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BrightLine® single-band filter set, optimized for FITC and other like fluorophores

Semrock Set / Fluorophore Graphing

Part Number: FITC-3540C-000

Fluorophore Displayed:
Please select your fluorophore from the Fluorophore Tab below
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Semrock Filter Set: FITC-3540C

BrightLine® single-band filter set, optimized for FITC and other like fluorophores

This set is optimized for the most popular green fluorophores like FITC (fluorescein isothiocyanate), 5-FAM, and Alexa Fluor 488. It provides the both high brightness and signal-to-noise ratio. It is also ideal for similar fluorophores that have narrower, more closely spaced excitation and emission peaks, like BODIPY.

Individual filters and filter sets (no cube):
These filters sets contain individual filters. For filter sets already mounted in a holder/cube, scroll down to see complete mounted sets with cubes.
Part NumberPrice1Stock StatusQuantity
Click to expand/collapse details FITC-3540C-000 (Standard size set, fits most microscopes2)$825Confirmed AROAdd To Cart
Filter RoleFilter SizePart NumberPrice1
Single Band Exciter25 mm x 5.0 mmFF01-482/35-25$345
Single Band Emitter25 mm x 3.5 mmFF01-536/40-25$345
Single Band Dichroic25.2 mm x 35.6 mm x 1.1 mmFF506-Di03-25x36$285
Click to expand/collapse details FITC-3540C-000-ZERO (Standard size -ZERO pixel set)$924Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-L01-ZERO (Leica 'Large' -ZERO pixel set)$924Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-U02 (Olympus large-size (FX) set for U-M710I)$1,370Confirmed AROAdd To Cart

1) US domestic pricing only. If you are ordering from outside the US, please contact your nearest regional distributor for the correct list price.
2) Customers installing filters into their own Nikon Quadfluor cubes should contact us before ordering a filter set.

Complete filter sets mounted in microscope holders/cubes:
These sets are delivered already mounted in microscope filter holders, often called 'cubes', ready to be dropped into your microscope.
Nikon Quadfluor for microscopes: E200, E400, E600, E800, E1000, TS100, TS100F, TE200, TE300, ME600L, L150A, and some Labophot, Optiphot, and Diaphot series
Click to expand/collapse details FITC-3540C-NQF (set mounted in cube)$1,220Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-NQF-ZERO (-ZERO pixel set mounted in cube)$1,319Confirmed AROAdd To Cart
Nikon TE 2000 for microscopes: TE 2000, 50i, 55i, 80i, 90i, Eclipse Ti, Ni, and Ci series, and any using the Epi-fluor Illuminator
Click to expand/collapse details FITC-3540C-NTE (set mounted in cube)$1,220Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-NTE-ZERO (-ZERO pixel set mounted in cube)$1,319Confirmed AROAdd To Cart
Olympus U-MF2 for microscopes: AX70, BX, BX41, BX50, BX51, BX60, BX61, BX50/51WI, BX60/61WI, IX50, IX51, IX70, IX71, IX81
Click to expand/collapse details FITC-3540C-OMF (set mounted in cube)$1,260Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-OMF-ZERO (-ZERO pixel set mounted in cube)$1,359Confirmed AROAdd To Cart
Olympus U-FF for microscopes: BX53, BX63
Click to expand/collapse details FITC-3540C-OFF (set mounted in cube)$1,260Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-OFF-ZERO (-ZERO pixel set mounted in cube)$1,359Confirmed AROAdd To Cart
Olympus U-M710I for microscopes: Requires a 32mm emitter & exciter and 32x44mm dichroic beamsplitter. For IX53, IX73, & IX83 model microscopes
Click to expand/collapse details FITC-3540C-OFX (set mounted in cube)$2,065Confirmed AROAdd To Cart
Zeiss FL Cube EC P+C for microscopes: Axio Imager, Axiostar Plus, Axioskop 40, Axioskop 2 (post-2001), Axioplan 2i, Axioplan 2ie, Axiovert 200, Axiovert 40, Axio Observer, Axio Examiner and Axio Scope A1
Click to expand/collapse details FITC-3540C-ZHE (set mounted in cube)$1,100Confirmed AROAdd To Cart
Click to expand/collapse details FITC-3540C-ZHE-ZERO (-ZERO pixel set mounted in cube)$1,199Confirmed AROAdd To Cart
Leica DM-K for microscopes: DM-2000, DM-2500, DM-3000, DMI3000 B, DM-4000, DMI4000 B, DM-5000, DM-5500, DM-6000, DMI6000 B, DM4 and DM6
Click to expand/collapse details FITC-3540C-LDMK-ZERO (-ZERO pixel set mounted in cube)$1,274Confirmed AROAdd To Cart
Leica DMi8 (P-Cube) for microscopes: DMi8
Click to expand/collapse details FITC-3540C-LDMP-ZERO (-ZERO pixel set mounted in cube)$1,359Confirmed AROAdd To Cart
Aperio AMF for microscopes: ScanScope FL
Click to expand/collapse details FITC-3540C-AMF-ZERO (-ZERO pixel set mounted in cube)$1,389Confirmed AROAdd To Cart

