Semrock Master Catalog 2018

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Fluorophores Single-band Sets Multiband Sets Cubes Laser Sets NLO Filters Individual Filters Dichroic Beamsplitters Tunable Filters Introduction to Fluorescence Filters Optical fluorescence occurs when a molecule absorbs light at wavelengths within its absorption band, and then nearly instantaneously emits light at longer wavelengths within its emission band. For analytical purposes, strongly fluorescing molecules known as fluorophores are specifically attached to biological molecules and other targets of interest to enable identification, quantification, and even real-time observation of biological and chemical activity. Fluorescence is widely used in biotechnology and analytical applications due to its extraordinary sensitivity, high specificity, and simplicity. Most fluorescence instruments, including fluorescence microscopes, are based on optical filters. A typical system has three basic filters: an excitation filter (or exciter), a dichroic beamsplitter (or dichromatic mirror), and an emission filter (or barrier filter). The exciter is typically a bandpass filter that passes only the wavelengths absorbed by the fluorophore, thus minimizing excitation of other sources of fluorescence and blocking excitation light in the fluorescence emission band. The dichroic is an edge filter used at an oblique angle of incidence (typically 45°) to efficiently reflect light Filter Transmission TECHNICAL NOTE excitation filter sample absorption Wavelength of Light dichroic beamsplitter emission filter sample fluorescence in the excitation band and to transmit light in the emission band. The emitter is typically a bandpass filter that passes only the wavelengths emitted by the fluorophore and blocks all undesired light outside this band – especially the excitation light. By blocking unwanted excitation energy (including UV and IR) or sample and system autofluorescence, optical filters ensure the darkest background. An appropriate combination of optical filters, making up a filter set, enables the visualization of a given fluorophore. See pages 10-13 for a listing of popular fluorophores and corresponding filter sets that can be used to image these fluorophores. A filter set needs to be optimized not only for imaging of distinct fluorophores but also designed to image a given fluorophore under different experimental conditions. Most of <strong>Semrock</strong> filter sets are a balance between high-brightness and high-contrast. These filter sets are the best choice of filters under standard imaging conditions. However, when the signal level from a sample is low, sets with wider passbands of the excitation and emission filters enable maximum signal collection efficiency. Studies such as imaging of single molecules typically utilize a filter set with a wide passband or a long pass emission filter. In studies utilizing such filter sets, it is required to maintain very low background autofluorescence signal by means of appropriate sample preparation protocols. However, since the wide passbands of such filter sets occupy a large spectral bandwidth, such filters are not preferred in multiplexing assays when imaging of several fluorophores is required. Filter sets with narrower passbands are preferred options when imaging a sample labeled with multiple fluorophores. Such filter sets reduce crosstalk between multiple fluorophores. Narrower passbands allow only the strongest portion of the fluorophore emission spectrum to be transmitted, reduce autofluorescence noise and thus improve the signal-to-noise ratio in high background autofluorescence samples. Such filter sets are ideal for samples with ample fluorescent signal level. In most fluorescence instruments, the best performance is obtained with thin-film interference filters, which are comprised of multiple alternating thin