sensitive molecules (chromophores). When both chromophores are fluorophores, this is often referred to as Fluorescent Resonance Energy Transfer. The ‘donor’ molecule moves to an excited state after absorbing a photon.  
 
The ‘donor’ can then transfer this energy to the ‘acceptor’ chromophore via nonradiative dipole-dipole coupling. This energy transfer can only happen if the donor and acceptor are in close proximity to one another, making this technique very sensitive to small changes in chromophore separation.
 
To image FRET effectively, one must clearly quantify both the donor and the acceptor emission to ensure that any FRET signal does not contain direct fluorescence from either donor or acceptor chromophores.

 


ET Series - Widefield FRET Filter Set ( Donor and acceptor dual excitation)
Spectral Transmission Curves  
Model
 FRET Probe Target Excitation tablets
(Donor)		 Excitation tablets
(Receptor)		 Excite
Dichroic mirrors		 Transmitters
(Donor)		 Transmitters
( Receptor )		 Spectroscopy
Dichroic mirrors
89002  CFP/YFP ET430/24x ET500/20x 69008bs ET470/24m ET535/30m T505lpxr-UF
89021 GFP/RFP ET470/40x ET572/35x 59022bs ET525/50m ET632/60m T570lpxr-UF
89022 Cy3/Cy5 ET545/25x ET640/30x 59007bs ET595/50m ET690/50m T660lpxr-UF
 
 
ET Series - Surface Single Molecule FRET Filter Sets
Spectral Transmission Curves  
Model
Laser
 Excitation Tablets Excite
Dichroic mirrors		 Multi-band transmitters Transmitters
(Donor)		 Transmitters
( Receptor )		 Spectroscopy
Dichroic mirrors

 TRF59901-EM

 405/488 nm ZET405/488x ZT405/488rpc-UF2 ZET405/488m ET450/50m ET525/50m T495lpxr-UF

 TRF59902-EM

 442/514 nm ZET442/514x ZT442/514rpc-UF2 ZET442/514m ET480/40m ET545/40m T505lpxr-UF

 TRF59903-EM

 442/532 nm ZET442/532x ZT442/532rpc-UF2 ZET442/532m ET500/40m ET585/65m T525lpxr-UF

 TRF59904-EM

 488/561 nm ZET488/561x ZT488/561rpc-UF2 ZET488/561m ET525/50m ET610/75m T560lpxr-UF

 TRF59905-EM

 488/594 nm ZET488/594x ZT488/594rpc-UF2 ZET488/594m ET525/50m ET645/75m T570lpxr-UF

 TRF59906-EM

 488/640 nm ZET488/640x ZT488/640rpc-UF2 ZET488/640m ET525/50m ET655lp T570lpxr-UF

 TRF59907-EM

 532/640 nm ZET532/640x ZT532/640rpc-UF2 ZET532/640m ET585/65m ET655lp T635lpxr-UF

*Note Prices do not include any shipping, tax or customs clearance charges


Compared to conventional laser microscopes, TIRF has higher requirements for the flatness and thickness of dichroic mirrors. The flatness needs to be controlled at 0.5 waves/inch and below, and the thickness of the dichroic mirror must also be 2mm or more to avoid the influence of external stress on the flatness during installation.
 
Conventional microscope cubes cannot be fitted with dichroic mirrors with a thickness of 2 mm or more. In addition, because the material is mainly plastic, the resistance to deformation is also poor. For TIRF applications, Chroma has launched a patent-protected TIRF-specific cube (patent number: US8488238B2), which has the following features:
  • Ability to mount dichroic mirrors up to 3 mm thickness;
  • The ability to mount emitters with a maximum thickness of 6 mm allows the OD value to be increased in and near the laser line by superimposing two emitters;
  • The metal material has strong resistance to deformation and does not cause stress on the installed dichroic mirror.
Cube number Excitation tablets Dichroic mirrors Transmitters Microscope model  
91032 25 mm diameter 26 x 38 mm 25 mm diameter Nikon  TE2000/Ti
3.5 mm thickness（standard） 2mm thickness ( standard ) 6 mm thickness（maximum）
6 mm thickness（maximum） 3mm thickness ( maximum )
91041 25 mm diameter 26 x 38 mm 25 mm diameter Olympus  IX71/IX81
3.5 mm thickness（standard） 2mm thickness ( standard ) 6 mm thickness（maximum）
6 mm thickness（maximum） 3mm thickness ( maximum )
91044 25 mm diameter 26 x 38 mm 25 mm diameter Olympus  IX73/IX83
3.5 mm thickness（standard） 2mm thickness ( standard ) 6 mm thickness（maximum）
6 mm thickness（maximum） 3mm thickness ( maximum )
91042 25 mm diameter 25.5 x 36 mm 25 mm diameter Zeiss Axio
3.5 mm thickness（standard） 2mm thickness ( standard ) 6 mm thickness（maximum）
6 mm thickness（maximum） 3mm thickness ( maximum )
1,The Chroma TIRF cube is not sold separately and requires Chroma to install the TIRF dichroic mirror and correct its flatness;
2,TIRF dichroic mirrors are fixed with glue.

