Usage
- Contact CMN staff to be trained on this equipment or if you have questions about using this microscope.
- Online Calendar
MICROSCOPE PLATFORM (Nikon TiE Inverted, 2010):
Fully motorized (X-Y) stage with microwell plate and slide holders
Piezo stage (Z) available for resonance Z scanning
Perfect Focus infrared imaging system https://www.microscopyu.com/applications/live-cell-imaging/nikon-perfect-focus-system
1.5 x manual zoom
Microscopy
Infinity Corrected Lenses:
| Objective Magnification | Perfect Focus compatible | Numerical Aperature (NA) | Immersion Media | Working Distance (mm) | Objective Type | Primary Use |
| 10x | 0.45 | NA-Air | 4 | Plan Apo | Fixed Samples | |
| 20x | PF | 0.75 | NA-Air | 1 | Plan Apo | Fixed or Live |
| 20x | 0.75 | Multi (water, oil, glycerin) | 0.35 | Plan Fluor | Live Samples | |
| 60x | 1.2 | Water | 0.27 (0.13-0.19 mm) | Plan Apo | Live Samples | |
| 60x | PF | 1.4 | Oil | 0.13 | Plan Apo | Fixed Samples |
| 100x | 1.4 | Oil | 0.17 | Plan Apo | Fixed Samples |
Epifluorescent filters for sample visualization (no camera)
| Filter Set | Excitation | Dichroic | Emission (bandpass) |
| DAPI HC HiSN Zero Shift | 325-375 nm | 400 | 435-485 |
| GFP HC HiSN Zero Shift | 450-490 | 495 | 500-550 |
| TxRED HC HiSN Zero Shift | 540-580 | 585 | 593-668 |
CONFOCAL PLATFORM (Nikon A1R-HD, 2018):
4 diode Lasers (405 nm, 488nm, 561 nm, 640 nm)
4 PMTS (Standard: Blue and Far Red, GaASP: Green and Red)
32 channel spectral detector
Transmitted light Detector for Differential Interference Contrast (DIC) microscopy
NIS Elements Advanced software v 5.2
Galvonometric scanner: Max Pixel size 4096×4096, 512×512 scan speed 4 fps, zoom: 1-1000 variable
Resonant scanner: max. 1024×1024 pixels, 30 fps (512 x 512 pixels) to 230 fps
(512 x 64 pixels), 15,600 lines/sec (line speed). Zoom: 7 steps (1x, 1.5x, 2x, 3x, 4x, 6x, 8x)
Pinhole: Variable
Spectral Detector: image acquisition speed: 4 fps (256×256), 1000 lps; 2.5nm, 6nm or 10 nm wavelength range
Z step 0.025 um
Lasers and recommended fluorophores:
| channel | Laser | Color (pseudocolor) | PMT/Detector (virtual filter, BP nm) | Recommended Fluorophores (AF=alexafluor) |
| 1 | 405 Diode
100 mw Coherent |
Blue (blue) | 1 (450/50 480DCXR) | DAPI, Hoechst, AF 405, Pacific Blue, Cascade Blue, Coumarin |
| 2 | 488 Diode
|
Green (green) | 2 (485/40 520 DCXR
|
488: AF488, FITC, eGFP, Cy2, YO-Pro
|
| 3 | 561 Diode
25 mw Coherent |
Red (red) | 3 (600/50 640 DXCR) | AFs 546, 551, 568, Cy3, DiI, DsRed, mCherry, PE, PI, RFP, tdTomato, TRITC |
| 4 | 640 Diode
100 mw Coherent |
Far Red (pink) | 4 (650 LP 3FW) | AF 633, 647, Cy5, To-PRO-3 |