| Fluorescence Dye | Excitation filter (nm) | Emission filter (nm) | Dichroic mirror (nm) |
|---|---|---|---|
| Atto 425 | BP430/24 | BP480/30 | DM450 |
| BP430/24 | BP480/20 | DM450 | |
| FAM | BP470/30 | BP520/20 | DM505 |
| BP470/30 | BP525/20 | DM505 | |
| BP480/20 | BP520/20 | DM505 | |
| BP485/20 | BP525/20 | DM505 | |
| HEX,VIC,JOE | BP520/20 | BP565/22 | DM550 |
| BP525/20 | BP565/22 | DM550 | |
| BP530/20 | BP565/22 | DM550 | |
| BP530/20 | BP570/20 | DM550 | |
| CY3,TAMRA | BP543/20 | BP580/20 | DM565 |
| BP543/20 | BP585/20 | DM565 | |
| ROX | BP565/22 | BP615/40 | DM590 |
| BP570/20 | BP612/20 | DM590 | |
| BP580/20 | BP620/20 | DM600 | |
| BP585/20 | BP620/20 | DM600 | |
| Atto 633 | BP620/20 | BP655/20 | DM635 |
| CY5 | BP624/40 | BP694/40 | DM647 |
| BP624/40 | BP675/20 | DM647 | |
| BP630/20 | BP675/20 | DM647 | |
| BP630/20 | BP680/20 | DM647 | |
| CY5.5 | BP675/20 | BP725/30 | DM700 |
| BP665/22 | BP725/40 | DM700 | |
| CY7 | BP715/70 | BP800/60 | DM770 |
| BP725/30 | BP800/30 | DM770 | |
| BP725/40 | BP800/30 | DM770 |
1. Top Cy5 Alternatives in Fluorescence Imaging
Cy5 is widely used in far-red fluorescence imaging, but researchers often seek alternatives for specific needs. Two leading options stand out: - Alexa Fluor 647: Matches Cy5’s spectral profile with excitation ~650nm and emission ~668nm, offering superior photostability—ideal for long-term live-cell imaging. - Allophycocyanin (APC): A phycobiliprotein with excitation ~650nm and emission ~660nm, prized for high brightness in flow cytometry. Both alternatives share Cy5’s compatibility with red-channel imaging systems, making them drop-in replacements with minimal setup adjustments.
2. OPTOStokes Filters for Cy5 Substitutes
To maximize performance with these alternatives, filter sets must align with their spectral peaks. OPTOStokes provides: - Stock Filter Sets: Pre-calibrated for Alexa Fluor 647 (excitation 650/20nm, emission 670/20nm) and APC (excitation 650/15nm, emission 660/20nm). - Custom Solutions: Tailored bandwidths or dichroic mirrors (e.g., DM655nm) for unique imaging systems, ensuring zero crosstalk. All filters maintain >90% transmission in passbands, reducing exposure time—critical for photostability.
3. How to Prevent Photobleaching in Cy5 & Alternatives
Photobleaching (fluorescence loss from light exposure) undermines data reliability. Key strategies:
- Minimize Exposure: Use high-transmission filters (like OPTOStokes’ >90% models) to reduce laser power/imaging time.
- Anti-Fading Agents: Pair with reagents (e.g., DABCO) to slow dye degradation—compatible with all Cy5 substitutes.
- Filter Quality: Choose filters with tight blocking (OD6+ in off-bands) to avoid stray light accelerating bleaching.
- Regular Checks: Inspect filter surfaces for degradation; replace with OPTOStokes’ durable coatings (scratch-resistant) when needed.
4. Get Started with OPTOStokes
Need help with Cy5 alternatives or photobleaching?
- Email sales@optofilters.com for:
- Free spectral compatibility checks for your imaging system
- Custom filter designs for Alexa Fluor 647/APC
- Visit our site to: - Browse in-stock Cy5 substitute filter sets
- Request a no-obligation quote OPTOStokes: Precision filters for stable, high-quality fluorescence imaging—when you need them.