layers of transparent materials with different indices of refraction on a glass substrate. The complex layer structure determines the spectrum of light transmission by a filter. Thin-film filters are simple to use, inexpensive, and provide excellent optical performance: high transmission over an arbitrarily determined bandwidth, steep edges, and high blocking of undesired light over the widest possible wavelength range. FITC-5050A “High Brightness” Filter Set 100 90 80 70 145% 60 50 100% 71% 40 30 Brightness Contrast 20 10 0 400 450 500 550 600 650 Wavelength (nm) Advances in thin-film filter technology pioneered by <strong>Semrock</strong>, and embodied in all BrightLine filters, permit even higher performance while resolving the longevity and handling issues that can plague filters made with older soft-coating technology. This advanced technology is so flexible that users have a choice between the highest-performance BrightLine filter sets and the best-value BrightLine Basic filter sets. Transmission (%) Transmission (%) 100 90 80 70 60 50 40 30 20 10 FITC-2024B “High Contrast” Filter Set 100% 0 400 450 500 550 600 Wavelength (nm) 57% Brightness Contrast 650 140% More 14
BrightLine ® Single-band Sets Filter Set / Primary Fluorophores TRP-A Tryptophan Designed for UV fluorescence Use with UV LED or filtered Xe arc lamps, or detectors not sensitive to near-IR light. BUV395-3018A BD Horizon Brillant TM Ultraviolet 395 DAPI-11LP-A (Longpass) DAPI, Hoechst, AMCA, BFP, Alexa Fluor ® 350 DAPI-50LP-A (Longpass) DAPI, Hoechst, AMCA, BFP, Alexa Fluor ® 350 BV421-3824A BD Horizon Brillant TM Violet 421, DAPI, Alexa Fluor ® 405, DyLight405 DAPI-1160B (Highest Contrast) DAPI, Hoechst, AMCA, BFP, Alexa Fluor ® 350 DAPI-5060C (Highest Brightness) DAPI, Hoechst, AMCA, BFP, Alexa Fluor ® 350 BV480-2432A BD Horizon Brillant TM Violet 480, CFP, Cerulean, SpectrumAqua CFP-2432C CFP, AmCyan, SYTOX Blue, BOBO-1, Cerulean FURA2-C (Four Filter Set) 2+ Fura-2 Ca indicator, LysoSensor Yellow/Blue GFP-1828A (Highest Contrast) GFP, (EGFP), DiO, Cy2 , YOYO-1, YO-PRO-1 GFP-3035D (All Purpose) GFP, (EGFP), DiO, Cy2 , YOYO-1, YO-PRO-1 GFP-30LP-B (Longpass) GFP, (EGFP), DiO, Cy2 , YOYO-1, YO-PRO-1 GFP-4050B (Highest Brightness) GFP, (EGFP), DiO, Cy2 , YOYO-1, YO-PRO-1 FITC-2024B (Highest Contrast) FITC, rsGFP, Bodipy, 5-FAM, Fluo-4, Alexa Fluor ® 488 FITC-3540C (All Purpose) FITC, rsGFP, Bodipy, 5-FAM, Fluo-4, Alexa Fluor ® 488 FITC-5050A (Highest Brightness) FITC, rsGFP, Bodipy, 5-FAM, Fluo-4, Alexa Fluor ® 488 YFP-2427B YFP, Calcium Green-1, Eosin, Fluo-3, Rhodamine 123 CUBES PAGE 35 Excitation (CWL/BW) Emission (CWL/BW) Dichroic (Edge) Filter Set Part Numbers / –ZERO Set Part Numbers Base Price / –ZERO Price 280/20 357/44 310 nm TRP-A-000 $1025 356/30 392/18 376 nm BUV395-3018A BUV395-3018A-000-ZERO 387/11 409/LP 409 nm DAPI-11LP-A-000 DAPI-11LP-A-000-ZERO 377/50 409/LP 409 nm DAPI-50LP-A-000 DAPI-50LP-A-000-ZERO 389/38 433/24 414 nm BV421-3824A BV421-3824A-000-ZERO 387/11 447/60 409 nm DAPI-1160B-000 DAPI-1160B-000-ZERO 377/50 447/60 409 nm DAPI-5060C-000 DAPI-5060C-000-ZERO 438/24 483/32 458 nm BV480-2432A BV480-2432A-000-ZERO 438/24 483/32 458 nm CFP-2432C-000 CFP-2432C-000-ZERO Ex1: 340/26 Ex2: 387/11 510/84 409 nm FURA2-C-000 FURA2-C-000-ZERO 482/18 520/28 495 nm GFP-1828A-000 GFP-1828A-000-ZERO 472/30 520/35 495 nm GFP-3035D-000 GFP-3035D-000-ZERO 472/30 496/LP 495 nm GFP-30LP-B-000 GFP-30LP-B-000-ZERO 466/40 525/50 495 nm GFP-4050B-000 GFP-4050B-000-ZERO 485/20 524/24 506 nm FITC-2024B-000 FITC-2024B-000-ZERO 482/35 536/40 506 nm FITC-3540C-000 FITC-3540C-000-ZERO 475/50 540/50 506 nm FITC-5050A-000 FITC-5050A-000-ZERO 500/24 542/27 520 nm YFP-2427B-000 YFP-2427B-000-ZERO $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $1125 $1224 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 $785 $884 (continued) Filter Specifications on page 34 “–ZERO” denotes zero pixel shift performance (see page 36) Fluorophores Multiband Sets Cubes Laser Sets NLO Filters Dichroic Beamsplitters Tunable Filters Single-band Sets Individual Filters 15 More