1) US domestic pricing only. If you are ordering from outside the US, please contact your nearest regional distributor for the correct list price.
Optical Specifications
BlockingBrightLine filters have blocking far exceeding OD 6 as needed to ensure a black background, even when using modern low-noise CCD cameras. The blocking is optimized for microscopy applications using our exclusive SpecMaker™ fluorescence filter design.

General Filter Specifications
Microscope CompatibilityBrightLine filters are available to fit Leica, Nikon, Olympus, Zeiss, and Aperio microscopes.
Optical Damage RatingTesting has proven to show no signs of degradation when exposed to at least 6.0 W of power from an unfiltered xenon arc lamp over a 25 mm diameter (corresponding to 1.2 W/cm2) for over 500 hours.

Physical Filter Specifications
Exciter/Emitter OrientationArrow on ring indicates preferred direction of propagation of light.
Dichroic Orientation"Reflective coating side" should face toward light source and sample.

5-Star Optimized – Filter set is specifically optimized for that fluorophore.
3-Star Compatible – Should result in nearly ideal performance in most situations.
1-Star Acceptable – Actual performance is dependent on specific experimental conditions.
FluorophorePeak Excitation
Wavelength (nm)
Peak Emission
Wavelength (nm)
CompatibilityDisplay on
Plot above
5-FAM (5-carboxyfluorescein)4925183-Star
6-carboxyrhodamine 6G5255551-Star
Acridine Orange (+DNA)5025263-Star
Acridine Yellow4705501-Star
Alexa Fluor® 4304315411-Star
Alexa Fluor® 4884995205-Star
Alexa Fluor® 5005035253-Star
ATTO 4885005253-Star
ATTO 4954955273-Star
BCECF (pH 5.5)4815183-Star
BCECF (pH 9.0)5025283-Star
BODIPY FL5055123-Star
BODIPY FL-MeOH5025113-Star
Calcium Green-15075291-Star
CyQUANT GR-DNA5025233-Star
Dendra2 (Green)4915071-Star
ecliptic pHluorin pH5.54735071-Star
FITC (Fluorescein)4955195-Star
Fluorescein dextran5015243-Star
Fluorescein-pH 8.04895173-Star
FluoSpheres Yellow-Green fluorescent microspheres5035143-Star
FM 1-434735791-Star
GFP (EGFP)4895113-Star
Green 4964965203-Star
Green 5005015243-Star
HCS LipidTOX Green neutral lipid stain4985071-Star
HCS LipidTOX Green phospholipidosis5045363-Star
HiLyte Fluor™ 4884975263-Star
LIVE-DEAD Fixable Green Dead Cell Stain4985253-Star
LysoSensor Green4485021-Star
LysoTracker Green5015091-Star
LysoTracker Yellow HCK-1234885651-Star
Magnesium Green5075311-Star
MitoTracker™ Green4905125-Star
NBD-X (MeOH)4675383-Star
NeuroTrace 500/525 Green Fluorescent Nissl Stain4955243-Star
Oregon Green™ 4884985265-Star
Oregon Green™ 5145135321-Star
Qdot® 525 Nanocrystals3005251-Star
Qdot® 545 Nanocrystals3005431-Star
Rhodamine 1104975195-Star
Rhodamine 1235075291-Star
Rhodamine Green4975233-Star
Rhodol Green4975243-Star
sgGFP™ (super glow GFP)4725061-Star
Sodium Green5075321-Star
SYBR Gold nucleic acid gel stain-DNA4965393-Star
SYBR Green I nucleic acid gel stain-DNA4985223-Star
SYBR Safe DNA gel stain-DNA5095261-Star
SYTO 115065251-Star
SYTO 134485061-Star
SYTO 164895203-Star
SYTO 94835001-Star
SYTO RNASelect geen fluorescent cell stain5035273-Star
SYTOX Green-DNA5045243-Star
Vybrant DyeCycle Green5065343-Star
wtGFP (wild type GFP, non-UV excitation)4745091-Star