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Compare Model Drawings & Specs Availability Reference Price
(USD)
TRF59901-EM Filter Set
TRF59901-EM Filter Set, Laser 405/488 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59902-EM Filter Set
TRF59902-EM Filter Set, Laser 442/514 nm ，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59903-EM Filter Set
TRF59903-EM Filter Set, Laser 442/532 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59904-EM Filter Set
TRF59904-EM Filter Set, Laser 488/561 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59905-EM Filter Set
TRF59905-EM Filter Set, Laser 488/594 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59906-EM Filter Set
TRF59906-EM Filter Set, Laser 488/640 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
TRF59907-EM Filter Set
TRF59907-EM Filter Set, Laser 532/640 nm，Filters can be purchased separately 		4-6 Weeks $2100.00
89002 Filter Set
89002 ET Series - Sedat Multiband,Typical fluorophores CFP，YFP,Filters can be purchased separately 		4-6 Weeks $1600.00
89021 Filter set
89021 ET Series - Sedat Multiband,Typical fluorophores BFITC/GFP，mCherry,Filters can be purchased separately 		4-6 Weeks $1600.00
89007 Filter set
89007 ET Series - Sedat Multiband,Typical fluorophores CFP，YFP，Cy3/TRITC,Filters can be purchased separately 		4-6 Weeks $2200.00

91042 - Parameter

91044 - Parameter

91041 - Parameter

91032 - Parameter

89007 Filter set - Parameter

89021 Filter set - Parameter

89002 Filter Set - Parameter

TRF59906-EM Filter Set - Parameter

TRF59907-EM Filter Set - Parameter

TRF59905-EM Filter Set - Parameter

TRF59904-EM Filter Set - Parameter

TRF59903-EM Filter Set - Parameter

TRF59902-EM Filter Set - Parameter

TRF59901-EM Filter Set - Parameter

91042 - Download

91044 - Download

91041 - Download

91032 - Download

89007 Filter set - Download

89021 Filter set - Download

89002 Filter Set - Download

TRF59906-EM Filter Set - Download

TRF59907-EM Filter Set - Download

TRF59905-EM Filter Set - Download

TRF59904-EM Filter Set - Download

TRF59903-EM Filter Set - Download

TRF59902-EM Filter Set - Download

TRF59901-EM Filter Set - Download

Accessories

Compare Model Drawings & Specs Availability Reference Price
(USD)
91032
Laser TIRF for Nikon TE2000/Ti,Exciter (x) = 25mm, Emitter (m) = 25mm, Beamsplitter = 38x26mm 		4-6 Weeks Request for quote
91041
91041,Laser TIRF for Olympus BX2 Exciter (x) 25mm, Emitter (m) = 25mm, Beamsplitter = 26x36x2mm 		4-6 Weeks Request for quote
91044
91044,Laser TIRF for Olympus BX3/IX3 models, for 25mm filters,Exciter (x) = 25mm, Emitter (m) = 25mm, Beamsplitter = 26x36mm with a maximum thickness of 3mm 		4-6 Weeks Request for quote
91042
91042,Laser TIRF for Nikon TE2000/Ti,Exciter (x) = 25mm, Emitter (m) = 25mm, Beamsplitter = 38x26mm 		4-6 Weeks Request for quote

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