Page 1 and 2: Semrock 2018 Master Catalog Everyth
Page 3 and 4: Welcome to the 2018 Master Catalog
Page 5 and 6: T T T T Laser & Optical System Filt
Page 7 and 8: SearchLight Optimization Calculator
Page 9 and 10: The Semrock Advantage Proven Reliab
Page 11 and 12: BrightLine ® Single-band Sets When
Page 13 and 14: BrightLine ® Single-band Sets for
Page 15: BrightLine ® Multiband Sets for Po
Page 19 and 20: TECHNICAL NOTE Ultraviolet (UV) Flu
Page 21 and 22: BrightLine ® Single-band Sets for
Page 23 and 24: Qdot ® Single-band Filter Sets Fil
Page 25 and 26: BrightLine ® FRET Single-band Sets
Page 27 and 28: BrightLine ® Basic TM Best-value S
Page 29 and 30: (continued from previous page) Fluo
Page 31 and 32: BrightLine ® Multiband LED Filter
Page 33 and 34: BrightLine ® Multiband Fluorescenc
Page 35 and 36: BrightLine ® Multiband Fluorescenc
Page 37 and 38: Fluorescence Filter Cubes See onlin
Page 39 and 40: BrightLine ® Laser Filter and Set
Page 41 and 42: BrightLine ® Laser Fluorescence Mu
Page 43 and 44: BrightLine ® Laser Fluorescence Fi
Page 45 and 46: TECHNICAL NOTE Sputtered Thin-film
Page 47 and 48: BrightLine ® Multiphoton LaserMUX
Page 49 and 50: Multiphoton Filters Common Specific
Page 51 and 52: BrightLine ® Single-band Bandpass
Page 59 and 60: BrightLine ® Long / Short pass Sin
Page 61 and 62: BrightLine ® Multiband Bandpass Fi
Page 63 and 64: BrightLine ® Multiband Bandpass Fi
Page 65 and 66: BrightLine ® Single-edge Dichroic
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BrightLine ® Multiedge Dichroic Be
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BrightLine ® Dichroic Beamsplitter
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Single-band Sets Fluorophores (cont
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BrightLine ® Super-resolution / TI
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BrightLine ® Laser Multiedge Dichr
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LaserMUX TM Beam Combining Filters
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TECHNICAL NOTE Measurement of Optic
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VersaChrome ® Tunable Filters TECH
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VersaChrome ® Tunable Bandpass Fil
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VersaChrome Edge TM Tunable Filters
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Laser Wavelength Reference Table La
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Polarization Filters Common Specifi
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General Purpose Mirrors Semrock gen
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EdgeBasic TM Long / Short Wave Pass
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Edge Steepness and Transition Width
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RazorEdge ® Common Specifications
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Filter Types for Raman Spectroscopy
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MaxLine ® Laser-line Spectra and S
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Near Infrared Bandpass Filters Tran
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StopLine ® Single-notch Filter Com
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TECHNICAL NOTE Filter Spectra at No
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Working with Optical Density Optica
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LEARN MORE WITH OUR VIDEO COLLECTIO
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The Physics of Pixel Shift Pixel sh
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Get the Right Solution, Right Now W
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