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LOCATION: Room 09.171.1 (BSL2)
SYSTEM SPECIFICATIONS:
- Imaging: Fast imaging with low phototoxicity
- Stand: Nikon Ti2 with perfect focus system (PFS)
- XY-stage: motorized
- Temperature and CO2 incubation
- Software: NIS elements with JOBS
- Yokogawa CSU-W1 confocal scan head (50um disk)
CAMERA SPECIFICATIONS:
- Model: 2x Hamamatsu ORCA-Fusion, sCMOS
- Effective number of pixels: 2304 x 2304
- Cell size: 6.5µm x 6.5µm
- Effective Area: 14.976mm x 14.976mm
- Max frame rate: 100 frames/second
- Dynamic range: 16 bit
- Quantum efficiency (peak): > 80% (@550nm)
- Read out noise: 0.7 - 1.4 electrons (depending on fps)
OBJECTIVES | ||||
Objective | Magnification / NA | Medium | Contrast | Working Distance |
CFI Plan Apo Lambda D | 4x / 0.2 | air | BF | 20 mm |
CFI Plan Apo Lambda D | 10x / 0.45 | air | BF | 4 mm |
CFI Plan Apo Lambda S | 25x / 1.05 | silicon | BF | 0.55 mm |
CFI Apo LWD Lambda S | 40x / 1.15 | water | BF | 0.6 mm |
| CFI Plan Apo Lambda D | 60x / 1.42 | oil | BF | 0.15 mm |
CFI Plan Apo Lambda | 100x / 1.45 | oil | BF | 0.13 mm |
ILLUMINATION lasers for spinning disk | ||
Wavelengths (nm) | Type | Some dyes and use |
405 | Diode laser | Alexa405, DAPI |
488 | Diode laser | Alexa488, GFP |
561 | Diode laser | Alexa568, RFP |
640 | Diode laser | Alexa647, Cy5 |
ILLUMINATION LED for widefield and eyepiece observation | ||
Wavelengths (nm) | Type | Some dyes and use |
| 385 | LED | Alexa405, DAPI |
475 | LED | Alexa488, GFP |
| 550 | LED | Alexa568, RFP |
621 | LED | Alexa647, Cy5 |
Main DICHROIC MIRROR | ||
Fluorochromes | Transmission to sample (nm) | Reflection to camera (nm) |
DAPI | > 80% 405 – 405 nm | > 95% 422 – 473 nm |
GFP, Alexa488 | > 80% 488 – 488 nm | > 95% 503 – 545 nm |
RFP, Alexa568 | > 80% 620 – 665 nm | > 95% 586 – 620 nm |
Cy5, Alexa647 | > 80% 638 – 647 nm | > 95% 665 – 750 nm |
Secondary DICHROIC MIRROR | |
Single camera mode | Full mirror to primary camera (back port) |
Dual camera mode | LP 561 nm beam splitter |
EMISSION FILTERS in front of the primary camera (back port) | |||
Fluorochromes | Availability | Bandpass | Transmission (nm) |
DAPI | YES | single BP | 417-477 |
GFP, Alexa488 | YES | single BP | 510-540 |
RFP, Alexa568 | YES | single BP | 574-626 |
Cy5, Alexa647 | YES | single BP | 671-745 |
EMISSION FILTERS in front of the secondary camera (left port) | |||
Fluorochromes | Availability | Bandpass | Transmission (nm) |
RFP, Alexa568 | YES | single BP | 574-626 |
Cy5, Alexa647 | YES | single BP | 671-745 |
Filter cubes (sets) for eyepiece observation | |||
Fluorochromes | Excitation filer (nm) | Dichroic cutoff (nm) | Emission filter (nm) |
DAPI | 377/50 | 409 | 447/60 |
GFP, Alexa488 | 466/40 | 495 | 525/50 |
RFP, Alexa568 | 562/40 | 593 | 641/75 |