Widefield (epifluorescence)
DANTE - Olympus IX81 widefield (Porter B047A)
Light Sources:
- Excelitas X-Cite Xylis 365nm, model XT720S. The Xylis. Super high-intensity capable LED, intensity selectable in 1% increments, 5-100%. Excitation bandwidth from UV (365nm) to Near-IR (770nm) with peaks at 365, 430, 475, 545, 650, and 735nm, matching optimally with common fluorophores.
- Halogen for brightfield and phase-contrast.
Standard filter configurations
This system utilizes the finest optical filters from Semrock and Chroma and employs separate excitation and emission wheels for maximum flexibility and customizable optical settings. This system employs fluor-matched single dichroic beamsplitters, and GFP/RFP dual and DAPI/FITC/TRITC/Cy5/Cy7 penta beam splitters; the multi-and beam splitters allow more rapid multi-channel imaging. These components together provide maximum brightness and fast switching between wavelengths to provide the best results.
Excitation filters:
- 325-375 (nm) (Chroma 49000); DAPI, Hoechst, Alexa350, 405, etc.
- 426-450 (nm) (Semrock); CFP, cerulean, etc.
- 450-490 (nm) (Chroma 49002); Alexa488, FITC, EGFP, etc.
- 459-481 (nm) (Semrock Brightline GFP/DsRed- 2X2M-C-000); Alexa488, FITC, EGFP, etc.
- 483-512 (nm) (Omega XF104-2); YFP, Alexa532, Venus, etc.
- 560-580(nm) (Chroma 41034); Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, etc.
- 546-566 (nm) (Semrock Brightline GFP/DsRed- 2X2M-C-000); Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, etc.
- 618-648 (nm) (Semrock Brightline 4040A); Alexa647, Cy5, etc.
- 672.5-747.5 (nm) (Chroma 49007); Alexa680/700/750, Cy7, etc.
Emission filters:
- 435-485 (nm) (Chroma 49000); DAPI, Hoechst, Alexa350, 405, etc.
- 472-494 (nm) (Semrock); CFP, cerulean, etc.
- 502.5-527.5 (Chroma 49002); Alexa488, FITC, EGFP, etc.
- 499-529 (nm) (Semrock Brightline GFP/DsRed- 2X2M-C-000); Alexa488, FITC, EGFP, etc.
- 527.5-562.5 (nm) (Omega XF104-2); YFP, Alexa532, Venus, etc.
- 590-650 (Chroma 41034); Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, etc.
- 580.5-653.5 (nm) (Semrock Brightline GFP/DsRed- 2X2M-C-000); Alexa555, 568, 594, TxRed, mRFP, mCherry, TdTomato, etc.
- 672-712 (nm) (Semrock Brightline 4040A); Alexa647, Cy5, etc.
- 765-855 (nm) (Chroma 49007); Alexa680/700/750, Cy7, etc.
Special Dichroic mirrors
- GFP/DsRed dual-edge dichroic (Semrock BrightLine GFP/DsRed- 2X2M-C-000)
- DAPI/GFP/RFP/CY5/Cy7 penta-edge dichroic (Semrock BrightLine FF409/493/573/652/759-Di01 25x36)
Other imaging
- Brightfield
- Phase-contrast (PH2 20x and 40x objectives)
Standard objectives:
- 4x 0.13NA UPlanFl N UIS2 WD 17 (mm)
- 10x 0.40NA UPlanApo Ph1 WD 3.1 (mm)
- 20x 0.70NA UPlanApo Ph2 WD 0.65 (mm)
- 40x 0.75NA UPlanFl N Ph2 WD 0.51 (mm)
- 60x 1.35NA UPlanSapo OIL WD 0.15 (mm)
- 100x 1.40NA UPlanSapo OIL WD 0.13 (mm)
Available upon request:
- 60X 1.20NA UPLSAPO WATER WD 0.28 (mm)
- Inquire
- Hamamatsu Orca R2 CCD. 12/16 bit
- 1344 x 1024, 6.45 x 6.45 micron pixels
- binning, 2x2, 4x4, 8x8
- peak QE >70%
- wavelength range >30% QE, 360 - 800nm (low light mode)
- High-speed large area montaging via Prior Scientific H117E2IX motorized stage with linear encoders, with advanced stitching in up to 7 wavelengths (inquire). Slide scanning for up to 4 standard 25x75mm slides per run; entirety of slide(s), multiple areas across all slides, via Prior stage insert. Includes pre-defined settings for 1, 2, 3, or 4 slides in diffferent orientations. Pre-defined slide scanning or large 50x75mm slide format, via Prior stage insert.
- Stage and well-plate navigator interface with pre-programmed plate formats for 6, 12, 24, 48, 96 wells and more. Advanced 2D and 3D deconvolution; Experiment design GUI; Debluring; Fluorescence unmixing; Image analysis; High dynamic range imaging (HDRI); more!!
- Save your own custom settings including optical configurations, scan areas, acquisition settings, for routine precision.
- Live cell imaging with InVivo Sci. stage-top humidified chamber, STEV.ECU.HC5; 1x35mm, multi-well plates, pre-mixed 5% CO2, balance air (please contact Dr. Orth); Oustanding phase contrast (20x and 40x); precise repeatability over time via Prior Scientific H117E2IX motorized stage with linear encoders; Z-drive; FRET (CFP --> YFP); GFP/mCherry dual dichroic or Blue/Green/Red/FarRed/Near-IR penta beam splitters provide for rapid multicolor live sample imaging.
Contact Director, James.Orth@olorado.edu
PEPPER - Nikon TiU widefield with RGB color brightfield (Porter B047A)
Light Sources:
- X - Cite 120
- Halogen illumination for brightfield and DIC
Standard filter configurations:
Semrock BrightLine quad Sedat
- DAPI, Hoechst, etc.
- Alexa488, FITC, GFP, etc.
- TRITC, Alexa555, 568, 594, mRFP, mCherry, TdTomato, etc.
- Alexa647, Cy5, etc.
Standard objectives:
- 4x 0.13NA Plan Fluor WD 17.2 (mm)
- 10x 0.45NA Plan Apo DIC N1 WD 4 (mm)
- 20x 0.50NA Plan Fluor WD 2.1 (mm)
- 40x 0.60NA Plan Fluor WD 1.8 (mm)
- 100x 1.30NA Plan Fluor Ph3 OIL WD 0.20 (mm)
Optional objectives:
- 2x 0.10NA Plan Apo WD 8.5 (mm)
- More (please inquire)
- CoolSNAP HQ2 monochrome camera for fluorescence
- Nikon DS-Fi3 24 bit high definition color camera for color brightfield (e.g. eosin, hematoxylin, crystal violet. etc.). This camera uses a CMOS chip with 5.9 megapixels.
ASI MS-2000 XYZ motorized stage. Sample holders for standard slide, 35 and 60 mm dishes, and multiwell plates. Brightfield color RGB imaging. Multi-point imaging. Montaging with stitching. Live cell imaging with TOKAI-HIT Cell Incubator; 1 X 35 (mm) dish or chamber slide (coverslip bottom). Rapid 4 color imaging via quad Sedat filter set (Semrock BrightLine).
