The SNOUTY is a single objective light sheet microscope, with a tilted plane that leaves the objective at a 30-degree angle, designed to capture cells and tissues by the volume at high-speed with low background and minimal photobleaching/toxicity. The microscope is equipped with a Hammamtsu Orca-Quest-2 camera and a 60x 1.3NA silicon lens (can be swapped out with a water lens for expanded samples), for a final pixel size of 102nm and a field of view that is 400um diameter. The system contains 4 laser lines (405/488/561/639nm), that can be used with a quadpass filter for speed or single pass filters for specificity. A separate 808 laser as also been implemented to help suppress the triplet state of green fluorescent proteins can be used with the green flrouescent proteins to help prevent bleaching (please note only works with green because of physics).
For live imaging the Stage is equipped with an Oko Labs stage top incubator, 5% CO2, and a Prior pure focus system.
This machine is hooked up directly to our server because of large data sets. Please email us about using our server before use!
Capabilities
- 4-color fluorescence imaging
- Fast 3D imaging- automatically collect in volumes!
- Automated imaging
- Live cell time lapse imaging
- High speed imaging
- 808nm triplet depletion laser for Green fluorescent proteins
- Light sheet- minimal photo bleaching and low noise
Objectives:
- Plan Apo λ 60x/1.3NA Sil 0.3 WD
- Plan Apo 60x / 1.2 WI (can be switched out)
Equipment Details:
- Homebuit by Tanner Fadero
- Inverted stage
- Oko Stage Top Incubator with CO2
- Adapters for 96 Well plate, standard slide (25mm x 75mm), 35mm Dish, and Lab-Tek II Chambered Coverslips
- See slide and dish recommendations here: Plates and dishes for imaging, Note Must be No. 1.5 Coverslip equivialant
- Vortran VersaLase (8) system For Light Sheet
- ND filter wheel for Laser intensity control
- Vortrain Triplet Depletion Laser
- Prior Pure Focus system
- NanoScan SP400 Piezo Sample Scanner with 400 micron travel range (in process of being set up)
- Carin Opto spin filter wheels
Dithering mirror for light sheet homoginization
Excitation lasers for Light Sheet:
- VersaLase (8) system For Light Sheet
- 405nm, 100mW
- 488nm, 200mW
- 561nm, 50mW
- 639nm, 200mW
- Triplet Depletion Laser
- Stradus 808nm, 250mW
- NOTE: only works for green fluorophores
Brightfield:
- Dual LED for Differential phase contrast
Emission Filters:
- ZET405/488/561/640mv2 (Chroma)
- ZET488/561m (Chroma)
- ET445/58m (Chroma)
- ET535/70m (Chroma)
- ET630/75m (Chroma)
- ET706/95m (Chroma)
- ET750sp-2p8 (Chroma)
ND filters for additional laser power control:
- ND Filter OD 3.0 (0.1% of the light through)
- ND Filter OD 2.0 (1.0% of the light through)
- ND Filter OD 1.5 (3.16% of the light through)
- ND Filter OD 1.0 (10% of the light through)
- ND Filter OD 0.5 (31.6% of the light through)
- ND Filter OD 0.0 (100% of the light through)