Tech Note
Cleaning Optical FiltersInstructions for cleaning hard coated optical filters
Cube Assembly InstructionsPDF and video instruction for installing filters into popular filter cubes
Filter ReliabilityCompares the performance and reliability of hard coated filters to traditional soft-coated filters and provides information about the testing standards for Semrock filters
Introduction to Fluorescence FiltersShows how optical filters are used in a traditional fluorescence microscope and describes important factors to consider when choosing optical filters.
Orientation of Filters in a MicroscopeA guide to properly orienting Semrock filters based on the filter markings.
What is Pixel Shift?Explains how filter wedge angle causes pixel shift and describesthe impact of pixel shift on image fidelity
Journal Article
Achieving the Best Alignment for Fluorescent ImagesTuran Erdogan, PhD, and Atul Pradhan, PhD, Semrock Inc., BioPhotonics, August 2005
Semrock's BrightLine ZERO filter sets eliminate the beam deviation that can cause pixel shift when combining single-color images to create a multicolor image
White Paper
Spectral Modeling in Fluorescence MicroscopyDiscusses various sources of noise in fluorescence microscopy and how to maximize signal-to-noise ratios using software, sample preparation and optical filter selection
High Performance MicroscopyAn overview of high performance microscopy, including solutions from Semrock and additional learning resources.

FITC-3540C FITC 506 536/40 482/35 Single-Band FF506-Di03 FF01-536/40 FF01-482/35 FF506 Filter Filters Set Sets 5-FAM 5-carboxyfluorescein Acridine-Orange-(+DNA) Alexa-Fluor-488 AF488 Alexa488 Alexa-Fluor-500 AF500 Alexa500 ATTO-488 ATTO-495 BCECF BODIPY-FL Calcein CFP2 CyQUANT-GR-DNA Dronpa DY-505-Phalloidin Emerald Fluorescein Fluo-3 Fluo-4 Fluoro-Emerald FluoSpheres-Yellow-Green GFP EGFP Green-496 Green-500 MitoTracker-Green mWasabi Oregon-Green-488 Rhodamine-110 FITC-3540C-000 FITC-3540C-000-ZERO FITC-3540C-L01-ZERO FITC-3540C-U02 FITC-3540C-NQF FITC-3540C-NQF-ZERO FITC-3540C-NTE FITC-3540C-NTE-ZERO FITC-3540C-OMF FITC-3540C-OMF-ZERO FITC-3540C-OFF FITC-3540C-OFF-ZERO FITC-3540C-OFX FITC-3540C-ZHE FITC-3540C-ZHE-ZERO FITC-3540C-LDMP-ZERO FITC-3540C-LDMK-ZERO FITC-3540C-AMF-ZERO

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